Yoshida K, Imaki J, Koyama Y, Harada T, Shinmei Y, Oishi C, Matsushima-Hibiya Y, Matsuda A, Nishi S, Matsuda H, Sakai M
Department of Ophthalmology, Hokkaido University School of Medicine, Sapporo, Japan.
Invest Ophthalmol Vis Sci. 1997 Nov;38(12):2679-83.
To examine the expression of maf-1 and maf-2 protocogenes in the developing rat lens.
Maf-1 and maf-2 transcripts were assayed in rat lenses on embryonic days 13 and 16 (E13 and E16) by in situ hybridization using single-stranded RNA probes. Proteins encoded by the maf-2 gene were assayed immunocytochemically in embryonic (E12, 13, 16, 19) and postnatal day 14 and 90 (P14 and P90) lenses.
In embryonic lenses, we detected maf-1 messenger RNA (mRNA) in the lens epithelium and maf-2 mRNA diffusely distributed in the lens fiber cells. By immunocytochemistry, Maf-2 was detected on E12 in the nuclei of almost all lens pit cells. On days E13, E16, and E19, however, lens epithelial cells showed no immunoreactivity, but nuclei of fiber cells reacted strongly. On P14, nuclei containing Maf-2 protein were confined to the equator of the lens, but at 3 months of age, no Maf-2 could be detected in the rat lens. Western blotting showed that the anti-Maf-2 antiserum reacted with a single protein, of molecular weight approximately 39 kDa, in rat lens.
Results showed the spatial and temporal regulation of maf gene expression and suggest that these genes participate in transcriptional regulation during the development of the lens in the rat.
研究原癌基因maf - 1和maf - 2在发育中的大鼠晶状体中的表达情况。
采用单链RNA探针原位杂交技术，检测胚胎第13天和16天（E13和E16）大鼠晶状体中的maf - 1和maf - 2转录本。利用免疫细胞化学方法检测胚胎期（E12、13、16、19）以及出生后第14天和90天（P14和P90）晶状体中maf - 2基因编码的蛋白质。
在胚胎晶状体中，我们在晶状体上皮细胞中检测到maf - 1信使核糖核酸（mRNA），而maf - 2 mRNA在晶状体纤维细胞中呈弥漫性分布。通过免疫细胞化学检测发现，在E12时，几乎所有晶状体窝细胞的细胞核中都能检测到Maf - 2。然而，在E13、E16和E19时，晶状体上皮细胞未显示免疫反应性，但纤维细胞的细胞核反应强烈。在P14时，含有Maf - 2蛋白的细胞核局限于晶状体赤道部，但在3月龄时，大鼠晶状体中未检测到Maf - 2。蛋白质免疫印迹分析表明，抗Maf - 2抗血清与大鼠晶状体中一种分子量约为39 kDa的单一蛋白质发生反应。
结果显示了maf基因表达的时空调控，提示这些基因参与大鼠晶状体发育过程中的转录调控。
