Hild-Petito S, Ottobre A C, Hoyer P B
Department of Physiology, University of Arizona, Tucson 85724.
J Reprod Fertil. 1987 Jul;80(2):537-44. doi: 10.1530/jrf.0.0800537.
Peripheral blood samples were collected daily (Days 1-10 after ovulation) and analysed for progesterone content. Luteal tissue was collected on Day 10 after the LH surge, or Day 10 after hCG injection from cyclic and superovulated ewes, respectively. The tissue was enzymically dispersed and an aliquant was utilized for measurement of cell diameters, and staining for 3 beta-hydroxy-delta 5-steroid dehydrogenase-delta 5, delta 4-isomerase activity (3 beta-HSD). The remaining cell preparation was separated into small (10-22 micron) and large (greater than 22 micron) cell fractions by elutriation. Small and large cell suspensions were incubated (37 degrees C, 2 h) in the presence or absence or ovine LH (100 ng/ml) or dbcAMP (2 mM) and progesterone content of the medium was measured. Superovulation did not affect circulating progesterone concentrations, when expressed per mg luteal tissue recorded; basal progesterone production by small or large luteal cells; the unresponsiveness of large luteal cells to ovine LH or dbcAMP; the ratio of small:large cells recovered by dissociation the mean diameter of total cells; or the mean diameter of large cells. However, the mean cell diameter and LH stimulation of progesterone production by small cells were greater (P less than 0.05) in luteal tissue collected from superovulated than in that from cyclic ewes. These differences appear to be an amplification of basic function. Therefore, we conclude that corpora lutea obtained from superovulated ewes can be used to study functional aspects of small and large cells.
每天采集外周血样本（排卵后第1天至第10天）并分析孕酮含量。分别从周期性发情和超数排卵的母羊中，在促黄体生成素（LH）高峰后第10天或人绒毛膜促性腺激素（hCG）注射后第10天采集黄体组织。将组织进行酶解分散，取一份用于测量细胞直径，并对3β-羟基-δ5-类固醇脱氢酶-δ5,δ4-异构酶活性（3β-HSD）进行染色。剩余的细胞制剂通过淘析法分离成小细胞（10 - 22微米）和大细胞（大于22微米）部分。将小细胞和大细胞悬液在有或无羊LH（100纳克/毫升）或二丁酰环磷腺苷（dbcAMP，2毫摩尔）存在的情况下孵育（37摄氏度，2小时），并测量培养基中的孕酮含量。当以每毫克记录的黄体组织表示时，超数排卵不影响循环孕酮浓度；小或大黄体细胞的基础孕酮产生；大黄体细胞对羊LH或dbcAMP无反应性；通过解离回收的小细胞与大细胞的比例、总细胞的平均直径或大细胞的平均直径。然而，从超数排卵母羊采集的黄体组织中，小细胞的平均细胞直径和LH对孕酮产生的刺激作用比从周期性发情母羊采集的黄体组织更大（P < 0.05）。这些差异似乎是基本功能的放大。因此，我们得出结论，从超数排卵母羊获得的黄体可用于研究小细胞和大细胞的功能方面。
