Excessive calcium is thought to be a critical step in various neurodegenerative processes including ischemia. Calbindin D28k (CB), calretinin (CR), and parvalbumin (PV), members of the EF-hand calcium-binding protein family, are thought to play a neuroprotective role in various pathologic conditions by serving as a buffer against excessive calcium. The expression of CB, PV and CR in the ischemic rat retina induced by increasing intraocular pressure was investigated at the transcript and protein levels, by means of the quantitative real-time reverse transcription-polymerase chain reaction, western blot and immunohistochemistry. The transcript and protein levels of CB, which is strongly expressed in the horizontal cells in both normal and affected retinas, were not changed significantly and the number of CB-expressing horizontal cells remained unchanged throughout the experimental period 8 weeks after ischemia/reperfusion injury. At both the transcript and protein levels, however, CR, which is strongly expressed in several types of amacrine, ganglion, and displaced amacrine cells in both normal and affected retinas, was decreased. CR-expressing ganglion cell number was particularly decreased in ischemic retinas. Similar to the CR, PV transcript and protein levels, and PV-expressing AII amacrine cell number were decreased. Interestingly, in ischemic retinas PV was transiently expressed in putative cone bipolar cell types possibly those that connect with AII amacrine cells via gap junctions. These results suggest that these three calcium binding proteins may play different neuroprotective roles in ischemic insult by their ability to buffer calcium in the rat retina.
Amacrine Cells ; Animals ; Blotting, Western ; Calcium ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Cell Count ; Ganglion Cysts ; Gap Junctions ; Humans ; Immunohistochemistry ; Intraocular Pressure ; Ischemia ; Neurons ; Proteins ; Rats ; Retina

Horizontal cells (HCs) of the mammalian retina are interneurons that provide negative feedback to photoreceptors in the outer plexiform layer (OPL) where the first synapse occurs and contribute to the center surround antagonism that underlies the receptive field properties of many retinal neurons. These functions of HCs are thought to be attributed to their coupled network via gap junctions. Two kinds of connexin (Cx) proteins, Cx50 and 57 are known to form gap junctions of HCs. However, little is known about precise localization of gap junctions within HCs. Thus, this study was designed to determine the localization of HC gap junctions at subcellular level. In vertical ultrathin sections of the rabbit retina, gap junctions composed of Cx50 and 57 were identified in the OPL by the electron-dense reaction products. Each Cx50 and 57 gap junction on putative HC processes showed its own distinct features. Cx50 gap junction was bigger in size and localized more proximally than Cx57. In addition, Cx57 gap junctions had distinct shape. That is, about a half of them appeared to be invaginated or endocytosed in shape. The differences in shape, size and subcellular localization between Cx50 and 57 gap junctions may provide the insights into the function of different types of horizontal cell.
Gap Junctions ; Interneurons ; Proteins ; Retina ; Retinal Neurons ; Synapses