We present here a reproducible and economical protocol for the simultaneous detection of glutamate decarboxylase (GDC) – the key enzyme in γ-aminobutyric acid (GABA) synthesis – and synaptophysin – a protein maker for synaptic vesicles – using confocal laser microscopy. In the rat cerebellar cortex, GDC labels Purkinje cells and the pinceaux of nerve fibers in their basal parts, and was unevenly distributed in the neuropil of the molecular and granular layers. Synaptophysin in the molecular layer clearly outlined the large dendrites of Purkinje cells and in the granular layer it labeled some cerebellar glomeruli, i.e., mossy fiber terminals. GDC-immunopositive structures (GABAergic axons of stellate neurons, i.e., Golgi cells) were often located on the peripheries of glomeruli. The peripheral parts of glomeruli showed colocalization of GDC- and synaptophysin-immunopositive structures, which is evidence for the presence of GABAergic synapses in this area.
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References
E. G. Gilerovich, “Immunohistochemical studies of the structural bases of inhibition in the central nuclei of the cerebellum in mice,” Ros. Fiziol. Zh., 84, No. 12, 1325–1332 (1998).
E. G. Gilerovich, T. R. Moshonkina, E. A. Fedorova, et al., “Morphofunctional characteristics of the lumbar enlargement of the rat spinal cord,” Morfologiya, 132, No. 5, 33–37 (2007).
A. V. Gilyarov, O. V. Kirik, and D. E. Korzhevskii, “Comparative analysis of methodological approaches used for simultaneous detection of multiple antigens in immunohistochemical studies,” Morfologiya, 138, No. 5, 59–64 (2010).
D. E. Korzhevskii, O. V. Kirik, M. N. Karpenko et al., Theoretical Bases and Practical Application of Immunocytochemical Methods, SpetsLit, St. Petersburg (2014).
D. E. Korzhevskii,, E. G. Sukhorukova, E. G. Gilerovich, et al.,“Advantages and disadvantages of zinc-ethanol-formaldehyde as a fixative for immunocytochemical studies and confocal laser microscopy,” Morfologiya, 143, No. 2, 81–85 (2013).
B. S. Sukhareva, E. L. Darii, and R. R. Khristoforov, “Glutamate decarboxylase: structure and catalytic properties,” Usp. Biol. Khim., 41, 131–162 (2001).
G. J. Evans and M. A. Cousin, “Tyrosine phosphorylation of synaptophysin in synaptic vesicle recycling,” Biochem. Soc. Trans., 33, No. 6, 1350–1353 (2005).
N. Garin and G. Escher, “The development of inhibitory synaptic specializations in mouse deep cerebellar nuclei,” Neuroscience, 105, 431–441 (2001).
M. Manto and C. I. De Zeeuw, “Diversity and complexity of roles of granule cells in the cerebellar cortex,” Cerebellum, 11, 1–4 (2012).
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Translated from Morfologiya, Vol. 147, No. 1, pp. 74–77, January–February, 2015.
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Korzhevskii, D.E., Gilerovich, E.G., Kirik, O.V. et al. Simultaneous Detection of Glutamate Decarboxylase and Synaptophysin in Paraffin Sections of the Rat Cerebellum. Neurosci Behav Physi 46, 106–109 (2016). https://doi.org/10.1007/s11055-015-0205-6
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DOI: https://doi.org/10.1007/s11055-015-0205-6