Avila Rodolfo, Samar María E, Ferraris Ricardo, Centurión Claudio
Rev Fac Cien Med Univ Nac Cordoba. 2002;59(1):63-9.
Several authors have shown the effect of LH or hCG gonadotrophins on steroid secretion in the chick embryo ovary. However, the effects of luteinising hormone or human chorionic gonadotropin on chick embryo ovary innervation have not been studied. The purpose of the present work was to study the subcellular aspects of interstitial cells innervation in chick embryo ovaries cultured with LH or hCG. Coob's White Rock female chick embryos at 7, 11, 15 and 19 days of incubation (in ovo) were employed. Explants of the left and right ovaries were cultured separately for 4 days in: 1--Basic medium (control): minimum essential medium (MEM-GIBCO) with 10% bovine fetal serum, 2--Basic medium with added LH or hCG (problems). The cultures were processed to be studied ultrastructurally (TEM). Left and right ovary explants were fixed two hours at room temperature in Karnovsky's fixative. The tissues were then post-fixed in osmium tetroxide in 0.1 M cacodylate buffer for one hour and after a rinse in buffer were dehydrated in acetone: afterwards they were embedded in Araldite resin. Thick sections stained with 1% Toluidine blue were examined and photographed using a Zeiss Photo II microscope. Thin sections were stained with uranyl acetate and lead citrate and examined in a Siemens ElmisKop 101 electron microscope.
In both ovaries the isolated interstitial cells and the appearance of nerve fibers and nerve endings were observed in the explants of 11 day cultured during four days and in a similar way at the age of 15 days of in ovo development.
With LH or hCG nerve fibers and nerve endings were observed both in the right gonad and in the medulla of the left ovary in close contact with the steroid-producing interstitial cells. Ultrastructurally, the interstitial cells are recognized by their typical organoids: SER, Golgi apparatus, mitochondria with tubular crests and abundant lipidic droplets. The innervation of these cells were seen in ovaries at 7 days development, cultured during 4 days. The present state corresponds to that of 11 days development in ovo. The nerve endings had mitochondria, microtubules and small and medium-sized vesicles, some with an electrodense content. These results in vitro suggest that innervation of the ovaries is controlled by an indirect mechanism via the hypothalamic-pituitary system and another local with production neurotrophic factors modulated by LH.
几位作者已经展示了促黄体生成素（LH）或人绒毛膜促性腺激素（hCG）对鸡胚卵巢类固醇分泌的影响。然而，促黄体生成素或人绒毛膜促性腺激素对鸡胚卵巢神经支配的影响尚未得到研究。本研究的目的是研究用LH或hCG培养的鸡胚卵巢间质细胞神经支配的亚细胞方面。使用孵化7、11、15和19天（在卵内）的科布氏白洛克雌性鸡胚。左右卵巢外植体分别在以下培养基中培养4天：1 - 基础培养基（对照）：含10%胎牛血清的最低必需培养基（MEM - GIBCO），2 - 添加LH或hCG的基础培养基（实验处理）。培养物经过处理以进行超微结构研究（透射电子显微镜）。左右卵巢外植体在室温下用卡诺夫斯基固定液固定两小时。然后组织在0.1M二甲胂酸盐缓冲液中的四氧化锇中后固定一小时，在缓冲液中冲洗后在丙酮中脱水：之后它们被包埋在阿拉代特树脂中。用1%甲苯胺蓝染色的厚切片用蔡司Photo II显微镜检查并拍照。薄切片用醋酸铀和柠檬酸铅染色，并在西门子ElmisKop 101电子显微镜下检查。
在两个卵巢中，在培养4天的11日龄外植体以及卵内发育15日龄时以类似方式观察到分离的间质细胞以及神经纤维和神经末梢的出现。
在右侧性腺和左侧卵巢髓质中均观察到与产生类固醇的间质细胞紧密接触的LH或hCG神经纤维和神经末梢。在超微结构上，间质细胞通过其典型的细胞器来识别：滑面内质网、高尔基体、具有管状嵴的线粒体和丰富的脂滴。在发育7天、培养4天的卵巢中可见这些细胞的神经支配。目前的状态相当于卵内发育11天的状态。神经末梢有线粒体、微管以及中小泡，一些小泡含有电子致密内容物。这些体外实验结果表明，卵巢的神经支配由下丘脑 - 垂体系统通过间接机制控制，以及由LH调节产生神经营养因子的另一种局部机制控制。
