MONOCYTE CHEMOATTRACTANT PROTEIN 1 IN THE CORPUS LUTEUM

黄体中的单核细胞趋化蛋白 1

• 批准号: 2403548
• 负责人: PAUL LANDIS KEYES
• 金额: $ 18.73万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2403548
• 关键词: chemoattractants; chemotaxis; corpus luteum; enzyme linked immunosorbent assay; gene expression; genetic regulation; hydroxysteroid dehydrogenases; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; leukocyte activation /transformation; macrophage; monoclonal antibody; monocyte; neutralizing antibody; northern blottings; prolactin; protein structure function; western blottings

One of the most remarkable features about the corpus luteum is that it is a transient gland: in the absence of conception, the corpus luteum disappears from the surface of the ovary at the end of each menstrual or estrous cycle. The mechanisms that are responsible for the spontaneous involution of the corpus luteum are largely unknown. In a number of species macrophages have been observed to accumulate in the regressing corpus luteum, and other immune cells are present suggesting that the immune system has an important role in the destruction of the corpus luteum. The biological signals that are responsible for the massive recruitment of monocytes/macrophages into the corpus luteum at the end of the cycle, and the mechanisms which prevent their recruitment during pregnancy are unknown. Based upon recent studies from this laboratory, in this proposal we will investigate the hypothesis that the accumulation of monocytes/macrophages in the rat corpus luteum is mediated by the chemokine, monocyte chemoattractant/activating protein l (MCP-l), and that the expression of this protein is essential for the physiological destruction of the corpus luteum. Using immunohistochemistry, Northern blot analysis, in situ hybridization, a MCP-l chemotaxis assay, and an assay for quantitation of MCP- l in extracts of corpora lutea, the temporal relationship between the expression of MCP-I and the appearance of monocytes/macrophages in the corpus luteum of pregnancy, pseudopregnancy, and of the estrous cycle will be determined. The physiological role of MCP-l will be investigated by intervention (injection) with anti-rat MCP-l antibody, to determine whether the recruitment of monocytes/macrophages can be blocked, and whether the administration of antibody prevents normal regressive changes such as the loss of tissue protein, steroidogenic capacity, aromatase activity, and 3B-hydroxysteroid dehydrogenase as the corpus luteum regresses in response to a specific physiological signal, prolactin. A new explanation is proposed for the paradoxical luteotrophic/luteolytic effect of prolactin which is based upon the hypothesis that luteal regression in the rat is mediated through an action of prolactin to stimulate the expression of MCP- I- leading to recruitment/activation of monocytes/macrophages, which then destroy the corpus luteum. From preliminary studies we now know that prolactin stimulates the expression of MCP-l and a massive invasion of macrophages in the rat corpus luteum. Using cycling rats in which regression of the corpus luteum is prevented by ergocryptine, immature hypophysectomized rats in which a first ovulation is induced and adult hypophysectomized rats in which the corpora lutea persist, the effects of prolactin administration will be investigated with respect to the induction of expression of MCP-I, the recruitment of monocytes/macrophages, and the induction of regressive changes in the corpora lutea. These experiments will test a new hypothesis that expression of MCP- l in the rat corpus luteum and the subsequent recruitment of monocytes/macrophages mediate the physiological destruction of the corpus luteum, and that in the rat, the expression of MCP-1 is regulated by prolactin.

黄体最显著的特征之一是它是 暂时性腺体：在没有受孕的情况下，黄体 在每次月经结束时从卵巢表面消失或 发情周期。负责自发行为的机制 黄体退化在很大程度上是未知的。在许多情况下 观察到不同种类的巨噬细胞在退化过程中积累 黄体和其他免疫细胞的存在，表明 免疫系统在人体的破坏中起着重要作用。 黄体。造成大规模爆炸的生物信号 月经结束时单核/巨噬细胞在黄体中的募集 周期，以及防止他们在 怀孕的人是未知的。根据该实验室最近的研究，在 在这个提议中，我们将调查这样一种假设，即 大鼠黄体中的单核/巨噬细胞是由 趋化因子、单核细胞趋化因子/激活蛋白L(MCP-L)，以及 该蛋白的表达对机体的生理功能是必不可少的。 黄体破坏。使用免疫组织化学，Northern 印迹分析、原位杂交、单核细胞趋化蛋白-L实验和An 黄体提取液中多氯酚-L的含量测定 单核细胞趋化蛋白-I的表达与肿瘤外观的时间关系 在妊娠黄体中的单核/巨噬细胞， 假孕和发情周期将被确定。这个 单核细胞蛋白质-L的生理作用将通过干预进行调查 (注射)用抗大鼠单核细胞趋化蛋白-L抗体，以确定是否 单核/巨噬细胞的募集能否被阻断，以及是否 注射抗体可防止正常的退行性改变，如 组织蛋白、类固醇合成能力、芳香酶活性丧失，以及 黄体退化时的3B-羟基类固醇脱氢酶反应 一种特定的生理信号，催乳素。一种新的解释是 催乳素的促黄体/促黄体作用 这是基于一种假设，即大鼠的黄体退化是 通过催乳素的作用来刺激 MCP-I-导致单核/巨噬细胞的募集/激活，从而 然后破坏黄体。根据初步研究，我们现在知道， 催乳素刺激单核细胞趋化蛋白-L的表达及卵巢癌细胞的大量侵袭 大鼠黄体内巨噬细胞。使用骑自行车的大鼠 未成熟的麦角隐亭可防止黄体退化 诱导首次排卵的垂体切除大鼠和成年大鼠 黄体持续存在的去垂体大鼠对黄体的影响 将对催乳素管理进行调查 诱导单核细胞趋化蛋白-I的表达，募集 单核/巨噬细胞，并诱导细胞的退行性变化 黄体。这些实验将检验一个新的假设 单核细胞趋化蛋白-L在大鼠黄体及随后黄体中的表达 单核/巨噬细胞募集介导生理性破坏 在大鼠黄体组织中，MCP-1的表达为 由催乳素调节的。