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Volume 17, Number 9, Issue of May 1, 1997 pp. 3014-3023
Copyright ©1997 Society for Neuroscience

The mGluR6 5 Upstream Transgene Sequence Directs a Cell-Specific and Developmentally Regulated Expression in Retinal Rod and ON-Type Cone Bipolar Cells

Received Dec. 20, 1996; revised Feb. 7, 1997; accepted Feb. 11, 1997.

Yoshiki Ueda¹, Hideki Iwakabe¹, Masayuki Masu¹, Misao Suzuki², and Shigetada Nakanishi¹

¹ Department of Biological Sciences, Kyoto University Faculty of Medicine, Kyoto 606-01, Japan, and ² Institute of Molecular Embryology and Genetics, Kumamoto University Medical School, Kumamoto 862, Japan

We generated transgenic mice, using 9.5 kilobase pairs of the 5 upstream sequence from the mouse metabotropic glutamate receptor subtype 6 (mGluR6) gene fused to the -galactosidase (lacZ) reporter gene, and investigated the promoter function of the cell-specific and developmentally regulated expression of mGluR6. Most of the independent transgenic lines commonly showed the lacZ expression in the defined cell layers of the retina, and four transgenic lines were characterized in detail for cell-specific lacZ expression patterns by X-gal staining and lacZ immunostaining. The lacZ-expressing retinal cells were classified into two cell types. One cell type was identified as rod bipolar cells on the basis of colocalization of protein kinase C (PKC) immunoreactivity and morphological criteria. The other cell type was PKC-immunonegative and resided at the cell layers corresponding precisely to ON-type cone bipolar cells. The latter bipolar cells were found to exist as a large cell population comparable to rod bipolar cells. This observation was confirmed by coimmunostaining of dissociated retinal cells with the lacZ and PKC antibodies. The ontogeny analysis indicated that the lacZ expression completely agrees with a temporal expression pattern of mGluR6 during retinal development. This study demonstrates that the mGluR6 5 upstream genomic sequence is capable of directing a cell-specific and developmentally regulated expression of mGluR6 in ON-type bipolar cells and supports the view that mGluR6 is responsible for ON responses in both the rod and cone systems.

Key words: transgenic mouse; metabotropic glutamate receptor; -galactosidase; retinal bipolar cells; immunostaining

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