Expression of retinol-binding protein and cellular retinol-binding protein in the bovine ovary
Expression of retinol-binding protein and cellular retinol-binding protein in the bovine ovary
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DOI：
10.1002/mrd.10225
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发表时间：
2003-03-01
影响因子：
2.5
通讯作者：
Godkin, JD
中科院分区：
文献类型：
--
作者：
Brown, JA;Eberhardt, DM;Godkin, JD
视黄醇（维生素A）是生殖所必需的，类维生素A被认为在卵巢类固醇生成、卵母细胞成熟和早期胚胎发育中发挥作用。视黄醇通过视黄醇结合蛋白（RBP）进行全身和细胞间转运。在细胞内，细胞视黄醇结合蛋白（CRBP）在视黄醇积累和代谢中起作用。由于类维生素A的作用部分由类维生素A结合蛋白介导，因此本研究的目的是研究牛卵巢中RBP和CRBP的细胞特异性表达。免疫细胞化学分析（ICC）将RBP定位于窦和窦前卵泡的卵泡膜和颗粒细胞层，在大的健康卵泡的细胞中染色最强烈。动脉血管外膜的图尼卡膜也显示RBP染色。CRBP的免疫染色在腔前卵泡的颗粒细胞中最强，在有腔卵泡的卵泡膜和颗粒细胞中存在，但减弱。在黄体内，这两种蛋白质在大的黄体细胞中观察到，但只有RBP在小的黄体细胞中观察到。北方杂交结果显示，有腔卵泡的卵泡膜细胞、颗粒细胞和黄体组织中均有RBP和CRBP mRNA的表达。从培养细胞或外植体的培养基中放射性标记的RBP的免疫复合物沉淀，随后通过SDS-PAGE和荧光照相术证明了卵泡膜细胞、颗粒细胞和黄体细胞的RBP合成和分泌。收集卵泡液，从小（
Retinol (vitamin A) is essential for reproduction, and retinoids have been suggested to play a role in ovarian steroidogenesis, oocyte maturation, and early embryonic development. Retinol is transported systemically and intercellularly by retinol-binding protein (RBP). Within the cell, cellular retinol-binding protein (CRBP) functions in retinol accumulation and metabolism. Since the actions of retinoids are mediated, in part, by retinoid-binding proteins, the objective of this study was to investigate cell-specific expression of RBP and CRBP in the bovine ovary. Immunocytochemical analysis (ICC) localized RBP to the thecal and granulosa cell layers of antral and preantral follicles with the most intense staining in the cells of large, healthy follicles. The tunica adventitia of arterial blood vessels also exhibited RBP staining. Immunostaining of CRBP was most intense in the granulosa cells of preantral follicles and present, but diminished, in thecal and granulosa cells of antral follicles. Within the corpus luteum, both proteins were observed in large luteal cells, but only RBP was observed in small luteal cells. Northern blot analysis demonstrated that thecal and granulosa cells from antral follicles and luteal tissue expressed RBP and CRBP mRNA. Synthesis and secretion of RBP by thecal cells, granulosa cells, and luteal cells were demonstrated by immune-complex precipitation of radiolabeled RBP from the medium of cultured cells or explants, followed by SDS-PAGE and fluorography. Follicular fluid was collected from small (