Isolation of ON Bipolar Cell Genes via hrGFP-coupled Cell Enrichment Using the mGluR6 Promoter

• DOI: 10.1093/jb/mvp038
• Publication Date: 2009-03-07T01:54:13Z
• Authors: Yoshiaki Nakajima; Masaki Moriyama; Masakazu Hattori; Nagahiro Minato; Shigetada Nakanishi
• Keywords: 0301 basic medicine; Retinal Bipolar Cells; Calcium Channels, L-Type; Reverse Transcriptase Polymerase Chain Reaction; Green Fluorescent Proteins; TRPM Cation Channels; Mice, Transgenic; Receptors, Metabotropic Glutamate; Immunohistochemistry; Mice, Inbred C57BL; Mice; 03 medical and health sciences; Animals; Humans; Female; Promoter Regions, Genetic; In Situ Hybridization; RGS Proteins; Oligonucleotide Array Sequence Analysis

mGluR6 expression is a characteristic property of retinal ON bipolar cells. mGluR6 is also the causal gene for a form of congenital night blindness. To elucidate physiological and pathological functions of ON bipolar cells and mGluR6, we thought it important to identify genes specifically expressed in them. We thus made transgenic mouse lines expressing humanized Renilla reniformis green fluorescent protein (hrGFP), under the control of the mGluR6 promoter. From their retina, we isolated hrGFP-positive cells by cell sorting, and analysed the gene-expression profile of these cells by using DNA microarray. Further analysis revealed that about half of the initially selected ON bipolar cell genes were expressed in the expected retinal layer. We confirmed previously ambiguous retinal localization of regulator of G-protein signalling 11 (RGS11) and transient receptor potential cation channel, subfamily M, member 1 (TRPM1). In addition, we showed the expression of calcium channel, voltage-dependent, alpha2/delta subunit 3 (Cacna2d3) in ON bipolar cells for the first time. Although we could not completely exclude the possibility that a small population of hrGFP-positive cells might not be ON bipolar cells, these mice as well as our strategy would be highly valuable for the further analysis of ON bipolar cells.