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FLOW CYTOMETRY AND MOLECULAR GENETICS OF FETAL CELLS

胎儿细胞的流式细胞术和分子遗传学

基本信息

批准号：
3086862
负责人：
DIANA W. BIANCHI
金额：
$ 7.36万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-07-01 至 1990-06-30
项目状态：
已结题

项目摘要

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?

长期目标是确定激素和其他因素调节卵泡的生长和退化到各种哺乳动物的胃窦状态。的具体目标包括以下步骤： 1. 建立了大量分离纯化的方法来自猪卵巢的原始卵泡（PFS），可用于描述性和实验性研究的未探索人群分析。建议改进方法， PFS产量;对原位和游离PFS进行形态学研究 (SEM、透射电镜、厚切片（1 μ m）和H&E连续切片、组织化学和免疫细胞化学研究）;比较体外蛋白质用成熟猪卵母细胞的卵母细胞合成PF复合物，中间阶段;在存在和缺乏或适当的激素和生长因子，并分析蛋白质合成，钙调蛋白;磷积累和形态（转化鳞状前颗粒细胞变成立方形的“生长”颗粒细胞隔间？）。 2. 涉及酶促分离完整腔前卵泡的项目将继续分析短期和长期小卵泡生长对促性腺激素和生长的依赖性因素（S）支持。通过适当的激素治疗，腔前卵泡转化为功能正常的成熟腔毛囊？在短期孵育中，FSH和LH都刺激DNA 大的腔前卵泡和腔卵泡的合成，但当一起给予时， DNA合成降低到基线水平以下。有哪些机制参与FSH LH协同和拮抗作用？是肌醇脂类作为FSH/LH活动的中介，沿着营地参与其中？ 3. 在仓鼠中，几条体内和体外证据表明腔前卵泡对促性腺激素有反应，挑战了普遍认为它们的发育是非激素性的。是仓鼠是一个例外，或者是猪、大鼠、小鼠腔前卵泡 FSH和/或LH对家兔的影响？ 4. 免疫系统是以抗体的形式依赖细胞介导的毒性涉及体外卵泡闭锁和体内自发或诱发退化？巨噬细胞是关键的中介细胞吗？