Kitaoka T, Aotaki-Keen A E, Hjelmeland L M
Department of Ophthalmology, University of California, Davis 95616.
Invest Ophthalmol Vis Sci. 1994 Jul;35(8):3189-98.
To determine the distribution of mRNA transcribed from the FGF-5 gene and the distribution of FGF-5 protein in the normal adult retina of the rhesus macaque.
Freshly enucleated globes from rhesus macaque were prepared for Northern blot analysis, in situ hybridization, and immunohistochemical studies. A 350 base pair sequence from the human FGF-5 was subcloned and used to prepare 35S-labeled cRNAs for Northern and in situ experiments. Antipeptide antibodies were raised against an aminoterminal sequence as well as an internal sequence and were affinity purified on recombinant FGF-5 coupled to AffiGel-10. The specificity of antibody probes was verified by dot blot and Western blot analyses.
Ganglion cells and photoreceptors express the highest levels of FGF-5 mRNA, although all neurons and the retinal pigment epithelium (RPE) were clearly labeled. Immunohistochemical staining similarly revealed a widespread distribution of the protein, with prominent labeling observed in photoreceptor inner segments, especially in cone cells. Plexiform layers and RPE cells were also clearly labeled.
Clear evidence was established for the expression of FGF-5 in the retina, where it may play an important extracellular role as a secreted member of the FGF gene family.
