GABA and glycine in retinal amacrine cells: combined Golgi impregnation and immunocytochemistry

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Abstract
Golgi-impregnated amacrine cells in the all-cone lizard retina ( Anolis carolinensis ) were characterized on the bases of dendritic and somatic criteria. Four major cell categories, comprising 23 types were identified: three non-stratified, 13 monostratified, five bistratified, and two tristratified types. Four of the cell types comprised two to four subtypes based on stratification of their dendrites within the inner plexiform layer (IPL). Golgi impregnation strongly favoured monostratified amacrine cells with cell bodies at the proximal margin of the inner nuclear layer. The neurotransmitter content of each of the 23 amacrine cell types was examined by combined Golgi-immunocytochemistry after morphological classification. Putative neurotransmitters examined included gamma-aminobutyric acid (GABA), glycine (GLY) and aspartate (ASP). Seventeen cell types showed GABA-immunoreactivity (IR), three cell types showed GLY-IR, and four cell types showed neither GABA-IR nor GLY-IR. No cell types showed ASP-IR. Each cell type had a characteristic neurochemical signature, with the exception of one monostratified cell type that showed three different neurochemical signatures. Postembedding immunocytochemistry on conventionally processed retinas confirmed the localization of glutamic acid decarboxylase, the synthetic enzyme for GABA, to cells similar to several of the GABA-IR Golgi-stained types. Postembedding immunocytochemistry for tyrosine hydroxylase (the synthetic enzyme for catecholamines) and GABA on serial sections demonstrated colocalization of GABA and a catecholamine,probably dopamine, in a bistratified amacrine cell type. We conclude that GABA-IR amacrine cell types are more numerous and morphologically heterogeneous than GLY-IR amacrine cells. The morphological heterogeneity and, with one exception, exclusivity of GABA-IR and GLY-IR amacrine cell types indicate that both neurotransmitters play a variety and different functional roles in the lizard inner retina.