Fani Koukouli

Marie Rooy

Dimitrios Tziotis

Kurt A Sailor

Heidi C O'neill

Josien Levenga

Mirko Witte

Michael Nilges

Jean-Pierre Changeux

Charles A Hoeffer

Jerry A Stitzel

Boris S Gutkin

David A Digregorio

Uwe Maskos
email: umaskos@pasteur.fr

Dphil

Nicotine reverses hypofrontality in animal models of addiction and schizophrenia

 . Recently, genome-wide association studies (GWAS) identified single-nucleotide polymorphisms (SNPs) in the human CHRNA5 gene, encoding the α5 nAChR subunit, that increase the risks for both smoking and schizophrenia 7,8 . Mice with altered nAChR gene function exhibit PFC-dependent behavioral deficits 9-11 , but it is unknown how the corresponding human polymorphisms alter the cellular and circuit mechanisms underlying behavior. Here we show that mice expressing a human α5SNP exhibit neurocognitive behavioral deficits in social interaction and sensorimotor gating tasks. Two-photon calcium imaging in awake mouse models showed that nicotine can differentially influence PFC pyramidal cell activity by nAChR modulation of layer II/III hierarchical inhibitory circuits. In α5-SNP-expressing and α5-knockout mice, lower activity of vasoactive intestinal polypeptide (VIP) interneurons resulted in an increased somatostatin (SOM) interneuron inhibitory drive over layer II/III pyramidal neurons. The decreased activity observed in α5-SNP-expressing mice resembles the hypofrontality observed in patients with psychiatric disorders, including schizophrenia and addiction 5,12 . Chronic nicotine administration reversed this hypofrontality, suggesting that administration of nicotine may represent a therapeutic strategy for the treatment of schizophrenia, and a physiological basis for the tendency of patients with schizophrenia to self-medicate by smoking 13 .

Impairment of cognitive processes that are critically reliant on the PFC is a signature of schizophrenia [START_REF] Barch | Selective deficits in prefrontal cortex function in medication-naive patients with schizophrenia[END_REF] and is associated with low activity of the default mode network [START_REF] Raichle | The Brain's Default Mode Network[END_REF] . However, the cellular and circuit deficits underlying 'hypofrontality' remain unexplored. Because acetylcholine (ACh) release results in strong modulation of cortical activity via nAChRs [START_REF] Robbins | Differential regulation of fronto-executive function by the monoamines and acetylcholine[END_REF][START_REF] Poorthuis | Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors[END_REF][START_REF] Sarter | Phasic acetylcholine release and the volume transmission hypothesis: time to move on[END_REF] , it is not surprising that human nAChR gene variants are strongly linked to mental disorders with cognitive deficits [START_REF] Sinkus | The human CHRNA7 and CHRFAM7A genes: A review of the genetics, regulation, and function[END_REF] . Genetic variants within the CHRNA3-CHRNA5-CHRNB4 nAChR gene cluster contribute to the risk of habitual smoking [START_REF]Genome-wide meta-analyses identify multiple loci associated with smoking behavior[END_REF] . Among the series of polymorphisms composing this human haplotype, the rs16969968 SNP leads to the substitution of aspartic acid at residue 398 by asparagine (D398N) in the human α5 subunit [START_REF] Bierut | Variants in nicotinic receptors and risk for nicotine dependence[END_REF] . Both in vitro and ex vivo studies indicate that this SNP results in partial loss of function of α5-containing nAChRs [START_REF] Bierut | Variants in nicotinic receptors and risk for nicotine dependence[END_REF][START_REF] Sciaccaluga | Crucial role of nicotinic 5 subunit variants for Ca2+ fluxes in ventral midbrain neurons[END_REF] . In humans, this SNP is associated with reduced resting-state functional connectivity, and circuits involving the PFC are altered [START_REF] Hong | A genetically modulated, intrinsic cingulate circuit supports human nicotine addiction[END_REF] .

Recently, the same genetic locus was associated with schizophrenia in a major GWAS [START_REF] Ripke | Biological insights from 108 schizophrenia-associated genetic loci[END_REF] , but it

is not yet clear whether this SNP is responsible for the hypofrontality observed in individuals with schizophrenia, which would directly implicate altered nicotinic signaling in this pathophysiology of schizophrenia. One obstacle preventing further mechanistic dissection of schizophrenia is the absence of animal models. Because genetic deletion of the α5 nicotinic subunit in mice leads to PFC-linked behavioral deficits, altered cholinergic excitation and aberrant neuronal morphology in the PFC [START_REF] Proulx | Nicotinic acetylcholine receptors in attention circuitry: the role of layer VI neurons of prefrontal cortex[END_REF] , we developed a mouse line expressing the human α5 variant to investigate the impact of this variant on behaviors associated with schizophrenia and to determine the neuronal and circuitry mechanisms underlying cognitive deficits.

Research on social cognition suggests that social interaction deficits, similar to emotional dysfunctions, represent characteristic markers of psychiatric disorders [START_REF] Derntl | Deficits in social cognition: a marker for psychiatric disorders?[END_REF] . Because impaired social ability is a common feature of schizophrenia [START_REF] Derntl | Deficits in social cognition: a marker for psychiatric disorders?[END_REF] , we used the three-chamber social test [START_REF] Nadler | Automated apparatus for quantitation of social approach behaviors in mice[END_REF] to determine whether α5-SNP-expressing (α5SNP) mice exhibit impairments in social interaction. The results indicated that α5SNP mice have deficits in sociability. Wild-type (WT) mice exhibited a clear preference for a novel social partner, spending 69.6 ± 2.9% of their time engaged in social interaction, while α5SNP littermates showed no preference between the social partner and an inanimate object (44.1 ± 5.3%) (Fig. 1a). The differences in social interaction cannot be attributed to the effects of differing genotypes on activity or general exploratory behavior because the WT and α5SNP mice did not differ in locomotor activity or combined time in the social and inanimate object interaction areas (Fig. 1a and Supplementary Discussion).

Deficiencies in the prepulse inhibition (PPI) task are a measure of impaired sensorimotor gating and are observed in several human neuropsychiatric disorders [START_REF] Braff | Human studies of prepulse inhibition of startle: normal subjects, patient groups, and pharmacological studies[END_REF] . The PPI task measures the ability of an animal to inhibit its startle in response to an acoustic tone (pulse) when the tone is preceded by a lower-intensity stimulus (prepulse). Examination of PPI has been particularly important in studying the neurobiology of schizophrenia because deficits in cognitive information gating are a clinically important feature of the disorder [START_REF] Braff | Human studies of prepulse inhibition of startle: normal subjects, patient groups, and pharmacological studies[END_REF] . There is a high degree of similarity between measures of PPI in rodent models and humans, which is in accordance with the use of PPI as a cross-species measure of sensorimotor gating [START_REF] Braff | Human studies of prepulse inhibition of startle: normal subjects, patient groups, and pharmacological studies[END_REF] . We found that α5SNP mice exhibited decreased PPI across a range of prepulse intensities (4, 8, 12 and 20 dB above background noise) relative to their WT littermates (Fig. 1b and Supplementary Discussion), suggesting that sensorimotor gating is disrupted by the α5 variant.

To determine the circuit mechanisms underlying the cognitive deficits attributed to dysfunction of α5-containing nAChRs and the human SNP, we assessed resting-state network activity using in vivo two-photon calcium imaging in the prelimbic cortex (PrLC) of the PFC in awake mice either lacking the α5 subunit (α5KO mice) or with knock-in of the α5 SNP (α5SNP mice) in comparison to WT mice. Neurons were transduced with an adeno-associated viral vector (AAV) expressing the genetically encoded calcium indicator GCaMP6f in order to monitor Ca 2+ transients as a proxy for action potential firing (Fig. 1c and Supplementary Fig. 1). Because we estimate that 80% of the labeled neurons in layers II/III are pyramidal neurons (Supplementary Fig. 2), we consider that such experiments primarily indicate the activity of the pyramidal neuron population. Interestingly, we found that the median frequency and duration of Ca 2+ transients were lower in both strains of transgenic mice than in WT mice (Supplementary Fig. 3). Spontaneous Ca 2+ transients were deconvolved with a putative unitary (action potential-evoked) fluorescence response in order to estimate neuronal firing rates (Supplementary Figs. 456and Supplementary Table 1). We found significantly lower layer II/III neuronal activity in the PrLC of α5KO (8.2 ± 0.5 spikes/min) and α5SNP (20.3 ± 0.6 spikes/min) mice when compared to WT mice (27.5 ± 0.9 spikes/min; Fig. 1d-f).

Thus, loss of function of α5-containing nAChRs causes a decrease in layer II/III microcircuit activity, which parallels the decrease in hypofrontality that is a defining feature of schizophrenia in humans [START_REF] Barch | Selective deficits in prefrontal cortex function in medication-naive patients with schizophrenia[END_REF] .

The α5 subunit is present only in VIP interneurons in layers II/III [START_REF] Porter | Selective excitation of subtypes of neocortical interneurons by nicotinic receptors[END_REF] , which exert inhibitory control of SOM and parvalbumin (PV) interneurons in the PFC [START_REF] Pi | Cortical interneurons that specialize in disinhibitory control[END_REF] . Since layer II/III pyramidal neurons do not express nAChRs, we hypothesized that loss of function of the α5 subunit would result in disinhibition of PV and SOM interneurons and, thus, reduced pyramidal cell activity (Fig. 1g). Because dendritic-targeting SOM interneurons express different nicotinic receptor subtypes, namely α7-and β2-containing nicotinic receptors [START_REF] Poorthuis | Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors[END_REF][START_REF] Poorthuis | Layer-specific interference with cholinergic signaling in the prefrontal cortex by smoking concentrations of nicotine[END_REF] , from perisomatictargeting PV cells [START_REF] Poorthuis | Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors[END_REF][START_REF] Kawaguchi | GABAergic cell subtypes and their synaptic connections in rat frontal cortex[END_REF] , which exclusively express α7-containing nAChRs, we compared PFC activity across transgenic mice with specific deletions of nAChR subunits differentially expressed in PV and SOM interneurons. We observed a strong increase in the median spike frequency of GCaMP6f-expressing pyramidal neurons in both KO lines, with the β2KO line exhibiting the highest frequencies (85 ± 1.1 spikes/min) in comparison to the α7KO line (45.9 ± 1.8 spikes/min) and WT mice (27.5 ± 0.9 spikes/min; Fig. 1h-j). Thus, the absence of either α7 or β2 subunits in layers II/III increases pyramidal neuron activity; this is most likely because of a decrease in the cholinergic drive of interneurons directly inhibiting pyramidal neurons.

Because α5 subunits are expressed in cortical layers that were outside of our imaging field in layers II/III [START_REF] Poorthuis | Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors[END_REF] , we selectively re-expressed the WT α5 subunit in the PFC layer II/III interneurons of α5KO-GADCre mice by using a conditional lentiviral vector (Fig. 2a,b). We observed full restoration of pyramidal neuron activity to control levels after re-expression of the WT α5 subunit, whereas, when the α5 subunit encoding the human SNP was re-expressed in α5KO-GADCre mice, pyramidal cell activity was only partially restored, to levels between those of control mice and α5KO-GADCre mice (Fig. 2c). Since PV interneurons do not express the requisite β2 subunit to form a functional α5-containing nAChR, these results may be due to the expression of the α5 subunit in either VIP or SOM interneurons. Because the activity in pyramidal neurons is restored, re-expression of α5 in VIP interneurons likely represents the dominant effect on the circuit behavior. These data indicate that the hypofrontality in α5KO and α5SNP mice can be attributed exclusively to an effect on layer II/III GABAergic interneurons.

In addition, because β2 subunits are also expressed in other cortical layers [START_REF] Poorthuis | Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors[END_REF] , we monitored pyramidal cell activity in β2KO mice after re-expression of the β2 subunit. WT β2 subunit was re-expressed along with tdTomato as a reporter by targeted injection of a β2-IRES2-tdTomato bicistronic lentiviral vector into layers II/III of the PrLC of β2KO mice (Fig. 2d and Supplementary Fig. 7). In the PrLC, only the interneurons in layers II/III express nAChR α subunits, and therefore the lentivirus-mediated expression resulted in functional nAChRs containing β2 subunits exclusively in interneurons [START_REF] Mcgehee | Physiological diversity of nicotinic acetylcholine receptors expressed by vertebrate neurons[END_REF] . We found that the neuronal activity of β2KO mice was elevated when compared with that in WT mice, and the reexpression of WT β2 led to a normalization of activity (Fig. 2e,f and Supplementary Fig. 8).

These results are consistent with layer II/III re-expression of β2 in SOM and VIP interneurons. When considered together, these manipulations demonstrate the critical role of layer II/III interneurons in controlling pyramidal neuron activity within the same layer.

Our results thus far suggest that the decrease in VIP activity in α5KO and α5SNP mice could result from a dishinhibition of pyramidal cells. To directly examine the influence of nicotinic signaling in each interneuron type, we examined the spiking activity of VIP, PV and SOM interneurons under conditions of α5 knockdown. We injected VIP-Cre, PV-Cre, or SOM-Cre with a Cre-dependent AAV vector to direct expression of GCaMP6f to each of these layer II/III interneuron populations. To specifically reduce expression of the α5 subunit in vivo, we used CRISPR-associated Cas9 endonuclease technology [START_REF] Sander | CRISPR-Cas systems for editing, regulating and targeting genomes[END_REF] . We injected the mice described above with a lentiviral vector expressing Cas9 and guide RNA (gRNA) targeting the Chrna5 gene (LV-CRISPR-α5) in layers II/III of the PFC (Fig. 3a,b). Choice of the gRNA sequence and analysis of potential off-target activity by next-generation sequencing (NGS) are described in the Supplementary Methods (Off target analysis using next generation sequencing). The rate of insertions/deletions was quantified to be 15.5% (Supplementary Fig. 9 and Supplementary Tables 2 and3). Using the lentiviral vector in vivo, we found a strong reduction in the median spike frequency of VIP interneurons in VIP-Cre mice injected with LV-CRISPR-α5 lentivirus (3.49 ± 0.66 spikes/min) when compared to control mice injected with a control non-targeting CRISPR lentiviral vector (21.5 ± 2.17 spikes/min; Fig. 3c-f and Supplementary Fig. 10). We then examined how reduction of the α5 subunit in VIP interneurons could affect the spontaneous activity of PV and SOM interneurons, using PV-Cre or SOM-Cre mice together with AAV vectors for Cre-dependent expression of GCaMP6f. We did not observe any significant difference in PV-Cre mice injected with the LV-CRISPR-α5 lentivirus (6.07 ± 0.60 spikes/min) when compared to control mice (5.62 ± 0.45 spikes/min; Fig. 3g-j), which is consistent with weak innervation of PV interneurons by VIP interneurons [START_REF] Pi | Cortical interneurons that specialize in disinhibitory control[END_REF][START_REF] Pfeffer | Inhibition of inhibition in visual cortex: the logic of connections between molecularly distinct interneurons[END_REF] . In contrast, targeted injection of α5-targeting lentivirus into layers II/III resulted in an increase in SOM interneuron spontaneous activity (21.63 ± 0.67 spikes/min) when compared to control mice (5.57 ± 1.31 spikes/min; Fig. 3k-n), whereas pyramidal cell activity was reduced (Fig. 3o-r). Our data show that there is a central role for the α5 subunit in controlling VIP-mediated disinhibition via SOM interneuron inhibition of pyramidal neurons in layers II/III (Fig. 3s).

On the basis of evidence that the nicotine levels obtained from smoking can improve cognitive abilities [START_REF] Levin | Nicotinic effects on cognitive function: behavioral characterization, pharmacological specification, and anatomic localization[END_REF] and can mitigate certain schizophrenic symptoms [START_REF] De Leon | A meta-analysis of worldwide studies demonstrates an association between schizophrenia and tobacco smoking behaviors[END_REF][START_REF] Hambsch | Chronic nicotine improves short-term memory selectively in a G72 mouse model of schizophrenia[END_REF] , we administered nicotine to WT and α5SNP mice by mini-pump infusion and examined alterations in layer II/III neuronal firing before and after infusion (Fig. 4a). The dose of nicotine was selected to maintain a plasma concentration analogous to that observed in smokers [START_REF] Matta | Guidelines on nicotine dose selection for in vivo research[END_REF] . Pyramidal neuron activity significantly increased after 2 d of nicotine administration in WT mice and remained elevated after 2 weeks (Fig. 4b-e and Supplementary Fig. 11). Also, we found that nicotine treatment 1-2 weeks in duration increased pyramidal neuron activity in α5SNP mice to the levels of WT baseline activity (Fig. 4b-e). An explanation for these data is that chronic nicotine exposure desensitizes the β2-containing nAChRs expressed by interneurons. β2dependent, but not α7-dependent, nAChR currents in the interneurons of PFC layers II/III were completely desensitized after exposure to the concentrations of nicotine obtained from smoking in slice preparations [START_REF] Poorthuis | Layer-specific modulation of the prefrontal cortex by nicotinic acetylcholine receptors[END_REF] . To test this in vivo, we administered the same dose of nicotine in SOM-Cre mice injected with AAV encoding Cre-dependent GCaMP6f expression and observed a significant reduction in SOM interneuron activity when compared to that in control mice (Fig. 4f-h). These results suggest that the normalization of neuronal firing in α5deficient mice by nicotine administration is mediated by dominant nicotine-evoked desensitization of the β2-containing receptors on SOM interneurons. As a result, SOM inhibition of pyramidal cells is reduced. Together these data support the conclusion that layer II/III PFC activity is controlled by different nAChR subunits expressed specifically on different classes of interneurons. These experimental results argue strongly that nicotine, or nicotinic agonists, could normalize PFC activity and thus reduce some of the cognitive deficits found in humans with neuropsychiatric disorders.

In summary, we show that endogenous ACh distinctly recruits specific interneuron types within an interneuron hierarchy that controls pyramidal neurons in layers II/III of the PrLC, through differential expression of nAChR subunits. We also provide the first experimental evidence, to our knowledge, that the α5 human polymorphism, which predisposes to nicotine addiction, provokes a decrease in neuronal activity in rodents that mirrors the hypofrontality observed in patients with psychiatric disorders, including schizophrenia and addiction [START_REF] Barch | Selective deficits in prefrontal cortex function in medication-naive patients with schizophrenia[END_REF] .

Further, we demonstrate that chronic nicotine treatment of mice carrying the α5 SNP restores their neuronal activity deficits, and these findings together provide the basis for examining new pharmacotherapeutic strategies. Positive allosteric modulators of defined nAChR subunits, like the polymorphic α5 subunit, are strong candidates [START_REF] Morel | Nicotine consumption is regulated by a human polymorphism in dopamine neurons[END_REF] . Finally, our results support the hypothesis that individuals with polymorphic CHRNA5 smoke to self-medicate in order to ameliorate at least some symptoms of cortical dysfunction.

METHODS

Subjects. Male α7 -/-(α7KO) [START_REF] Orr-Urtreger | Mice deficient in the alpha7 neuronal nicotinic acetylcholine receptor lack alpha-bungarotoxin binding sites and hippocampal fast nicotinic currents[END_REF] , β2 -/-(β2KO) [START_REF] Picciotto | Abnormal avoidance learning in mice lacking functional highaffinity nicotine receptor in the brain[END_REF] , α5 -/-(α5KO) [START_REF] Salas | The nicotinic acetylcholine receptor subunit alpha 5 mediates short-term effects of nicotine in vivo[END_REF] and WT C57BL/6J mice used in this study were maintained at Charles River Laboratories, and experiments were performed Mice engineered to harbor the α5 D398N variant (α5SNP mice) were obtained via homologous recombination. They were generated by the Ingenious Targeting Laboratory using C57BL/6 embryonic stem cells and genomic constructs derived from a C57BL/6 BAC library. The targeting construct was mutated to convert an aspartic acid codon (GAT) to an asparagine codon (AAT) at amino acid position 397 of the mouse α5 subunit gene (Chrna5).

The amino acid position 397 in mice is equivalent to position 398 in human CHRNA5. The neomycin selection cassette of the targeting construct was removed by breeding α5SNP mice to a C57BL/6J strain expressing a Zp3-cre transgene (C57BL/6-TgN(ZP3-Cre)93Knw, Jackson Laboratories). Removal of the neomycin cassette was verified by PCR. GAD67-Cre mice were published previously [START_REF] Fuchs | Genetically altered AMPA-type glutamate receptor kinetics in interneurons disrupt long-range synchrony of gamma oscillation[END_REF][START_REF] Tolu | A versatile system for the neuronal subtype specific expression of lentiviral vectors[END_REF] . To generate GAD67-Cre/tdTomato mice, GAD67-Cre mice were crossed with a ChAT-Cre/tdTomato line [START_REF] Arroyo | Prolonged disynaptic inhibition in the cortex mediated by slow, non-α7 nicotinic excitation of a specific subset of cortical interneurons[END_REF] . GAD67-Cre mice were bred onto a homozygous α5KO background to obtain α5KO-GADCre mice.

To generate VIP-Cre/tdTomato mice, homozygous VIP-IRES-Cre (VIPtm1(cre)Zjh, Jackson Laboratory) mice were crossed with homozygous Ai9 mice (B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J, Jackson Laboratory) as previously described [START_REF] Prönneke | Characterizing VIP Neurons in the Barrel Cortex of VIPcre/tdTomato Mice Reveals Layer-Specific Differences[END_REF] .

PV-Cre (008069, Jackson Laboratory) and SOM-Cre (013044, Jackson Laboratory) mice were bred onto a C57BL/6J background.

All experiments were performed using male mice.

Social interaction tests.

Eight-to 12-week-old male WT and α5SNP littermates were tested for social behavior using a three-chamber social approach paradigm [START_REF] Choi | The maternal interleukin-17a pathway in mice promotes autism-like phenotypes in offspring[END_REF] . Experimental mice were habituated for 1 h in separate, clean holding cages and then were introduced into a threechamber arena with only empty object-containment cages (circular metallic cages, Stoelting

Neuroscience) for a 10-min acclimation phase in two 5-min sessions in a 3-to 4-h period (habituation periods I and II). The following day, the mice were placed in the center chamber (without access to the left and right social test areas) and were allowed to explore the center area for 5 min (open field period). After this exploration period, the barriers to the adjacent chambers were removed, allowing mice to explore the left and right arenas, which contained a social partner (i.e., unfamiliar C57BL/6J male mouse) in one chamber and an inanimate object (plastic toy) in the other chamber. Experimental mice were given 10 min to explore both chambers and were measured for approach behavior as interaction time (i.e., sniffing, approach) with targets in each chamber (within 2 cm, excluding non-nose contact or exploration). Sessions were video recorded, and object exploration time and total distance moved were analyzed using the Noldus tracking system. A social preference index was calculated as the percentage of time spent investigating the social target (unfamiliar C57BL/6J male mouse) out of the total exploration time for both objects. The analysis was conducted with investigators blinded to the genotypes of subjects. Arenas and their contents were thoroughly cleaned between testing sessions. Multiple mice from different home cages were used as social targets to prevent potential odorant confounds from target home cages. Stereotaxic injections and chronic cranial windows. Mice were anesthetized with ketamine (10% in PBS; Imalgen 1000, Rhone Mérieux) and xylazine (2% in PBS; Rompun, Bayer), 10 ml/kg intraperitoneally (i.p.), and placed into a stereotaxic frame. Mice were kept on a thermal blanket, and their eyes were protected with artificial tear ointment. The scalp was washed with three alternating swabs of 70% ethanol and betadine. Surgical tools were sterilized using a hot glass-bead sterilizer. The skull was exposed, and 1% xylocaine was applied to the periosteum. A chronic cranial window was prepared as previously described [START_REF] Holtmaat | Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window[END_REF] (Supplementary Fig. 1). Briefly, the skull was gently thinned using a dental drill around the region of interest, and the thinned bone was removed using forceps, leaving the dura intact.

Prepulse inhibition of

For recordings of pyramidal neurons, 200 nl of AAV virus (2.2 x 10 13 genome copies (GC)/ml; AAV1.syn.GCaMP6f.WPRE.SV40, University of Pennsylvania Vector Core) was injected bilaterally at the following coordinates-PrL: AP, +2.8 mm from the bregma; L, ±0.5 mm; DV, -0.3 to -0.1 mm from the skull-using a Nanoject II (Drummond Scientific) at the slow infusion setting. For recordings of interneurons in the Cre lines, 200 nl of AAV virus (2.1 x 10 13 GC/ml; AAV1.Syn.Flex.GCaMP6f.WPRE.SV40, University of Pennsylvania Vector Core) was injected bilaterally at the same coordinates as described above. The glass pipette was left in situ for an additional 5 min, before being slowly withdrawn from the brain.

The optical window was covered with a circular coverglass (5 mm in diameter) and was sealed to the skull with dental cement (Coffret SUPERBOND complet, Phymep). A sterile small stainless steel bar with a screw hole was embedded into the acrylic over the cerebellum to head-fix the mouse for imaging sessions, and all exposed skull was covered with dental cement.

Lentiviral expression vectors and stereotaxic procedure. The lentiviral expression vectors

were derived from the pHR' expression vectors first described by Naldini et al. [START_REF] Naldini | In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector[END_REF] with several subsequent modifications. In the lentivirus used in this study, expression of mCherry cDNA is under the control of the mouse phosphoglycerate kinase (PGK) promoter. Further details can be found in ref. 45. Briefly, to create the pLV-PGK-mCherry-WPRE vector, a two-step strategy was used. First, mCherry cDNA was amplified from the pRSET plasmid (a kind gift of the laboratory of S. Tajbakhsh, Stem Cells and Development lab, Institut Pasteur, Paris, France), using the following primers-forward, 5′-GTACTCGAGCCACCATGGTGAGCAAGGGC-3′; reverse, 5′-GCTGACGCGGCCGCTTACTTGTACAGCTCGTCCATG-3′-and the T/A cloning strategy was used to introduce mCherry cDNA into the pGEM-T Easy vector (Promega).

Next, we took advantage of the XhoI site in the multiple-cloning region of the vector to replace the fragment encoding eGFP in the preexisting pTRIP-PGK-eGFP lentiviral vector [START_REF] Maskos | Nicotine reinforcement and cognition restored by targeted expression of nicotinic receptors[END_REF] with mCherry cDNA between the XhoI and BsrGI sites. This viral vector was used as a control injection for the β2 rescue experiments.

To create the pLV-dsmPGK-β2-IRES-tdTomato-WPRE vector, we used a previously described PTP-β2-IRES-GFP-WPRE vector [START_REF] Maskos | Nicotine reinforcement and cognition restored by targeted expression of nicotinic receptors[END_REF] and replaced the full IRES-GFP sequence with IRES-tdTomato available from Clotech (pLVX-IRES-tdTomato) by using the Gateway system. Lentivectors were diluted in PBS before stereotaxic injection to achieve a dose corresponding to 250 ng of p24 protein in 2 µl.

Production of the conditional lentivectors was performed as previously described [START_REF] Morel | Nicotine consumption is regulated by a human polymorphism in dopamine neurons[END_REF] . Briefly, viral particles were generated by cotransfection of HEK-293T cells with the vector plasmid, a packaging plasmid and an envelope plasmid using the calcium phosphate protocol. At 2 d after transfection, viral particles were collected in the supernatant, treated with DNase I and MgCl 2 , passed through a 0.45-µm filter, concentrated by ultracentrifugation and resuspended in a small volume of PBS. Viral stocks were stored in small aliquots at -80 °C before use.

Viral titers were estimated by quantification of the p24 capsid protein using the HIV-1 p24 antigen immunoassay (ZeptoMetrix) according to the manufacturer's instructions.

Lentivectors were diluted in PBS before stereotaxic injection to achieve a dose corresponding to 150 ng of p24 protein in 2 µl for re-expression of the α5 subunit. As controls, we used vectors carrying only eGFP under the same promoter and we injected the same volumes. The lentiviral vectors were injected bilaterally into the PrLC at the following coordinates-PrL: AP, +2.8 mm from the bregma; L, ±0.5 mm; DV, -0.25 to -0.1 mm from the skull-using a Nanoject II (Drummond Scientific) at the slow infusion setting.

CRISPR/Cas9 nuclease RNA-guided genome editing. We used CRISPR-associated Cas9 endonuclease technology for inactivation of the α5 subunit (Sigma-Aldrich). Three different gRNAs were designed by Sigma-Aldrich using an algorithm based on Bowtie sequence analysis software. gRNAs targeting a site in the first two-thirds of the coding sequence with at least 3 bp of mismatch with any other location in the host genome were proposed for testing by genomic cleavage detection assay (CEL-1 endonuclease). Assessment of the CRISPR/Cas9 on-target and off-target effects is described in the Supplementary Methods (Off target analysis using next generation sequencing) and Supplementary Tables 2 and3.

After screening of three target sites in the Chrna5 gene, the following target site was chosen:

TGTCACTCAGGTGTTCCACAGG. The lentiviral vector uses a one-plasmid system consisting of an EF1-α-driven Cas9 gene and a U6-driven gRNA, with both puromycin and GFP coexpressed with Cas9. 2.5 µg of U6-gRNA was nucleofected with 5 µg of Cas9 plasmid. In vitro validation was performed in mouse N2a cells (Sigma-Aldrich), where the targeting efficiency was established to be 13.2% by enzymatic assay. N2a cells tested negative for mycoplasma. Insertions and deletions were detected at the gRNA target via mismatch assay (CEL-1 endonuclease) using the following primers: forward, 5′-GATTATTGATAACATTGAGTTTC-3′; reverse, 5′-CTCTAGTGGCTAACTCACAC-3′. As a control, we used a lentiviral plasmid vector that includes a gRNA sequence that does not target known human, mouse or rat genes (CRISPR12-1EA, Sigma-Aldrich). Viral particles were generated by cotransfection of HEK-293T cells with the vector plasmid, as described above. For the α5-Cas9 lentiviral vector, 49 ng of p24 protein in 1 µl was locally injected in layers II/III of the PFC, whereas the control virus was diluted in PBS before stereotaxic injection to achieve a dose corresponding to 50 ng of p24 protein in 1 µl. In addition, 100 nl of AAV1.CAG.tdTomato.WPRE.SV40 (1.52 x 10 13 GC/ml; University of Pennsylvania Vector Core) was added to the solution to allow localization of the cells transduced with LV-CRISPR virus.

Mouse handling for awake imaging. Mice were habituated to the imaging environment by handling and training for 4 d, as described previously [START_REF] Lepousez | Odor discrimination requires proper olfactory fast oscillations in awake mice[END_REF][START_REF] Guo | Procedures for behavioral experiments in head-fixed mice[END_REF] . Briefly, the animals were habituated to the following conditions sequentially: (1) gentle handling, (2) free exploration of a 50-ml open-ended support tube (for the mouse to rest in during imaging), (3) incremental periods of mild head fixation in which the mouse's head was briefly secured to the head post (5 s to 15 s) and ( 4) extended periods of head fixation on the mouse stage while the mouse rested in the support tube (5 min to 60 min).

Immunofluorescence. At the end of the final two-photon imaging session, immunofluorescence was performed in order to verify the injection site. Fixation was carried out by transcardiac perfusion of anesthetized mice using 4% paraformaldehyde (PFA), and the brains were removed and post-fixed by immersion in PFA for 2 d at 4 °C. The brains were then immersed in 30% sucrose in PBS overnight for cryoprotection at 4 °C. Serial 40-µm coronal sections were cut with a sliding microtome (Leica Microsystems), transferred to PBS, mounted on slides and coverslipped with ProLong Gold antifade reagent with DAPI (Invitrogen). Microscopy was carried out with a Zeiss epifluorescent microscope.

To determine expression patterns in β2KO-GADCre mice injected with β2-IRES2-tdTomato and in α5KO-GADCre mice injected with PDGF-floxed-mCherry-α5 WT/SNP-IRES2-eGFPP-WPRE, we anesthetized and perfused the mice as described above. Serial 40-µm coronal sections were cut with a sliding microtome, collected and washed in PBS treated with 0.2% Tween-20 (PBS-T), immersed in 10% normal horse serum blocking solution, and incubated overnight with primary antibodies against GFP (1:2,000; Abcam, ab38689) and RFP (1:2,000; Molecular Probes, R10367) followed by incubation with secondary antibodies conjugated to Alexa Fluor 488 (green; 1:500; Jackson ImmunoResearch Laboratories, 715-545-150) and Alexa Fluor 568 (red; 1:500; Abcam, ab175470). The sections were mounted on slides and coverslipped with ProLong Gold antifade reagent with DAPI. Microscopy was carried out with a Zeiss LSM-700 confocal microscope (Supplementary Fig. 7). For all antibodies, validation is provided at the manufacturer's website.

Surgical implantation of mini-pumps. Mice were anesthetized with a xylazine and ketamine combination (15% xylazine/2.5% ketamine/82.5% PBS). Osmotic mini-pumps (model 2004, ALZET DURECT) were implanted subcutaneously at the nape of the neck as previously described [START_REF] Besson | (j) Median firing rates for PV and PV + LV-CRISPR-α5 mice. No significant difference was found between the groups (P = 0.55, Kruskal-Wallis test) n = 4 mice for each group. (k) Representative in vivo two-photon image of GCaMP6f-expressing SOM interneurons in layers II/III of the PFC of SOM-Cre mice. Scale bar, 50 µm. (l) Representative spontaneous Ca 2+ transients of SOM interneurons and SOM + LV-CRISPR-α5 interneurons and their corresponding raster plots[END_REF] . Nicotine or saline was delivered at a constant rate (2.4 mg per kg body weight per day for nicotine).

In vivo two-photon imaging. In vivo imaging was performed with an Ultima IV two-photon laser-scanning microscope system (Bruker), using a 16x, 0.8 N.A. water immersion objective (Nikon) with the femtosecond laser (MaiTai DeepSee, Spectra Physics) tuned to 950 nm for imaging of cells expressing GCaMP6f and to 1020 nm for imaging of cells expressing tdTomato. Time-series movies of neuronal populations expressing GCaMP6f were acquired at the frame rate of 6.926 Hz (182 x 182 µm field of view; 0.71 µm/pixel). The duration of each focal plane movie was 216.564 s (1,500 frames) to track spontaneous neuronal activity. For interneuron recordings, time-series movies of interneuronal populations expressing GCaMP6f

were acquired at the frame rate of 30.33 Hz (405 x 405 µm field of view; 0.79 µm/pixel). The duration of each focal plane movie was 164.897 s (5,000 frames).

Randomization and blinding.

The mice from all experimental groups, both baseline and nicotine treated, were imaged in a random order. Experiments were performed with blinding for the β2 re-expressing mice and for recordings of the VIP-Cre and PV-Cre baseline and SOM-Cre nicotine-treated mice but not for the rest of the experiments. Analysis was performed with blinding for the social interaction test data, the activities of the α5KO, α5SNP and β2 re-expressing mice, and recordings of the baseline PV-Cre and SOM-Cre mice and SOM-Cre nicotine-treated mice, but not for the rest of the experiments.

Two-photon data analysis. Image analysis was performed offline with

  at 3 months of age. The experiments described in the present work were conducted in accordance with the guidelines on the ethical use of animals from the European Community Council Directive of 24 November 1986 (86/609/EEC) and in accordance with institutional animal welfare guidelines and were approved by Animalerie Centrale, Médecine du Travail and the Ethics Committee CETEA of Institut Pasteur, protocol numbers 2013-0056, 2013-0104 and 2015-0007.

  the startle reflex. Male WT and α5SNP littermates ranging in age from 60-75 d were used for the study. The PPI apparatus (SR-Lab, San Diego Instruments) consists of a sound-attenuating chamber and a vented Plexiglas animal enclosure (4 cm in diameter) that rests on a 20 x 13 cm Plexiglas frame. Background (70 dB), prepulse (74, 78, software (San Diego Instruments) and presented through a speaker mounted (13 cm) above the animal enclosure. The magnitude of the startle response was measured by a piezoelectric accelerometer mounted below the Plexiglas animal enclosure.Each session was initiated with a 5-min acclimation period followed by initiation of the test session with five pulse-only trials (white noise, 120 dB/40 ms); each session also concluded with five identical pulse-only trials. Both the pre-and post-pulse trials were excluded from analysis. The remaining trials consisted of pseudorandomized presentation within ten trial blocks of a blank (no-stimulus) trial, a pulse-only trial and four different types of prepulse + pulse trials (20 ms at 74, 78, 82 or 90 dB followed by a pulse (120 dB) 100 ms after the prepulse). PPI was calculated using the means of ten trials for each type of stimulus: percent PPI = (pulse-only score -(prepulse + pulse score))/pulse-only score x 100.

Figure 4 Figure 1 Figure 2 Figure 3 Figure 4

 41234 Figure 4 Effect of chronic nicotine on the prelimbic cortex activity of WT and α5SNP mice

  Two-photon data. Kruskal-Wallis one-way analysis of variance combined with multiple-comparison testing was applied on the aggregated activity (transients/min and spikes/min) of the neurons in all mouse groups in order to study the statistical similarities between them. For each mouse category, we used interpolation in order to account for the different number of neurons recorded in the different animals. In this way, our statistical analysis assigned the same weight to each neuronal population recorded in the different mice of a given category. Levene's test was used to compare the variances of the neural distributions that were similar between groups that were statistically compared. We used Welch's F-test ANOVA as a complementary test for heteroscedasticity. This test gave similar results as the Kruskal-Wallis test with the same level of significance. We used the multiple-comparisons Tukey-Kramer test in order to determine which pairs of mean ranks were significantly different. Animals were not excluded from the analysis. The number of mice needed for our study was determined by preliminary experiments in anesthetized mice and pilot experiments in awake mice. Box plots, histograms and empirical cumulative distribution functions (Kaplan-Meier estimate) were used to compare the different distributions of neural activity between the mouse types. For each box, the central mark is the median, the edges of the box are the 25th and 75th percentiles, and the whiskers extend to the most extreme data points not considered outliers. The ends of the whiskers represent the lowest data value still within 1.5 times the IQR (interquartile range) of the lower quartile and the highest data value still within 1.5 times the IQR of the upper quartile. On the empirical Behavioral data. Statistical analysis was performed using Prism (GraphPad) software. Social interaction task differences were assessed using unpaired Student's t test. Because the variance between genotypes for the social index and interaction time measures were not equal, unpaired Student's t test with Welch's correction was used. Two-way repeatedmeasures ANOVA with Sidak post hoc test was used for the PPI data analysis. All data other than the aforementioned social interaction tests met the assumptions of the chosen statistical test: normality and equal variances. neuronal activity due to decreased VIP interneuron inhibition of SOM interneurons in layers II/III. Adapted from ref.25 with permission of Nature Publishing Group. All control mice were injected with a CRISPR nontargeting control lentiviral vector (Online Methods).

	ImageJ software. The Image Stabilizer Statistical analysis. cumulative distribution plots, 95% lower and upper confidence bounds were computed with time series were corrected using
	(http://www.cs.cmu.edu/~kangli/code/Image_Stabilizer.html). Regions of interest (ROIs) Greenwood's formula.
	were manually selected in FIJI, and detection of Ca 2+ transients of individual neurons was
	performed automatically by using a custom-written toolbox in MATLAB (Mathworks). For
	details on the analysis method, see the Supplementary Methods (Two-photon data analysis).

Acknowledgments F.K. is a scholar of the Pasteur Paris University Doctoral Program (PPU) and received a stipend from the Stavros Niarchos Foundation. This work was supported by the CNRS UMR 3571, the Fondation de la recherche médicale (FRM grant DPA20140629803), the Agence nationale de la recherche (ANR), the Laboratoire d'Excellence BIO-PSY (including salary support to F.K., AAP Fin de thèse 2015), the programme PasteurInnov 2012, by the FP7 ERANET programme NICO-GENE, Grant Agreement n009 BLANC 20092009BLANC 20, by the European Commission FP7 RTD

Data availability. The custom-written toolbox in MATLAB (Mathworks, 2014b) is available upon request. All data and materials used in this study are available upon request.


Silvia Serrano

FROM CULTURE TO CULT: MUSEUM COLLECTIONS AND RELIGION IN CONTEMPORARY GEORGIAN NATIONAL DISCOURSE

In 1999, the Georgian Ministry of Culture, the Baltimore Walters Art Gallery and the Foundation for International Arts and Education decided to exhibit 160 artefacts, including sacred objects, from Georgian collections in American museums. A small circle of students in Tbilisi, the capital of Georgia, started a protest action against the exhibiting of sacred artefacts, soon gaining support from the political opposition leaders and a large part of population. As a result, the authorities decided to cancel the exhibition. Based on the case study of these protest actions, this chapter aims to show the ways in which modern-day Georgian nationalism has been remodeled around religious categories, and the political and social implications of this remodeling.

The chapter argues, that the impoverishment of the specific religious meaning of Christianity during the socialist rule has provided a background for filling Orthodox Christianity with new meanings and interpretations. In the context of new political developments, this process facilitated the re-emergence of Christianity as the central aspect of the national and political discourse. In present-day Georgia religion appears to engage discourses and issues which go beyond religious matters, such as those of foreign policy, political orientation, corruption, and national identity. Such re-interpretation of Christianity in the contemporary context has contributed to the process of de-secularization of the society.

In 1999, the Georgian Ministry of Culture, the Baltimore Walters Art Gallery and the Foundation for International Arts and Education planned an exhibition called "Land of Myth and Fire: Art of Ancient and Medieval Georgia." One hundred and sixty items from the several Georgian museums such as the Simon Janashia Museum, the State Museum of Georgia (The Museum of History) and the Institute of Manuscripts were selected for the exhibition in Baltimore, in San Diego, and Houston. But as soon as this project was made public, a group of students initiated a protest action against the exhibition, arguing that sending the sacred objects abroad would endanger Georgia by depriving the country from the protection they provide.1 Within a few months, the protest had grown and gained support from the political opposition leaders and a large part of the population. As a result, eventually the authorities decided to cancel the exhibition (Dobrzynski, 1999).

Using the case study of these protest actions as an example and a historical context, this chapter aims to analyze and show the ways in which the modern-day Georgian nationalism has been remodeled around religious categories and the political and social implications of this remodeling. Indeed, religion has always played a central role in the formation of Georgian national identity, as illustrated by the motto of the well-known writer-politician of the second half of the 19th century, Ilia Chavchavadze, " Language, Fatherland, Faith" [ena, mamuli, sarts'munoeba]. Or, by the struggle of the 19th century intellectuals such as Ilia Chavchavadze, Niko Marr and Iakob Gogebashvili to restore the autocephaly of Georgian Church which was abolished by the Russians (Vardosanidze, 2001; Werth, 2006). Chavchavadze's motto implied the secularized vision of religion, in which religion served as part of cultural identity, tradition and a collective memory. Nevertheless, faith was not the main criteria defining "Georgianness:" Chavchavadze defended Muslim Georgians of Ach'ara, claiming that "faith does not determine nationality-a Georgian always remains a Georgian regardless of his religious orientation" (Chavchavadze, 1965:15). In a similar spirit, Iakob Gogebashvili, the author of the first Georgian handbook for schools, Deda Ena [Mother Tongue], dedicated it to "all Georgians, regardless of faith." But after the declaration of independence on 9th April 1991, new conceptions of nationhood emerged, calling into question the secularized vision of the Nation inherited from the 19th century: religion is no longer interpreted only as "for memory" (Hervieu-Léger, 1993) and tradition. In fact, the Church now plays a central role in the political and public life of Georgia.

Rather than taking for granted a predefined essence of "religion," this chapter will explore rationales of different political and social actors for manipulating with religious symbolism. Hence there are several sub-goals pursued in the different sections of this chapter. The first is to identify the entrepreneurs who promote different models of the nation and the context in which they use the resources provided by the religion. The second sub-goal is to analyze arguments put forward by the Church in its bid to reappropriate the museum collections which were nationalized during the Soviet regime. In the third section I will analyze different meanings attached to artifacts by social actors. The final section will question new and old meanings of these artifacts in order to understand how the Georgian national narrative is being reshaped along the religious line.

ACTORS, STRATEGY AND CONTEXT

The protest began with small student gatherings on April 24. Later during the spring, it spread geographically, involving various social groups (Akhali Taoba, December 1 1999) and becoming more politicized as the demonstrators received support from several opposition leaders and other politicians. In an attempt to calm the protest, Gary Vikan, the director of the Baltimore Walters Art Gallery, and Gregory Guroff, the chairman of the Foundation for International Arts and Education flew to Tbilisi to meet the officials. Commissions were set up to negotiate the matter and reach a compromise. On July 30 1999, the Georgian ambassador to the United States returned to Georgia to participate in the negotiations (Cash, 1999). However, nothing helped; the Catholicos Patriarch made a public announcement about the cancellation of the exhibition in a sermon at the end of July (Sakartvelos Resp'ublik'a, August 27 1999). In the next section I will try to identify the different actors and explore the context for power relationships between them.

From the April to July of 1999, a group of students played a crucial role, organizing gatherings, demonstrations and hunger strikes. Their protest was typical of poorly institutionalized social movements characterized by irregular membership, unstable organizational structure and lack of rational strategy. The students were suspicious of all "institutional" actors, politicians and even the Church. They were constantly concerned about "being manipulated," thus avoiding involvement in politics and contact with political organizations. After the mother of one of the students was beaten by policemen, they expressed their bitterness: Many politicians came to us: Manana Archvadze, Irakli Tsereteli, Valery Kvaratskhelia, Vakhtang Goguadze, Boris Kakubava, Vakhtang Rcheulishvili, Nodar Natadze... She [the mother] ended up with brain concussion and bruises

[…] and the only reason was that she loves her country (Akhali Taoba, 4 June 1999). 2 Nevertheless, the protest grew, and all over the country petitions were collected in support of the students, demanding that the exhibition be cancelled and the treasures transferred from State museums to the Church (Akhali Taoba, June 1 1999). The issue of exhibition soon went beyond the concern of the narrow circles of extremist orthodox believers or staunch nationalist activists and became involved in the broader public debate for the next several months. It was covered by daily newspapers. Years later, the issue was still remembered by students when they were asked about the consequences of sending Georgian museum collections abroad. 3 How did it happen that a few dozen youths, having no social capital and a poor connection with networks of activist political and NGO organizations succeeded in maintaining pressure on the authorities to the extent that the latter finally cancelled the exhibition? The students were seen not as active social actors but as a handful of agitators manipulated by unscrupulous priests and politicians. Indeed, former Zviadists, 4 the veterans of the national movement of the 1980-1990s as well as recent converts to nationalism used the student protests as a political opportunity for promoting their goals.

The first to seize the opportunity was Guram Sharadze. An elected MP in 1995 as a leader of the civil movement "Language, Fatherland, Faith," he chaired the Parliamentary Migrations Committee in 1999, and was known for his nationalist campaigns and campaigns against religious minority groups. 5 He systematically depicted himself as a defender of the nation which, according to him, was in danger. His protest against the exhibition was only one piece of a larger project; he used any controversial political occasion to heat a debate on populist issues. At the same period, he campaigned in favor of stating a person's ethnicity in official documents. He also targeted Christian "sects," advocating the prohibition of Jehovah's Witnesses, encouraging persecutions against them and the burning of their literature (Akhali Taoba, June 9 1999). As soon as the project of exhibition was made public, on April 24, Sharadze organized the first student rally in front of the Tbilisi State University, before moving to S. Janashia Museum and starting a hunger strike (Rezonansi, April 30 1999). The government officials accused him of being the main person behind the student unrest; in proof of this, E. Tevdoradze, an MP from the president Shevardnadze's bloc, commented that she saw some of the students going out from Sharadze's office on the eve of the demonstration.

Other actors, who associated themselves with the earlier Zviadist nationalist movement, used similar symbolic and rhetorical resources in their protest against the exhibition. Among the actors were Nodar Natadze, Manana Archvadze, widow of the former evicted president Gamsakhurdia, and Zviad Dzidziguri, who published an open letter in support of the protesting students from the penal colony, where he was detained at the time. 6 The protestors also included harsh opponents of Zviad Gamsakhurdia, such as the former leader of the extreme liberal wing of the nationalist movement Tamara Ckheidze, and the former speaker of Parliament during 1992-1995, Vakhtang Goguadze. 7 The main goal for these political actors was to reach visibility in the public and political arenas.

Parliamentary elections were to take place in October 1999, followed by the presidential elections in the spring of 2000. G. Sharadze initiated the protest twelve days after founding a new political party, "Georgia above all," created in order to take part and succeed in the elections (Akhali Taoba, April 17 1999). Street pressure was a means to claim his political weight. After July, when he gained a promise from the authorities to include him on the list of the coalition with the Union of the Citizens of Georgia,8 he became much more discreet. He was re-elected in October.9

It should be stressed that the leading political parties (notably the "reformers wing" inside The Union of the Citizens of Georgia) were practically absent from the public debate around the issue of exhibition. Hence the only obvious supporters of the exhibition were the government itself, museum directors and top civil servants. The latter were the main targets of the attacks from the opposition groups. Close to the government, these representatives of the authorities had no personal political ambition; nevertheless their interests were also at stake. It was important for them to protect their positions and advantages which went with them. In defending their position, these public servants used a "scientific" discourse that allowed them to claim their exclusive legitimacy in the questions of national heritage. 10 However, soon they were confronted with another scientific discourse, coming from the lower levels of the professional hierarchy (curators who were in charge of museum collections, restorers, art specialists, etc.). An argument developed among the experts concerning the degree of corrosion of the exhibition items, whether they could endure transportation, opportunities of restoration in the United States etc. 11 Under the technicalscientific cover, this controversy had an indisputably political dimension-groups loyal to Shevardnadze's government versus strongly nationalistically minded groups. 12 The third group involved in the debate consisted of the clergy. The organizer of the exhibition, G. Guroff, underlined the role played by the Catholicos Patriarch, who used all his influence to cancel the exhibition (Dilis Gazeti, May 14 1999). Like the other two political actors, the clergy was not homogeneous. The Church had benefited from the collapse of the Soviet Union: numerous churches had reopened and according to public opinion studies, the Catholicos Patriarch became the most popular figure. Despite this, the Georgian Orthodox Church (GOC) was torn apart by conflict. In 1999, it was a weak institution divided by internal clashes as it had to deal with a low religious and general education level of the most of its priests, corruption, and a rising opposition from radicalized priests and bishops. The Church and the Catholicos Patriarch Ilia II had just avoided a schism by agreeing to leave the World Council of Churches in 1997, thus making compromise with its most radical branch.

The weakness of the Church had two consequences. In the first place, the radical clergymen could act without approval of the Patriarchate. Indeed, while the Patriarchate 10 See, for example, Giorgi Mounjishvili, the head of the Department of the Ministry for Culture, in Dilis Gazeti, May 6 1999; and an interview with Nodar Lomouri carried out by the author in November 2007. 11 See Sp'etsialist'ebi Erovnuli Sagandzuris Shesakheb (1989-1999 ts'lebis masalebi) [Experts on the National Treasury (documents of the 1989-1999)], (1999). Tbilisi. 12 Some experts from the museums joined the demonstrators, sharing with them their concern about the risks of degradation, theft, etc. In the beginning of the 1990s, some pieces, including pictures by Cranach and Chardin, were stolen from the Georgian museums, thus creating another reason for anxiety.

stressed that it was not the initiator of the protest against exhibiting sacred items abroad (Akhali Taoba, May 6 1999), some priests did support the student protests. For example, a priest of the church of Anchiskhat'i joined them on the 4th of May (Rezonansi, May 2 1999). Secondly, the Church did not want a head-on confrontation with the government.

As the demonstrators complained, 13 it was rather passive during the demonstrations, but was eager to seize any opportunity to strengthen its positions.

The main initiators of the protest had a "traditional" secularized vision of the link between the Nation and religion: for them, Orthodoxy was a part of the national culture, and had to be preserved and respected as such. But in using religious references for political goals, some new actors including students and certain members of the clergy, used this opportunity to appear in public arena and to promote an alternative model of religion. Within this model religiosity and Orthodoxy were interpreted not only in terms of the past and culture, but also as central to modern national self-consciousness.

RETURNING PROPERTY TO THE CHURCH

Since Eduard Shevardnadze took power in 1992 after the eviction of the president Zviad Gamsakhurdia, several exhibitions were organized abroad, including sacred art exhibitions, without invoking public protest or any criticism from the Church. 14 Later on, other exhibitions were also organized without any resistance. The 1999 protest actions against the exhibition of religious objects illustrate that the electoral context of the events was crucial in enabling some political actors to play a central role. Even more crucial was the fact that the Church and the State were involved in negotiating relationship with each other.

The forthcoming 2000th anniversary of the birth of the Christ empowered the Church symbolically, 15 whereas an official visit of the Pope, planned in October, gave some room for maneuver in negotiations with the State authorities, for the State needed the Church's approval to the planned visit of the Pope. The political context of 1999 gave the Church several advantages in this process of negotiating power relations with the State. For example, the protest demonstrations provided the Church with a window of opportunity to promote its role as a mediator between the political power and the civil society. In particular, on the 8th of May, Ilia II received a group of demonstrators. On the same day, the day of Saint George, he met the president Shevardnadze. He then declared that he was against the exhibition. However, in order to preserve good relations with the State, he emphasized the President's careful and serious attitude towards the 13 One demonstrator did not understand "why the Patriarch did not ask his congregation to join us" (Akhali Taoba, May 10 1999. issue of sending the treasure. He also agreed, despite the radical clergy's opposition, to invite John-Paul the second to Georgia. 16 One of the arguments put forward to justify the Church's opposition to the exhibition was, as the Patriarchate emphasized, that the "voice of the Church was not taken into account" in its own sphere of competence (Dilis Gazeti, April 26 1999). The Church's claim over the property of the relics and other religious items was indeed a central issue in the negotiations, but it meant a change in the way national treasury was perceived and articulated to the national discourse. The issue of the Churchclaimed property in Georgia is linked to the legacy of the Soviet anti-religious policies as well as of the liquidation of the GOC as a result of the inclusion of Kartli-K'akhetian kingdom into the Tsarist Empire at the beginning of the 19th century. Confessional ties with Orthodox Russia played a crucial role in Georgia's orientation toward Russia which resulted in the signing of the treaty of Georgievsk in 1783, according to which Georgia became a protectorate of Russia. However, Russia later annexed Georgia. It also abolished the autocephaly of the Georgian Church and subjected it to the Russian Church as early as in 1811; the Catholicos-Patriarch of Georgia was replaced by an exarch appointed from Russia, whereas the possessions of the Church were passed under the supervision of the Treasury of the Empire, hence became the property of Russian Empire. 17 Links with Russia, Westernization as well as the development of nationalliberation movement against the Russian colonialism facilitated the development of national consciousness and secularization of the society and religion, which in turn helped to transform religious relics into secular cultural symbols of the nation. The Tsar administration's attack at the Georgian Church thus increased the Georgian nationalists' urge to make the defense of their religious and cultural heritage a top priority.

As elsewhere in the world, the emergence of museum collections paralleled the emergence of modern nationalism. For historical reasons, the "patrimonialization" of ecclesiastical objects played a crucial role in the Georgian national narrative (Gordadze, 2006). In spite of the opposition from Russian exarches, an ecclesiastical museum was created inside the Sioni cathedral in Tbilisi in 1889, in order to save the ancient manuscripts and other religious items (Grdzelidze & Fisher, 2006:159). At the end of 1920, by the time when autocephaly was restored, 18 the relations between the Mensheviks 19 and the Church deteriorated. The clergy began to sell the treasure of the 16 See Rezonansi, May 8 1999, Sakartvelos Resp'ublik'a, May 8 1999. 17 Apart from the first exarch, Varlam Eristavi (1814-1817), all exarches were Russian, and Georgian language was replaced by the Church Slavonic as a liturgical language. During the restoration work between 1830 and 1840, some churches were deteriorated and ancient frescoes covered by lime. 18 Autocephaly was restored in March 1917. 19 The social-democratic party was elected in 1918 and ruled the country until the Sovietization in 1921.

Gelati monastery near the west Georgian city of Kutaisi. As a response to this the government took all the treasures of the Church under its control, in spite of the protests of the Patriarch Leonid (Peters, 1988:290-291). Facing the advance of the Bolsheviks, the social-democratic (Menshevik) government headed by Noe Jordania decided to send the treasures abroad (Vardosanidze, 2001:45). On 23rd of November 1923, during the newly established Soviet rule when the church property was nationalized (Vardosanidze, 2001:59; Musin, 2006:29), the Commissioners of the People of the Republic of Georgia adopted a resolution according to which items of a historical importance belonging to the Church would be transferred to the Museum of Popular Education (Vardosanidze, 2001:75-76).

After the 1943 and WWII a shift occurred in the Soviet religious policy, 20 and the GOC requested that the objects kept in museums be given back to the Church. After Stalin's death, the Catholicos Patriarch Melkisedek (1952-1960) managed to return the remains of St David and St Constantine to the monastery of Motsameta in Kutaisi (Grdzelidze & Fisher, 2006:223). Gorbachev's new political-economic course of Perestroika provided new opportunities for the claims of the Church. The Patriarch canonized some of the persons who helped to save the possessions of the Church, such as Ekvtime Taqaishvili or Father-Superior (Ighumen) Ekvtime Kereselidze, who had hidden the sheet music of liturgical songs and ancient manuscripts from the Soviet regime. 21 Even before the independence was declared formally in 1990, the ecclesiastic property was given back to the Church. According to the resolution of the Cabinet of Georgia of the 12th April 1990, all orthodox churches on the territory of Georgia, the land on which they were built as well as the movable property in the churches belongs to the Patriarchate (Papuashvili, 2003). Nevertheless, the process of returning the property to the Church has been rather obscure and controversial, without precise lists of the possessions and their status, including some objects which it had never owned (Abashidze, 2006). Official texts adopted afterwards left many points unresolved, opening the way for a dispute around the issue of who were the legitimate owners of the museum collections and where they should have been kept. As in other post-Soviet countries, the Church's claims on the cultural heritage have thus become a key issue in the shaping of a new national consensus (Musin, 2006).

The opening of a Church museum became a priority on the Church agenda. As pointed out by the Church's press secretary, Father David Sharashenidze, The conservation of worship items in museums is a short-lived phenomenon. The question of their return to their legitimate owner is soon going to arise. The Church will probably soon have its own museum […]. After the creation of such a museum, the Church will request all the objects which are kept in museums today (Akhali 7 Dghe, April 30-May 6 1999).

The constitutional agreement adopted on 14th October 200222 actually returned the property to the Church, including churches, monasteries, the land as well as all the religious treasuries kept in state museums (The Church and Civil Society in Georgia at the beginning of the XIX century, 2001). The agreement did not specify whether the religious items should be kept in state museums or in the churches, and laws adopted afterwards (law on the import and export of cultural goods adopted on 22nd June 2001, amended in 2003, 2006, and 2007, and the law on Cultural Heritage adopted on 8th May 2007) did not solve all the questions linked with the Church property.

During the protest actions against the exhibition in April 1999, various social groups attempted to give significance to the museum collections on the basis of symbolic as well as social, political and financial benefit that could be gained from them. The issue at stake was to identify who was their legitimate owner (the Georgian Orthodox Church, other Churches, or the State), and whether these artifacts were valuable or sacred objects, whether they were to be regarded as part of a cultural heritage or as liturgical objects. The restitution of the museum collections to the Church meant that they needed to be "denationalized," which was possible only by means of reshaping the national discourse along new lines, which I am going to discuss in the next section.

NATIONAL HERITAGE OR RELIGIOUS HERITAGE?

The interpretation of the museum items within the national and religious discourses illustrates the intertwining between politics, culture and religion. They can be seen as valuable goods for their esthetic qualities, as a testimony of the glorious past, or as religious items. 23 The labeling of the items is a key issue for the actors competing for the status of a legitimate ownership of the Church treasures. Art historians and curators defined these items as "pieces of art" the place of which is a museum, whereas zviadist Z. Dzidziguri argued "that these relics are not museum pieces; for a believer they have much more meaning than a purely esthetic value attached to them by a specialist" (Akhali Taoba, May 10 1999). As the Patriarchate press secretary stressed, "The Church comments that learning about the Georgian culture occurs mainly through religious collections" (Akhali 7 Dghe, April 30-May 6 1999).

The clergy, the students, and even the politicians never mentioned the artistic value of the items. They are seen as "cultural heritage" not because of their esthetic value, but because they are assimilated with the national history. 24 When the students were banned from organizing protest demonstrations in front of the Patriarchate, they withdrew to the monument dedicated to the Georgian language (Deda Ena-Mother Tongue), thus confirming their nationalist orientation. The protestors accused the advocates of the exhibition in disrespecting the value of the nation. For example, an open letter was sent by Bidzina Tvaradze, the president of the "Society for the Protection of Nature and Historic Treasures" to Levan Berdzenishvili, then the director of the National Library who supported the idea of the exhibition, asking "How can a harsh opponent of national consciousness be a director of the National Library" (Lit'erat'uruli Sakartvelo, August 6-13 1999). In this sense, the protest was very similar to the national movement of the 1980s, in which associations for the defense of cultural and religious heritage played an important role (Goujon, 2002). 25 The religious and the national meanings were intertwined in the interpretation of the items to be exhibited. Even though technical arguments such as the risk of corrosion were put forward in order to refute the idea of the exhibition, emphasis was laid on the national as well as religious value of the collection. One student explained that the treasure "is what binds the nation to the Church and God." They blamed the authorities for perceiving the heritage as exclusively national, deprived of its religious significance for the nation. One of the demonstrators complained that "our people have forgotten that those in power nowadays are the communists for whom Georgian Treasury-icons, crosses, etc. are only museum artifacts and not the sacred objects" (Akhali Taoba, May 16 1999). Mamuka Giorgadze, the president of the parliamentary fraction "Sakhalkho" [literally "of the people"] declared that "a government which views an icon as an object can do no good" (Asaval-Dasavali, May 10-16 1999).

According to the press and interviews, this was an exhibition of the sacred Christian art. However, as shown in the album published to accompany the exhibition (Soltes, 1999), most of the items to be exhibited preceded the Christianity and originated in prehistory or in pre-Christian Greek antiquity. These non-Christian items were hardly mentioned in the debate for they would distract from the Orthodox Christian overtones of this debate. The national character of the items was thus largely determined according to their relation to Christianity. Therefore, the objects of cultural heritage such as remains of the saints, icons, worship items, etc. were viewed as national treasure, while other items such as pre-Christian antiquities lost their national relevance because they were not related to Christianity. But at the same time, it was never said that the demand to cancel the exhibition concerned only icons; the Ministry of Culture stressed that the students even rejected a proposition to send any museum items abroad (Akhali 7 Dghe, April 30-May 6 1999), as if the qualities granted to icons were spread to all the items. This particular insistence of the students demonstrates that students were against the exhibition of the museum collections not only because of their Christian contents, but also because of their secular qualities signifying the memory of the nation. Indeed, there is a significant difference between the meaning traditionally given to religious relics or icons and the meaning given by the student-protestors to these museum items. The role they were supposed to play in the physical (standing strong against the enemy) and symbolic (illustrating Georgia's cultural heritage) preservation of the Nation was much more relevant than the power these items had acquired through their religious function and prayers. 26 While the national and the religious visions converged in the recognition of the value of museum collections, the different actors prioritized one of the two meanings and interpreted the relationship between the nation and religion differently from each other. While the politicians (zviadists, G. Sharadze, etc.) considered the heritage as sacred because it was national, for the others (the Church and the students) it was sacred first and foremost because it was linked to the religion and only via this link, to the national identity. However, precise division between the two interpretations was blurred. Moreover, the interpretations were redefined at each protest occasion, the fact which illustrates dynamism and ambiguity of the perceptions of religion and nation in the society.

OLD AND NEW FUNCTIONS OF THE SACRED OBJECTS

To understand why a public debate arose regarding the exhibition, the discussion in this essay needs to be positioned in the wider context of the collapse of the Soviet Union and the following political and social changes. The 1999 events and debate around the issue of exhibition occurred in this transitional period, and were characterized by the current weakness of the state, a shift to capitalist globalization and market economy, and by the reshaping of collective identity. The sacred objects were perceived as an essential element of this collective identity, and were seen as a means for the protection of national consciousness from new threats brought by globalization and the new, capitalist economic and social relations.

There was an acceleration of globalization in 1999 that made it necessary to think about geopolitical identity and orientation of the country. The joining of international community, symbolized by the admission of Georgia to the Council of Europe in April of 1999, was seen both as an opportunity and as a new source of uncertainty. A clear disagreement emerged between those who thought that they could take advantage of globalization and relations with the West and those who saw it as a threat. President Shevardnadze made his first public announcement about the exhibition from Washington, where he was attending a NATO summit. 27 By organizing this prestigious event, its instigators aimed to create a positive image of the country, in a typical example of cultural diplomacy (Sakartvelos Resp'ublik'a, May 1 1999). The "official" intelligentsia understood the benefit of such an international exhibition both for their country and their career development. 28 They feared that the cancellation of the exhibition would contribute to a further international isolation (Dilis Gazeti, May 6 1999)-the first step of which was the GOC's withdrawal from the World Council of Churches. 29 The Patriarchate's stand in this debate was ambiguous. It was interested in promoting an international image of the nation, notably because it would provide it with a legitimacy needed for its relations with the Moscow Patriarchate and the Ecumenical Patriarchate of Constantinople. 30 Therefore, it claimed that, while theoretically it was not opposed to exhibitions abroad, it was against this particular exhibition for specific reasons: the items must stay in Georgia for the 2000th anniversary of Jesus Christ; the number of objects of exposition was too large; there were technical problems related to the maintenance of the items, etc. (K'viris P'alit'ra, August 2-8 1999).

Foreign policy orientation towards either Russia or the West was also implicated in this debate. The supporters of the exhibition promoted Western orientation, which was seen as an alternative to the Russian influence. The opponents of the exhibition did not consciously try to propose Russian orientation. On the contrary, the presumably Russian origin of one of the American organizers, G. Guroff, was perceived as a problem. One of the opponents, the politician Nodar Natadze, stressed that "they say they have concluded an agreement with Grabov (sic). This Grabov is probably a Jew or a Baptist. As I was told, he used to work for the Soviet Ministry of Foreign Affairs, that is, he used to be a KGB agent" (Alia, 4-5 May 1999). However, the fact that numerous samples of Georgian material culture were still kept in Russian museums was never mentioned.

Another opposition was constructed along the lines of religious affiliation (Orthodoxy versus Catholicism and Protestantism) according to which the United States was seen as the enemy of the Orthodox world. The point of the link with the NATO campaign in Serbia was constantly made by the demonstrators, who interpreted the bombing of Belgrade negatively as "the attitude of the United States toward the Orthodox Nations." They also underlined that not only military objects but also the civilians and the cultural and architectural monuments were targeted in Serbia (Akhali Taoba, May 1 1999).

The exhibition thus created a context for exposing anti-Western feelings in the society at a time when their public expression was in opposition to the official state policies. For the extremist priest Father Basil (Mkalavishvili), the organizer of numerous attacks against minority religious groups, the exhibition was "an attempt on part of anti-Christian peoples to destroy the Georgian people intellectually" (Dilis Gazeti, May 11 1999; Akhali Taoba, April 30 1999). The students saw the United States as a country deprived of history; they denigrated "American imperialism" on 21st May, 1999, in a protest organized by the society Round Table in front of the American embassy, during which slogans displayed protest both against the issue of "the exporting of our national treasure" and the announcement of an increase in power prices by the American electricity supply company AES-Telavi (Dilis Gazeti, May 22 1999). 31 The opponents of the exhibition compared and contrasted the "priceless" value of the treasure and the sacrilegious character of the financial-political profit expected from the exhibition. 32 The link of the debate with the issue surrounding Georgian territory and the historical past was crucial. In the context in which non-Georgian influences were perceived as a threat, museum collections acquired the function of medieval icons: by force of their attachment to the territory of Georgia and its historical past they implied a perceived ability to protect the territory against foreign political, cultural and economic invasion. Those who saw the collections as holy objects, said that the bond between God and the nation could be broken if the icons left the territory. Paradoxically, the fact that the national treasury was saved from the Sovietization in the beginning of the 20th century by taking it abroad, i.e. leaving the territory of Georgia, was never mentioned. Father Basil (Mkalavishvili) suggested that those "who allowed that the grace granted by God could leave Georgia should be expelled from the Georgian nation" (Akhali Taoba, May 14 1999). Zurab Kiknadze, a distinguished folklorist, expert in religious studies and an editor of the Georgian translation of Max Weber's selected works in sociology of religion, explained that "to expose them abroad would be martyrdom of these icons" 31 Last but not least, a MacDonald's restaurant was opened in Tbilisi in June 1999, exacerbating the threat! 32 The opponents of the exhibition feared that it could be stolen, and that there was insufficient guarantee that the collections would remain a Georgian property. Another argument against the exhibition was related to a low amount of insurance, which was less than the selling price of a dress belonging to Elizabeth Taylor! Akhali Taoba, April 28 1999.

(Akhali 7 Dghe, April 30 -May 6 1999); a student feared that the grace of the icons "will be passed onto another country at a time when we need it" (Akhali Taoba, 8 May 1999). The Chief Director of the journal Resurrection of Lazar 33 justified his opposition to the exhibition by referring to the Ruis-Urbnisi Synod 34 and the Ecclesiastic Canon according to which if an icon was thrown on the ground after a war with a foreign aggressor, it would lose its sacred qualities and as a result, could not be taken back and used as an icon (Akhali Taoba, May 20 1999). This argument was also developed by Ilia II: he explained that in Orthodox countries like Russia or Georgia, the sanctity of the icons used to protect the towns from hardship and threats (Sakartvelos Resp'ublik'a, August 1999). The examples described above illustrate how religious attributes and symbols have been re-interpreted in a contemporary context, contributing to the processes of de-secularization.

CONCLUSION

The 19th century Russification of the Georgian Church and the 20th century forceful Soviet secularization of Georgian society contributed, like in other post-Soviet states, to the loss of the continuity and transmission of religious education and led to the greater diversity of the interpretation of religion, filling it with specific content (Capelle-Pogacean, Michel & Pace, 2008). The reshaping of Christianity in Georgia provided a background for new meanings and interpretations, and within the new political contexts, paved the way for its re-emergence as a central aspect of the national and political discourse. At the present religion appears to engage discourses and issues not directly related to it such as those of foreign policy orientations, relationships between the interests of different fractions of the political elite, corruption, national identity, ownership of the cultural heritage, etc.

Year 1999 appeared to be a turning point in the perception of Christianity and in its influence on the society. Religion started to be involved in all public political debates and as a result has become a constraint for all public figures and political actors, eventually contributing to the process of Georgia's de secularization. This process has had social and political implications. Since the "Rose revolution" in 2003 the tendency to give the religion a greater role has paradoxically grown: campaigns against religious minorities indeed stopped after the state put an end to their organizers' impunity; however, Mikheil Saakashvili's move to mobilize the resources provided by Orthodoxy for the use in his state policy and symbolism [START_REF]Formation Socio-Historique de la Nation Géorgienne: le legs des Identités Pré-modernes, les Idéologies et Acteurs Nationalistes[END_REF] demonstrates that a favorable relationship of the State with the Church and religion was now perceived as necessary for gaining the political legitimacy. The increased participation of the religion in public life and the intervention of the Church in political matters are illustrated by the proposition of the Catholicos Patriarch to establish a constitutional monarchy, which was backed by the opposition in the fall of 2007. Before that, on August 29 2007, the Catholicos Patriarch performed, for the first time in decades, a prayer and a service in front of the Anchiskhat'i icon at the Fine Arts museum, an illustration of the shifting borders between the secular and the religious spheres, between the political and the cultural implications of Christianity in Georgia.

ABOUT THE AUTHOR Silvia Serrano holds degrees in Slavic studies and French Literature. Her PhD was "Independent Georgia and its Caucasian Neighbours: identity issues and geopolitical restructuring." She lived in Russia and in Georgia from 1985 to 1990. She is maître de conférences (associate professor) in Political science at Clermont-Ferrand University (France) and research fellow at the Centre d'études des mondes russe, caucasien et centre-européen (Cercec, Paris). She currently works on the politics and identity (nationalism, minorities, and religion) in the Caucasus. She is an author of the book Géorgie. Sortie d'Empire, éditions du CNRS, Paris 2007.

  33 A semi-official religious periodical publication. The theme of Saint Lazar plays an important role in the Georgian religious national discourse. Drawing upon the work of the 10th century hymnograph, Ioane Zosime, entitled "Praise and Glorification of the Georgian Language" which claimed that Georgian is the language of the Judgment Day, this discourse relates the Resurrection of Lazar to the resurrection of the Georgian nation. 34 Ruis-Urbnisi diocese (in the Kartli province of eastern Georgia) is a major Ecclesiastic Council established by King David the Builder in 1103. It has supervised the vital issues of ecclesiastic as well as civil society in Georgia.

  14 In 1997, Georgian icons were exhibited in New York, see The Glory of Byzantium: Art and Culture of the

Middle Byzantine Era A.D. 843-1261, Metropolitan Museum of Art, NY, H.C. Evancs, W.D. Wixon. 15 See Sakartvelos Resp'ublik

'a, May 8 1999; K'viris P'alit'ra, August 2-3 1999.

This chapter is based on the systematic analyses of nine Georgian newspapers and press releases by the Patriarch of Georgia Ilia II from the April to November of 1999, as well as on interviews with the leaders of the protest, opposition leaders, public representatives, and museum staff. Student essays written in

were also added to the analysed data. I am grateful to Nino Kalandarishvili, Marina Elbakidze, Merab Tsindeliani and Emil Adelkhanov who helped with data collection. This study is a part of the broader research project on the Church and religious practice in contemporary Georgia supported and funded by the Maison des Sciences de l'Homme, and on the post-Soviet South, supported by the Agence Nationale pour la Recherche (ANR Les Suds).

All translations from Georgian are mine, unless stated otherwise.

The students were required to express their opinion about the following assertion as a selection criteria for enrolment in Master of Arts degree: "museum collections-pictures, old manuscripts as well as other cultural items will deteriorate if sent abroad and exposed in foreign museums" (based on the monitoring of 177 student essays for entering Master of Arts degree at the Ilia Chavchavadze University, Tbilisi in 2007.

Supporters of the former president, Zviad Gamsakhurdia, are known as "Zviadists" [lit., supporters of Zviad]. Although they had no MP in the Parliament, they represented a large part of the population. For many years after Zviad Gamsakhurdia was overthrown by coup d'état in 1991, the nation was divided into Zviadists and anti-Zviadists. Repercussions of this division are felt even today.

In 2004, he was sentenced to fifteen days of imprisonment for hooliganism after he tore down the posters of a provocative Polish artist at an exhibition at the National Parliamentary Library of Georgia. In 2006, he was briefly arrested again for insulting the Western-oriented rector of the Tbilisi State University. He was murdered in May 2007. See Civil Georgia http://www.civil.ge/eng/article.php?id=15150 Accessed on April 8 2008.

A prefect of the western Georgian city Samtredia during Gamsakhurdia rule, Zviad Dzidziguri is the leader of the Conservative Party. After Gamsakhurdia's expulsion Zviad Dzidziguri was sent to a colony, having been sentenced to 14 years of prison by the subsequent leadership. He was freed in 2000(Rezonansi, June 26 2000). Before the demonstration on 28 April, another protest action took part, demanding the liberation of the political prisoners-Zviadists, who were put in jail by Shevardnadze's government. From the very first days, there was confusion between the demonstrations organized by Sharadze and by Zviadists, because some of the demonstrators shifted from one protest action to the other.

During the Soviet rule Goguadze was a Deputy Rector of the Tbilisi State University in charge of ideology. As a former communist, he wrote an apologetic treatise about the Orthodox Christianity. Like G. Sharadze, V. Goguadze was awarded Order of Honor by E. Shevardnadze.

The Union of the Citizens of Georgia was a political party established by Shevardnadze after he took over leadership of Georgia in 1993. It became the majority group in Parliament in 2005, and remained the main political party until the "Rose revolution" in 2003.

In 2003, Sharadze joined the president-backed election alliance again and ran for MP in the Gldani constituency of the capital city Tbilisi.

The Moscow Patriarchy recognised autocephaly of the GOC in October 1943.

http://www.pravoslavie.ru/cgi-bin/sykon/client/display.pl?sid=381&did=2100, Accessed 5 April 2008.

The text was published in Sakartvelos Resp'ublik'a, October 15 2002. See also Keston Institute site, http://www.keston.org, accessed 5 April 2008, for the debates around the constitutional agreement.

Any object related to religion and belonging to the Church such as liturgical items, as well as relics (such as remains of the saints, for example) are referred to as religions items.

Dilis Gazeti, 1 May 1999, Axali 7 Dghe, April 30-May 6 1999, Akhali Taoba, May 1 1999.

In the late 1980s, for example, young volunteers restored churches and activists tried to protect the frescoes of David-Garedji monastery situated inside Russia's military polygon in K'akheti province.

Most of the survived and well-preserved medieval icons, such as Khakhuli or Anchiskhati, were supposed to give strength during battles against the enemy, and were seen as a protection. Other religious items, such as the Lailash Bible-a well known and rare Pentateuch [the first five books of the Old Testament] from the 10th century kept in Svaneti-are a testimony of how old and rich is the Georgian culture, hence contributing to its symbolic preservation. The list of items to be sent abroad for exhibition that was highlighted in the media was imprecise and changing. It was published in Akhali Taoba,April 30 1999 and May 1 1999, and K'viris P'alit'ra, April 26-May 2 1999.

Judith Dobrzynski, "Protests in Georgia Derail Art Show Headed for U.S." New York Times, August 4 1999.

See, for example, interview with the Georgian historian Mariam Lordkipanidze in Asaval-Dasavali, May 10-16 1999.

 29 See, for example, interview with Giorgi Antelava, the vice-director of The Institute of Oriental Studies in Tbilisi. Dilis Gazeti, May 1

1999.30 From 1978 to 1983, Ilia II was one of the presidents of the World Council of Churches.