Hernandez E R, Hurwitz A, Botero L, Ricciarelli E, Werner H, Roberts C T, LeRoith D, Adashi E Y
Department of Obstetrics and Gynecology, University of Maryland School of Medicine, Baltimore 21201.
Mol Endocrinol. 1991 Dec;5(12):1799-805. doi: 10.1210/mend-5-12-1799.
The intraovarian insulin-like growth factor (IGF) system constitutes a triad composed of ligands, receptors, and binding proteins. Although conventional radioligand receptor assays have documented the presence of specific receptors for insulin and insulin-like peptides in some rat somatic ovarian cell types, the exact cellular localization and hormonal regulation of the receptors in question remain matters of inquiry. To reevaluate the very presence, cellular localization, and hormonal regulation of the IGF receptor gene family in the rat ovary, solution hybridization/RNase protection assays were used wherein ovarian total RNA (20 micrograms) from immature (21-23 days old) rats was hybridized with 32P-labeled type I IGF receptor, type II IGF/mannose-6-phosphate receptor, and insulin receptor riboprobes. Single protected fragments 261 (type I IGF receptor), 500 (type II IGF/mannose-6-phosphate receptor), and 478 (insulin receptor) bases long were evident in whole ovary, granulosa, and theca-interstitial cells. Hypophysectomy of immature rats led to significant (P less than 0.05) albeit variable decrements in the relative (densitometrically quantified) ovarian abundance of transcripts corresponding to the type I IGF (but not insulin or type II IGF/mannose-6-phosphate) receptor. Treatment of immature hypophysectomized rats with FSH (10 micrograms/rat.day x 2.5 days) resulted in a significant (P less than 0.05) increase (4-fold) in transcripts corresponding to the type I IGF receptor in both whole ovarian material and freshly isolated granulosa cells. Similar (3.7-fold) increments (P less than 0.05) were noted after treatment with a diethylstilbestrol-containing sc silastic implant applied for a total of 5 days.(ABSTRACT TRUNCATED AT 250 WORDS)
卵巢内胰岛素样生长因子(IGF)系统由配体、受体和结合蛋白组成三联体。尽管传统的放射性配体受体分析已证明在某些大鼠卵巢体细胞类型中存在胰岛素和胰岛素样肽的特异性受体,但相关受体的确切细胞定位和激素调节仍有待研究。为了重新评估大鼠卵巢中IGF受体基因家族的存在、细胞定位和激素调节,采用了溶液杂交/RNase保护分析,将来自未成熟(21 - 23日龄)大鼠的卵巢总RNA(20微克)与32P标记的I型IGF受体、II型IGF/甘露糖-6-磷酸受体和胰岛素受体核糖探针杂交。在整个卵巢、颗粒细胞和卵泡膜-间质细胞中明显可见长度分别为261个碱基(I型IGF受体)、500个碱基(II型IGF/甘露糖-6-磷酸受体)和478个碱基(胰岛素受体)的单一受保护片段。未成熟大鼠垂体切除导致对应I型IGF(而非胰岛素或II型IGF/甘露糖-6-磷酸)受体的转录本相对(经光密度定量)卵巢丰度显著(P < 0.05)下降,尽管下降幅度有所不同。用FSH(10微克/大鼠·天×2.5天)治疗未成熟垂体切除大鼠,导致整个卵巢组织和新鲜分离的颗粒细胞中对应I型IGF受体的转录本显著(P < 0.05)增加(4倍)。在用含己烯雌酚的皮下硅橡胶植入物治疗5天后,也观察到类似的(3.7倍)增加(P < 0.05)。(摘要截短于第250字)