Liu J, Koenigsfeld A T, Cantley T C, Boyd C K, Kobayashi Y, Lucy M C
Department of Animal Sciences, University of Missouri, Columbia, Missouri 65211, USA.
Biol Reprod. 2000 Sep;63(3):942-52. doi: 10.1095/biolreprod63.3.942.
Ovarian follicular growth and steroidogenesis are controlled by the interaction of insulin-like growth factors (IGFs) and gonadotropins. The objective was to determine the temporal and spatial relationships for gonadotropin receptor, steroidogenic enzyme, and IGF system gene expression during the development of preovulatory porcine follicles. Sows (n = 18) were weaned and follicles were monitored by transrectal ultrasonography. Ovaries were collected from sows when the mean diameter of the preovulatory follicular cohort was approximately 2, 4, 6, or 8 mm. mRNA were measured by in situ hybridization for individual follicles within the preovulatory cohort (3 to 5 follicles per sow). Patterns of gene expression detected by in situ hybridization were confirmed by RNase protection analyses of pooled RNA samples. The amount of LH receptor mRNA and steroidogenic enzyme mRNA (17alpha-hydroxylase and aromatase) increased as the mean diameter of the follicular cohort increased from 2 to 6 mm, but then decreased abruptly for 8-mm follicles. Estradiol concentrations in follicular fluid closely followed the expression patterns of steroidogenic enzymes and LH receptor mRNA. FSH receptor mRNA was present in cohorts of 2-mm follicles but declined in 4-mm follicles and was undetectable in 6- and 8-mm follicles. The expression of IGF-I and type I IGF receptor mRNA were similar for follicles of 2, 4, 6, and 8 mm. In contrast, IGF-II mRNA progressively increased in follicles collected from 2-, 4-, and 6-mm cohorts, and then decreased slightly at 8 mm. Type II IGF receptor mRNA was greatest in 8-mm follicles. IGF binding protein-2 (BP-2) mRNA decreased as follicles achieved progressively larger sizes during the preovulatory period (2 to 8 mm), whereas the IGFBP-4 mRNA remained relatively low for follicles in 2- to 6-mm cohorts but then increased markedly in 8-mm follicles. In summary, temporal and spatial patterns of gene expression for gonadotropin receptor, steroidogenic enzyme, and IGF system genes were characterized in preovulatory porcine follicles by using in situ hybridization and RNase protection analyses. The unique patterns of gene expression suggest interdependence among specific genes that may be essential for preovulatory follicular development.
卵巢卵泡生长和类固醇生成受胰岛素样生长因子(IGFs)与促性腺激素相互作用的控制。目的是确定排卵前猪卵泡发育过程中促性腺激素受体、类固醇生成酶和IGF系统基因表达的时空关系。对18头母猪进行断奶处理,并通过经直肠超声监测卵泡。当排卵前卵泡群的平均直径约为2、4、6或8mm时,从母猪采集卵巢。通过原位杂交法测定排卵前卵泡群中单个卵泡(每头母猪3至5个卵泡)的mRNA。通过对混合RNA样本的核糖核酸酶保护分析,证实了原位杂交检测到的基因表达模式。随着卵泡群平均直径从2mm增加到6mm,促黄体激素(LH)受体mRNA和类固醇生成酶mRNA(17α-羟化酶和芳香化酶)的量增加,但对于8mm的卵泡则突然下降。卵泡液中的雌二醇浓度与类固醇生成酶和LH受体mRNA的表达模式密切相关。促卵泡激素(FSH)受体mRNA存在于2mm卵泡群中,但在4mm卵泡中下降,在6mm和8mm卵泡中未检测到。2、4、6和8mm卵泡的IGF-I和I型IGF受体mRNA表达相似。相反,IGF-II mRNA在从2mm、4mm和6mm卵泡群采集的卵泡中逐渐增加,然后在8mm时略有下降。II型IGF受体mRNA在8mm卵泡中含量最高。在排卵前期(2至8mm),随着卵泡逐渐变大,IGF结合蛋白-2(BP-2)mRNA减少,而IGFBP-4 mRNA在2至6mm卵泡群中的卵泡中相对较低,但在8mm卵泡中显著增加。总之,通过原位杂交和核糖核酸酶保护分析,对排卵前猪卵泡中促性腺激素受体、类固醇生成酶和IGF系统基因表达的时空模式进行了表征。独特的基因表达模式表明特定基因之间存在相互依赖性,这可能对排卵前卵泡发育至关重要。