FLOW CYTOMETRY AND MOLECULAR GENETICS OF FETAL CELLS

长期目标是确定荷尔蒙和其他因素 从原始开始调控卵泡的生长和退化 在不同种类的哺乳动物中处于有腔状态。具体目标 包括： 1.提出了一种大规模分离纯化的方法 猪卵巢中的原始卵泡(PFS)到目前为止是这样的 未开发的人群可用于描述性和试验性 分析。建议改进方法，以增加 PFS的产率；对原位和游离的PFS进行形态研究 (扫描电子显微镜、透射电子显微镜、厚切片(1微米)和HE连续切片、组织化学 和免疫细胞化学研究)；比较体外蛋白质 猪成熟卵母细胞与卵母细胞的PF复合体的合成 中期；在有条件的情况下培养PFS数天 缺乏或适当的激素和生长因子，并分析变化 蛋白质合成，钙调素；磷的积累和形态(转化 鳞状颗粒前细胞形成立方“生长”的颗粒 车厢？)。 2.涉及用酶法分离的完整腔前卵泡的项目 从仓鼠卵巢中继续分析短期和长期 小卵泡对促性腺激素和生长的依赖性 因素(S)支持。在适当的激素治疗下可以培养出 腔前卵泡转化为功能正常的成熟腔 毛囊？在短期孵育中，促卵泡激素和促黄体生成素都能刺激DNA 腔前和腔前大卵泡的合成，但同时给予时 DNA合成降低到基线水平以下。其机制是什么？ 参与促黄体生成素-黄体生成素的协同和拮抗？是肌醇类脂类 与cAMP一起参与FSH/LH活性的中介吗？ 3.在仓鼠体内，体内和体外的几种证据 表明腔前卵泡对促性腺激素有反应 挑战了普遍认为他们的发育是非荷尔蒙的。是 仓鼠是猪、大鼠、小鼠腔前卵泡的例外 和/或促卵泡激素和/或促黄体生成素对兔的影响？ 4.免疫系统是以抗体依赖细胞的形式介导的吗 毒性涉及体外卵泡闭锁和体内自发性卵巢癌 诱发退行性变？巨噬细胞是关键的中介细胞吗？

The long term objective is to determine the hormonal and other factors regulating the growth and regression of ovarian follicles from primordial to antral states in various species of mammals. The specific objectives include: 1. A method has been worked out to isolate and purify in large numbers primordial follicles (PFS) from pig ovaries which makes this hitherto unexplored population available for descriptive and experimental analyses. It is proposed to improve the methodology to increase the yield of PFS; carry out morphological studies on in situ and freed PFS (SEM, TEM, thick sections (1 mu m) and H&E serial sections, histochemical and immunocytochemical investigations); compare in vitro protein synthesis of the PF complex with oocytes of mature pig oocytes and intermediate stages; to culture PFS for several days in the presence and absence or appropriate hormones and growth factors and analyze changes in protein synthesis, cam;p accumulation and morphology (transformation of squamous pregranulosa cells into a cuboidal "growing" granulosal compartment?). 2. Projects involving enzymatically isolated intact preantral follicles from hamster ovaries will be continued to analyze short and long term dependency of small growing follicles on gonadotropin and growth factor(s) support. With suitable hormonal treatments can cultured preantral follicles be converted to functionally normal mature antral follicles? In short term incubations, FSH and LH both stimulate DNA synthesis of large preantral and antral follicles but when given together DNA synthesis is reduced below baseline levels. What are the mechanisms involved in FSH-LH synergism and antagonism? Are inositol lipids involved, along with camp, as intermediaries of FSH/LH activity? 3. In the hamster, several lines of in vivo and in vitro evidence indicate that preantral follicles are responsive to gonadotropins which challenges the prevalent belief that their development is anhormonal. Is the hamster an exception or are preantral follicles in pig, rate, mouse and rabbit influenced by FSH and/or LH? 4. Is the immune system in the form of antibody - dependent cell mediated toxicity involved in vitro follicular atresia and in vivo spontaneous or induced degeneration? Are macrophages the key intermediary cell?