CYTOCHROME-P-450SCC, CYTOCHROME-P-45017-ALPHA, ADRENODOXIN, AND REDUCED NICOTINAMIDE ADENINE-DINUCLEOTIDE PHOSPHATE-CYTOCHROME-P-450 REDUCTASE IN BOVINE FOLLICLES AND CORPORA-LUTEA - CHANGES IN SPECIFIC CONTENTS DURING THE OVARIAN CYCLE
CYTOCHROME-P-450SCC, CYTOCHROME-P-45017-ALPHA, ADRENODOXIN, AND REDUCED NICOTINAMIDE ADENINE-DINUCLEOTIDE PHOSPHATE-CYTOCHROME-P-450 REDUCTASE IN BOVINE FOLLICLES AND CORPORA-LUTEA - CHANGES IN SPECIFIC CONTENTS DURING THE OVARIAN CYCLE
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DOI：
10.1210/endo-118-4-1366
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发表时间：
1986-04-01
期刊：
影响因子：
4.8
通讯作者：
SIMPSON, ER
中科院分区：
文献类型：
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作者：
RODGERS, RJ;WATERMAN, MR;SIMPSON, ER
为了研究牛动情周期期间卵巢类固醇产生模式的基础，估计了主要类固醇生成酶、17α-羟化酶细胞色素 P-450 和胆固醇侧链裂解细胞色素 P-450 (细胞色素 P-450scc) 及其各自电子供体、NADPH-细胞色素 P-450 还原酶和肾上腺氧还蛋白的组织浓度，并与非类固醇生成酶的组织浓度进行了比较。酶，细胞色素c氧化酶和F-ATP酶。通过离心去除卵泡液后，在中等大小 (9-11 mm) 和大 (14-18 mm) 卵泡中以及黄体期早期、早中、晚中和晚期的黄体中估计这些酶的水平（每组 n = 5）。除细胞色素c氧化酶通过测定比活性进行定量外，所有酶和电子供体的具体含量均通过免疫印迹分析测定。 17α/羟化酶细胞色素P-450的特异性（每微克组织匀浆蛋白）和总（每个卵泡或黄体）组织含量从中型卵泡到大卵泡分别增加0.5倍和5倍，但随后在黄体期早期在黄体中降低至检测不到的水平，并且在整个黄体期保持检测不到。相反，NADPH-细胞色素P-450还原酶的具体含量在卵泡和黄体之间相似。卵泡中细胞色素P-450scc、肾上腺素和细胞色素c氧化酶的具体含量与黄体早期黄体相似。然而，到黄体期早中期，黄体细胞色素 P-450scc（490 .+-. 46 与 5709 .+-. 982 cpm/.mu.g 蛋白质）和肾上腺氧还蛋白（44 .+-. 15 与 705 .+-. 229 cpm/.mu.g 蛋白质）的具体含量增加了 12-和分别为 15 倍（P < 0.05）。相反，细胞色素c氧化酶活性（29.1.+-.10.1 vs. 108.6.+-.20.7nmol/mg组织蛋白.cntdot.min）和F1-ATP酶的比含量仅增加3至4倍，反映了线粒体数量的增加。这些酶的水平一直保持升高，直到黄体期晚期，此时它们显着下降。结论是，排卵后诱导P-450scc和肾上腺素合成是特异性的，不仅仅反映黄体化过程中线粒体的生物发生。此外，在整个动情周期中卵巢室产生的类固醇类型的变化反映了类固醇生成酶的组织含量的变化。
To investigate the basis for the pattern of ovarian steroid production during the bovine estrous cycle, the tissue concentrations of major steroidogenic enzymes, 17.alpha.-hydroxylase cytochrome P-450 and cholesterol side-chain cleavage cytochrome P-450 (cytochrome P-450scc), and their respective electron donors, NADPH-cytochrome P-450 reductase and adrenodoxin, were estimated and compared with those of the nonsteroidogenic enzymes, cytochrome c oxidase and F-ATPase. The levels of these enzymes were estimated in medium sized (9-11 mm) and large (14-18 mm) folicles after removal of follicular fluid by centrifugation, and corpora lutea from the early, early-mid, late-mid, and late stages of the luteal phase (n = 5 per group). The specific contents of all enzymes and electron donors were determined by immunoblot analysis, except for cytochrome c oxidase, which was quantified by determination of specific activity. The specific (per microgram of tissue homogenate protein) and total (per follicle or corpus luteum) tissue contents of 17.alpha./hydroxylase cytochrome P-450 increased 0.5- and 5-fold respectively from medium sized to large follicles, but then decreased to undetectable levels in corpora lutea in the early luteal phase, and remained undetectable throughout the luteal phase. In contrast, the specific content of NADPH-cytochrome P-450 reductase was similar between follicles and corpora lutea. The specific contents of cytochrome P-450scc, adrenodoxin and cytochrome c oxidase in follicles were similar to those of corpora lutea of the early luteal phase. However, by the early-mid luteal phase the specific contents of luteal cytochrome P-450scc (490 .+-. 46 vs. 5709 .+-. 982 cpm/.mu.g protein) and adrenodoxin (44 .+-. 15 vs. 705 .+-. 229 cpm/.mu.g protein) were increased, by 12- and 15-fold, respectively (P < 0.05). In contrast, cytochrome c oxidase activity (29.1 .+-. 10.1 vs. 108.6 .+-. 20.7 nmol/mg tissue protein .cntdot. min) and the specific content of F1-ATPase increased only 3- to 4-fold, reflective of an increase in numbers of mitochondria. The levels of these enzymes remained elevated until the late luteal phase when they declined markedly. It is concluded that the induction of synthesis of P-450scc and adrenodoxin after ovulation is specific and does not merely reflect biogenesis of mitochondria during luteinization. Moreover the changes in the types of steroids produced by the ovarian compartments throughout the estrous cycle are a reflection of changes in the tissue content of steroidogenic enzymes.