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A Thy1-CFP DBA/2J mouse line with cyan fluorescent protein expression in retinal ganglion cells

  • IONA D. RAYMOND (a1), ANGELA L. POOL (a1), ALEJANDRO VILA (a2) and NICHOLAS C. BRECHA (a1) (a3) (a4) (a5)

Abstract

A DBA/2J (D2) transgenic mouse line with cyan fluorescent protein (CFP) reporter expression in ganglion cells was developed for the analysis of ganglion cells during progressive glaucoma. The Thy1-CFP D2 (CFP-D2) line was created by congenically breeding the D2 line, which develops pigmentary glaucoma, and the Thy1-CFP line, which expresses CFP in ganglion cells. Microsatellite marker analysis of CFP-D2 progeny verified the genetic inclusion of the D2 isa and ipd loci. Specific mutations within these loci lead to dysfunctional melanosomal proteins and glaucomatous phenotype in D2 mice. Polymerase chain reaction analysis confirmed the inclusion of the Thy1-CFP transgene. CFP-fluorescent ganglion cells, 6–20 μm in diameter, were distributed in all retinal regions, CFP processes were throughout the inner plexiform layer, and CFP-fluorescent axons were in the fiber layer and optic nerve head. Immunohistochemistry with antibodies to ganglion cell markers NF-L, NeuN, Brn3a, and SMI32 was used to confirm CFP expression in ganglion cells. Immunohistochemistry with antibodies to amacrine cell markers HPC-1 and ChAT was used to confirm weak CFP expression in cholinergic amacrine cells. CFP-D2 mice developed a glaucomatous phenotype, including iris disease, ganglion cell loss, attrition of the fiber layer, and elevated intraocular pressure. A CFP-D2 transgenic line with CFP-expressing ganglion cells was developed, which has (1) a predominantly D2 genetic background, (2) CFP-expressing ganglion cells, and (3) age-related progressive glaucoma. This line will be of value for experimental studies investigating ganglion cells and their axons in vivo and in vitro during the progressive development of glaucoma.

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Corresponding author

*Address correspondence and reprint requests to: Iona D. Raymond, Department of Neurobiology, David Geffen School of Medicine at UCLA, 10833 Le Conte Avenue, Box 951763, Los Angeles, CA 90095-1763. E-mail: iona.d.raymond@ucla.edu

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A Thy1-CFP DBA/2J mouse line with cyan fluorescent protein expression in retinal ganglion cells

  • IONA D. RAYMOND (a1), ANGELA L. POOL (a1), ALEJANDRO VILA (a2) and NICHOLAS C. BRECHA (a1) (a3) (a4) (a5)

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