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PHYSIOLOGY OF PITUITARY FOLLICULOSTELLATE CELLS

垂体滤泡细胞的生理学

基本信息

批准号：
3320819
负责人：
JAMES JEFFREY MULCHAHEY
金额：
$ 6.88万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-12-01 至 1989-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3320819
关键词：
cyclic AMP electron microscopy follicle stimulating hormone hormone regulation /control mechanism laboratory rat luteinizing hormone physiology pituitary gonadal axis prolactin radiotracer somatotropin tissue /cell culture

项目摘要

The long range goal is to more fully understand the physiology of the pituitary folliculostellate (FS) cell. No unequivocal function has been demonstrated for these cells. The hypothesis to be tested is that FS cells affect anterior pituitary (AP) physiology by altering the local extra- or intracellular levels of 3', 5' cyclic adenosine monophosphate (cAMP) and Ca2+. Altered local levels of cAMP and Ca2+ then affect the rate of hormone secretion from secretory AP cells. The specific aims are to: 1) develop and validate an in vitro cell culture paradigm for rat FS cells, 2) describe the morphology and ultrastructure of FS cells in these cultures and analyze the patterns and nature of FS cell association with AP cells, 3) determine the effects of FS cells on hormone secretion from other AP cells and 4) elucidate the mechanisms by which FS cells might communicate with other FS cells and influence hormone secretion from AP cells. These aims will be achieved by developing monolayer cultures of FS cells on an extracellular matrix. Highly enriched populations of FS cells will be obtained by separation of monodispersed pituitary cells by unit gravity sedimentation. These cells will be used then to initiate the monolayer FS cell cultures. The monolayer cultures will be used for studies of cAMP production in basal and hormone-stimulated states. Intracellular Ca2+ mobilization will also be examined in these states with Quin 2 fluorescence. Cell-cell connections will be examined using a dye coupling model. Finally AP hormone (growth hormone, luteinizing hormone and prolactin) secretion from individual AP cells, either associated with FS cells or unassociated, will be examined with reverse hemolytic plaque assays for these hormones. The effects of FS cells on hormone secretion, secretagogue responsiveness and the spectrum of secretagogues to which AP cells respond will be investigated. These studies of FS cell physiology are likely to provide additional insight into the growth of the AP in fetal development and various neoplastic conditions.

长期目标是更全面地了解垂体滤泡星状（FS）细胞。没有明确的功能，为这些细胞展示。待检验的假设是FS细胞影响垂体前叶（AP）的生理学改变当地的额外或细胞内3 '，5'环磷酸腺苷（cAMP）水平和 Ca2+。局部cAMP和Ca 2+水平的改变会影响分泌型AP细胞分泌激素。具体目标是：1）开发并验证大鼠FS细胞的体外细胞培养模式，2）描述了这些培养物中FS细胞的形态和超微结构并分析FS小区与AP小区关联的模式和性质， 3)确定FS细胞对其他AP激素分泌的影响细胞和4）阐明FS细胞可能沟通的机制与其他FS细胞和影响激素分泌的AP细胞。这些目的将通过开发FS细胞的单层培养物来实现，细胞外基质高度富集的FS细胞群将被通过单位重力分离单分散的垂体细胞获得沉淀然后将这些细胞用于启动单层FS 细胞培养单层培养物将用于cAMP的研究在基础状态和刺激状态下的生产。细胞内ca 2 + 在这些州，还将用奎因2号来检查动员情况荧光。将使用染料偶联检查细胞-细胞连接模型最后是AP激素（生长激素、促黄体生成激素和催乳素）分泌的个别AP细胞，或与FS 细胞或不相关，将用反向溶血空斑进行检查检测这些激素。 FS细胞对激素分泌的影响，促分泌素反应性和AP对促分泌素的谱将研究细胞反应。这些FS细胞生理学的研究有可能为胎儿AP的生长提供额外的见解，发展和各种肿瘤条件。