DIFFERENTIAL STAINING OF NEURONS IN THE HUMAN RETINA WITH ANTIBODIES TO PROTEIN-KINASE-C ISOZYMES

Monoclonal antibodies to the three isozymes of protein kinase C (PKC) (alpha, beta, and gamma) were applied to postmortem human retina. Immunostaining was done on wholemount, or cryostat-sectioned retina, and visualized after ABC/DAB procedures by light (LM) and electron (EM) microscopy. The PCK-alpha antibody stained rod bipolar cells throughout the retina. EM analysis confirmed they were PKC-alpha-immunoreactive (IR) on their characteristic dendritic and axonal synaptology. Putative blue cone bipolar cells with wide-field axon terminals, stratifying in s5 of the inner plexiform layer (IPL), were also PKC-alpha-IR, and EM showed them to engage in narrow-cleft ribbon junctions in blue cone pedicles. The PKC-beta antibody stained cone bipolar cells, many amacrine cells, and most ganglion cells. Cone bipolar cells were stained all the way into the foveal center: both midget and diffuse varieties were included. The IPL was densely PKC-IR and individual neurons could not be identified on stratification patterns. EM of the outer plexiform layer (OPL) revealed that both flat and invaginating cone bipolar types were IR and that IR axon terminals were presynaptic in all strata of the IPL. The occurrence of PKC-beta-IR bipolar axons in stratum 2 of the IPL suggests that OFF-center as well as ON-center types were included. The PKC-gamma antibody gave inferior staining compared with results from the other two antibodies; however, two varieties of wide-field monostratified amacrine cell and a large-bodied ganglion cell type were discernible. PKC in one form or another appears to be a second messenger used in neurotransmission by both rod and cone systems and ON- and OFF-center systems in the human retina.