Previous results from our genetic analyses using pedigrees from a French Canadian population suggested that the interval delimited by markers on chromosome 12, D12S86 and D12S378, was the most probable genomic region to contain a susceptibility gene for affective disorders. Association studies with microsatellite markers using a case/control sample from the same population (n = 427) revealed significant allelic associations between the bipolar phenotype and marker NBG6. Since this marker is located in intron 9 of the P2RX7 gene, we analyzed the surrounding genomic region for the presence of polymorphisms in regulatory, coding and intron/exon junction sequences. Twenty four (24) SNPs were genotyped in a case/control sample and 12 SNPs in all pedigrees used for linkage analysis. Allelic, genotypic or family-based association studies suggest the presence of two susceptibility loci, the P2RX7 and CaMKK2 genes. The strongest association was observed in bipolar families at the non-synonymous SNP P2RX7-E13A (rs2230912, P-value = 0.000708), which results from an over-transmission of the mutant G-allele to affected offspring. This Gln460Arg polymorphism occurs at an amino acid that is conserved between humans and rodents and is located in the C-terminal domain of the P2X7 receptor, known to be essential for normal P2RX7 function.

Uptake of GABA was demonstrated in rat brain synaptic vesicles which were prepared by a new and efficient procedure. The uptake activity co‐purified with the synaptic vesicles during the isolation procedure. The purity of the vesicle fraction was rigorously examined by analysis of marker enzymes and marker proteins and also by immunogold electron microscopy using antibodies against p38 (synaptophysin). Contamination by other cellular components was negligible, indicating that GABA uptake by the synaptic vesicle fraction is specific for synaptic vesicles and not due to the presence of other structure possessing GABA uptake or binding activities. GABA uptake was ATP dependent and similar to the uptake of glutamate, which was assayed for a comparison. Both uptake activities were independent of sodium. They were inhibited by the uncoupler carbonyl cyanide 4‐(trifluoromethoxy)phenylhydrazone, indicating that the energy for the uptake is provided by an electrochemical proton gradient. This gradient is generated by a proton ATPase of the vacuolar type as suggested by the effects of various ATPase inhibitors on neurotransmitter uptake and proton pumping. Competition experiments revealed that the transporters for GABA and glutamate are selective for the respective neurotransmitters.

Cholinergic synaptic vesicles from the electric organ of Torpedo marmorata have been purified to a constant composition and a higher transmitter content than previously reported. By optimising the extraction conditions and using a two‐step purification on discontinuous and continuous sucrose density gradients, 10‐fold higher acetylcholine and ATP values per weight of protein were obtained. The purity of the vesicle preparation was confirmed by electronmicroscopy, absence of marker enzymes, behaviour in density gradient centrifugation, as well as by a specific and reproducible protein composition. Vesicles contain 6.9 μmol acetylcholine and 1.0 μmol ATP per mg protein. The lipid/protein ratio of 3.5 (w/w) indicates a lipid‐rich membrane. The value suggests the absence of a proteinaceous core. Upon dodecylsulphate gel electrophoresis a distinct protein pattern is obtained with components ranging from 20000 to 160000 in molecular weight. Vesiculin, reported earlier to be a low molecular‐weight vesicle protein, is not detected. One of the major bands comigrates with muscle actin from the same animal. Further characterisation of this protein by two‐dimensional gel electrophoresis suggested that it is an actin‐like polypeptide. Evidence for a specific association of this actin‐like protein with vesicles and its possible involvement in the neurosecretory process is discussed.

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Plasma membranes were purified from purely cholinergic nerve endings (synaptosomes) isolated from the electric organ of Torpedo marmorata. Synaptosomes were lysed, membranes recovered and further separated by density gradient centrifugation. A fraction was obtained enriched in 5'-nucleotidase, Na' , K+-activated ATPase and acetylcholine esterase. Morphological examination showed abundant membrane fragments of the size range of synaptosomes and few of vesicle size. The fraction has a characteristic protein composition upon gel electrophoresis. Five reproducible major bands with apparent M