生物
原位杂交
视网膜
分子生物学
免疫印迹
北方斑点
恒河猴
免疫组织化学
免疫细胞化学
神经节细胞层
信使核糖核酸
基因
遗传学
免疫学
内分泌学
神经科学
作者
Takashi Kitaoka,Amy E. Aotaki-Keen,Leonard M. Hjelmeland
出处
期刊:PubMed
[National Institutes of Health]
日期:1994-07-01
卷期号:35 (8): 3189-98
被引量:33
摘要
PURPOSE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
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