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Journal of Biochemistry Advance Access published online on March 6, 2009
Journal of Biochemistry, doi:10.1093/jb/mvp038
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© The authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

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

Yoshiaki Nakajima1, Masaki Moriyama2, Masakazu Hattori2,3, Nagahiro Minato2 and Shigetada Nakanishi4

From 1Department of Biological Sciences, Faculty of Medicine, Kyoto University, Kyoto, 606-8501, Japan; 2Department of Immunology and Cell Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan; 3Department of Biosciences, School of Science, Kitasato University, Sagamihara, Kanagawa, 228-8555, Japan; and 4Department of Systems Biology, Osaka Bioscience Institute, Suita, Osaka 565-0874, Japan.

Corresponding Author: Yoshiaki Nakajima Department of Biological Sciences, Faculty of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, Japan, 606-8501 telephone: 81-75-753-4407 fax: 81-75-753-4404 e-mail: yoshiaki{at}phy.med.kyoto-u.ac.jp

Received January 8, 2009; Accepted February 26, 2009


  Abstract

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 analyzed 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 signaling 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.

Key Words: bipolar cells, Cacna2d3, mGluR6, RGS11, TRPM1


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