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 microm 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.