Gene expression profile of the adult human retinal ganglion celllayer

[摘要] Purpose: Pathophysiological events in the retinal ganglion celllayer (GCL) are a prominent feature of several optic neuropathiesincluding glaucoma. The purpose of this study was to identify andcatalog genes whose expression in the human retina is restricted to theGCL.Methods: Laser capture microdissection (LCM) technology was used toisolate tissue from the perimacular retina of three human donors withoutretinal or optic nerve disease. RNA was isolated from the (1) retinalGCL and (2) the inner and outer nuclear layers of the same retina, andthe gene expression profiles of both fractions were determined usingAffymetrix Hu133Plus 2.0 GeneChips. Data were analyzed to identify thosegenes whose expression is substantially more prevalent in the GCL whencompared to the outer retinal layers. Differential expression ofselected genes was confirmed by real-time PCR and immunohistochemistry.Results: The results show that mRNA levels of previously describedganglion cell markers, e.g. the neurofilament genes (NEFH, NEF3, orNEFL) and the Brn3a transcription factor (POU4F1), were highly enrichedin the isolated GCL fraction. In contrast, transcripts for genesassociated with phototransduction (RHO), photoreceptor development(NR2E3), or interphotoreceptor matrix constituents (IMPG1) were nearlyabsent from the GCL fraction. Using bioinformatics approaches over 80genes were identified whose expression in the human retina appears to belimited to the GCL.Conclusions: We have successfully used LCM technology to generategene expression profiles of highly enriched GCL fractions of the normalhuman retina. These data not only provide clues to the normal functionof retinal ganglion cells but also serve as a resource in thedevelopment of ganglion cell specific markers or transfection vectors,and the identification of candidate genes for hereditary forms ofglaucoma.

[学科分类] 生物化学/生物物理