Xie Meina, Zhang Caiqiao, Zeng Weidong, Mi Yuling
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, P.R. China.
Comp Biochem Physiol A Mol Integr Physiol. 2004 Dec;139(4):521-6. doi: 10.1016/j.cbpb.2004.10.017.
The effects of follicle-stimulating hormone (FSH) and 17beta-estradiol (E2) on chicken ovarian germ cell proliferation were evaluated through a germ-somatic cell coculture model. Ovarian cells from the left ovaries of 18-day-old chicken embryos were cultured in serum-free McCoy's 5A medium at 39 degrees C and challenged with FSH (0.25-1.0 IU/mL) or E2 (10(-8)-10(-5) M) alone and in combination for 48 h. The number of germ cells was counted, and the proliferating cells were immunolocalized by a specific antibody against proliferating cell nuclear antigen (PCNA). The labeling index (LI) was determined for germ cells. Results revealed that germ cells could survive and kept proliferating under support of somatic cells. Germ cells were localized by expression of a specific antibody for stem cell factor receptor c-kit. Both FSH (0.25-1.0 IU/mL) and E2 (10(-7)-10(-5) M) alone induced a marked increase in germ cell number (P<0.05), and PCNA-LI of germ cells was greater in FSH-treated groups (0.25-1.0 IU/mL) and E2-treated groups (10(-8)-10(-5) M), compared with vehicle-treated group (P<0.05). Furthermore, FSH manifested a synergistic effect with E2 (10(-6)-10(-5) M) in stimulating germ cell proliferation. These results indicate that FSH might interact with estrogen to promote ovarian germ cell proliferation in embryonic chickens near hatching.
通过生殖细胞 - 体细胞共培养模型评估促卵泡激素(FSH)和17β-雌二醇(E2)对鸡卵巢生殖细胞增殖的影响。将18日龄鸡胚左侧卵巢的细胞在无血清的McCoy's 5A培养基中于39℃培养,分别单独用FSH(0.25 - 1.0 IU/mL)或E2(10^(-8) - 10^(-5) M)以及联合使用进行处理48小时。对生殖细胞进行计数,并用抗增殖细胞核抗原(PCNA)的特异性抗体对增殖细胞进行免疫定位。测定生殖细胞的标记指数(LI)。结果显示,生殖细胞在体细胞的支持下能够存活并持续增殖。通过干细胞因子受体c-kit的特异性抗体表达对生殖细胞进行定位。单独使用FSH(0.25 - 1.0 IU/mL)和E2(10^(-7) - 10^(-5) M)均可显著增加生殖细胞数量(P<0.05),与溶剂处理组相比,FSH处理组(0.25 - 1.0 IU/mL)和E2处理组(10^(-8) - 10^(-5) M)中生殖细胞的PCNA-LI更高(P<0.05)。此外,FSH与E2(10^(-6) - 10^(-5) M)在刺激生殖细胞增殖方面表现出协同作用。这些结果表明,FSH可能与雌激素相互作用,促进接近孵化的胚胎鸡卵巢生殖细胞增殖。