Immunocytochemical localization of the GABAc receptor rho subunits in the mammalian retina.

Polyclonal antibodies against the N terminus of the rat rho 1 subunit were generated to study the distribution of GABAc receptors in the mammalian retina. The specificity of the antibodies was tested in Western blots and transfected HEK-293 cells. No cross-reactivity with the GABAA receptor subunits alpha 1-3, beta 1-3, gamma 2, delta or with the glycine receptor subunits alpha 1 and beta could be detected. In contrast, the rho 1, rho 2, and rho 3 subunits were all recognized by the antibodies. In vertical sections of rat, rabbit, cat, and macaque monkey retinae, strong punctate immunoreactivity was present in the inner plexiform layer. Weaker immunoreactivity was also present in the outer-plexiform layer, and cell bodies of bipolar cells were faintly labeled. Double immunostaining of vertical sections and immunostaining of dissociated rat retinae showed the punctate immunofluorescence to colocalize with bipolar cell axon terminals. The puncta possibly represent clustering of the rho subunits at postsynaptic sites.