A Thy1-CFP DBA/2J mouse line with cyan fluorescent protein expression in retinal ganglion cells.
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.
Raymond
Iona D
ID
Department of Neurobiology, David Geffen UCLA School of Medicine, Los Angeles, CA 90095-1763, USA. iona.d.raymond@ucla.edu
Pool
Angela L
AL
Vila
Alejandro
A
Brecha
Nicholas C
NC
D025042:Amacrine Cells / Q000378:metabolism / Q000473:pathology; D000818:Animals; D001369:Axons / Q000378:metabolism / Q000473:pathology; D001947:Breeding; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D005902:Glaucoma, Open-Angle / Q000378:metabolism / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D009414:Nerve Growth Factors / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012165:Retinal Ganglion Cells / Q000378:metabolism* / Q000473:pathology; D050827:Syntaxin 1 / Q000378:metabolism; D018800:Thy-1 Antigens / Q000235:genetics*; D050819:Transcription Factor Brn-3A / Q000378:metabolism
C072167:CAT development factor; C485182:Cyan Fluorescent Protein; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D009687:Nuclear Proteins; C494189:Pou4f1 protein, mouse; D011993:Recombinant Fusion Proteins; C494214:Stx1a protein, mouse; D050827:Syntaxin 1; D018800:Thy-1 Antigens; D050819:Transcription Factor Brn-3A; D049452:Green Fluorescent Proteins