Méndez-Herrera M C, Tamez L, Cándido A, Reyes-Esparza J A, Pedernera E
Departamento de Embriología, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico DF, 04510.
Gen Comp Endocrinol. 1998 Aug;111(2):207-15. doi: 10.1006/gcen.1998.7108.
The aim of the present study was to evaluate the effect of human follicle stimulating hormone (hFSH) on cellular proliferation in the chick embryo ovary. Chick embryos (Babcock B300) were injected on chorioallantoic membrane with a single dose of hFSH (2.0 IU/ embryo) at Days 7, 9, or 13 of incubation or with hCG (2.0 IU/embryo) at Day 13 of incubation. At 17 days of incubation and within 24 h after hatching, left ovaries were dissected and completely dissociated. Cells from the whole ovary were classified into germ cells (primary oocytes), typical steroidogenic cells, and poorly differentiated somatic cells and counted with the aid of a hemocytometer. Aliquots of the cell suspension from the whole left ovary were analyzed by flow cytometry, in order to determine the percentage of cells at each phase of the cell cycle. In addition, samples of the suspension (1.0 x 10(6 )cells) were incubated for 2 h in basal and stimulated conditions measuring 17beta-estradiol secretion in the medium. The ovarian cell number at 17 days of incubation showed that hFSH treatment at Day 7 did not modify the cell number in any of the subpopulations evaluated; treatment at Day 9 resulted in an increase in poorly differentiated somatic cell number, without changes in steroidogenic and germ cells, whereas hFSH treatment at Day 13 augmented the number of poorly differentiated, steroidogenic, and germ cells. The percentage of cells in S-phase was increased 12 and 15 h after hFSH treatment (Day 13). Secretion of 17beta-estradiol was increased in the hFSH-treated group (Day 13) measured at 17 days of incubation. The increase in cell number of the three subpopulations was still observed in the left ovary of the newly hatched chicken. Treatment with hCG at Day 13 of incubation did not change the number of poorly differentiated, steroidogenic, and germ cells in the left ovary, neither in the 17-day-old chick embryo nor in the newly hatched chicken. The 17beta-estradiol secretion in hCG-treated embryos was similar to controls. The present study is the first evidence of an effect of FSH on somatic and germ cell number, together with an increase in 17beta-estradiol production during chick embryo ovary development.
本研究的目的是评估人卵泡刺激素（hFSH）对鸡胚卵巢细胞增殖的影响。在孵化第7天、第9天或第13天，给鸡胚（巴布考克B300）的尿囊膜注射单剂量的hFSH（2.0国际单位/胚胎），或在孵化第13天注射hCG（2.0国际单位/胚胎）。在孵化第17天以及孵化后24小时内，解剖左侧卵巢并使其完全解离。将整个卵巢的细胞分为生殖细胞（初级卵母细胞）、典型的类固醇生成细胞和分化程度低的体细胞，并借助血细胞计数器进行计数。对来自整个左侧卵巢的细胞悬液进行等分试样，通过流式细胞术分析，以确定细胞周期各阶段的细胞百分比。此外，将悬浮液样本（1.0×10⁶个细胞）在基础条件和刺激条件下孵育2小时，测定培养基中17β-雌二醇的分泌量。孵化第17天时的卵巢细胞数量显示，第7天用hFSH处理并未改变所评估的任何亚群中的细胞数量；第9天处理导致分化程度低的体细胞数量增加，类固醇生成细胞和生殖细胞数量无变化，而第13天用hFSH处理增加了分化程度低的体细胞、类固醇生成细胞和生殖细胞的数量。hFSH处理后（第13天）12小时和15小时，处于S期的细胞百分比增加。在孵化第17天时测定，hFSH处理组（第13天）的17β-雌二醇分泌增加。在新孵出雏鸡的左侧卵巢中仍观察到三个亚群的细胞数量增加。在孵化第13天用hCG处理，在17日龄鸡胚和新孵出雏鸡的左侧卵巢中，均未改变分化程度低的体细胞、类固醇生成细胞和生殖细胞的数量。hCG处理胚胎中的17β-雌二醇分泌与对照组相似。本研究首次证明了FSH对鸡胚卵巢发育过程中体细胞和生殖细胞数量有影响，同时17β-雌二醇产量增加。
