Equine chorionic gonadotropin induces in vitro follicular growth from the multi-layered secondary developmental stage in cats
Equine chorionic gonadotropin induces in vitro follicular growth from the multi-layered secondary developmental stage in cats
复制标题
DOI：
10.1016/j.theriogenology.2018.09.040
复制
发表时间：
2019-01-01
期刊：
影响因子：
2.8
通讯作者：
Chatdarong, K.
中科院分区：
文献类型：
--
作者：
Chansaenroj, A.;Songsasen, N.;Chatdarong, K.
马绒毛膜促性腺激素 (eCG) 通常用于诱导几种恶臭物种发情。然而，这种促性腺激素调节猫卵泡发生的机制仍不清楚。我们研究了不同发育阶段的猫卵巢卵泡对体外补充的不同心电图浓度的反应性。从22只猫的卵巢中机械分离卵泡，并根据其形态和直径分为三个发育阶段：1）两层次级卵泡（SF），100-150μm（n = 139）； 2）多层SF，150-300μm（n=154）； 3) 早期窦卵泡 (AF)，>= 300-500 pm (n = 123)。然后将卵泡封装在 0.5% (w/v) 海藻酸钠中，并在补充有 0、0.05、0.1 或 0.511.1/mL eCG 的最低必需培养基中培养 12 天。每 3 天评估一次卵泡和卵母细胞直径。第12天，使用实时PCR分析FHSR、LHCGR、GDF9、BMPI5、CYP17A1、CYP19A1和STAR的mRNA表达水平。培养12 d后，各浓度的eCG均不影响卵泡生长和两层SF mRNA表达（P > 0.05）。然而，0.05 IU/mL 的 eCG 浓度刺激多层 SF 和早期 AF 中的卵泡生长和基因表达（P < 0.05）。相应地，用0.05 IU/mL eCG处理的多层SF和早期AF中的卵母细胞直径没有变化。从mRNA表达来看，STAR的水平在早期AF中增强（P < 0.05），并且在0.05 IU/mL eCG培养的多层SF中趋于增加（P = 0.08），而其他基因的表达不受影响。总之，从多层 SF 阶段开始，猫卵泡对心电图的反应就很明显。 0.05 IU/mL 的 eCG 补充剂似乎最适合家猫的卵泡培养。 (C) 2018 Elsevier Inc. 保留所有权利。
Equine chorionic gonadotropin (eCG) has been commonly used to induce estrus in several fetid species. However, the mechanisms by which this gonadotropin regulates cat folliculogenesis are still unclear. We investigated the responsiveness of cat ovarian follicles at different developmental stages to various eCG concentrations supplemented in vitro. Follicles were mechanically isolated from the ovaries of 22 cats and categorized into three developmental stages based on their morphology and.diameter: 1) two layered secondary follicle (SF), 100-150 mu m (n = 139); 2) multi-layered SF, 150-300 mu m (n = 154); and 3) early antral follicle (AF), >= 300-500 pm (n = 123). The follicles were then encapsulated in 0.5% (w/v) sodium alginate and cultured for 12 days in Minimum Essential Medium supplemented with 0, 0.05, 0.1 or 0.511.1/mL eCG. Follicle and oocyte diameters were assessed every 3 days. On Day 12, mRNA expression levels of FHSR, LHCGR, GDF9, BMPI5, CYP17A1, CYP19A1 and STAR were analyzed using real-time PCR. After being cultured for 12 days, follicle growth and mRNA expression of two-layered SF were not influenced by eCG at all concentrations (P > 0.05). However, the concentration of eCG at 0.05 IU/mL stimulated follicular growth and gene expressions in the multi-layered SF and early AF (P < 0.05). Correspondingly, the diameter of oocytes in the multi-layered SF and early AF treated with 0.05 IU/mL eCG was unchanged. Considering the mRNA expression, the level of STAR was enhanced in the early AF (P < 0.05) and tended to increase in the multi-layered SF (P = 0.08) cultured in 0.05 IU/mL eCG, whereas the expression of other genes was not affected. In sum, the responsiveness of cat follicles to eCG is apparent from the multi layered SF stage onward. The eCG supplementation at 0.05 IU/mL appeared to be optimal for the follicle culture in the domestic cats. (C) 2018 Elsevier Inc. All rights reserved.