MONOCYTE CHEMOATTRACTANT PROTEIN 1 IN THE CORPUS LUTEUM

黄体中的单核细胞趋化蛋白 1

• 批准号: 2673908
• 负责人: PAUL LANDIS KEYES
• 金额: $ 19.45万
• 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/2673908
• 关键词: 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; monocyte chemoattractant protein 1; 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 一个短暂的腺体：在没有怀孕的情况下，黄体 在每次月经结束时从卵巢表面消失，或 发情周期这种机制是负责自发 黄体的退化在很大程度上是未知的。在一些 已经观察到种巨噬细胞在退化中积累， 黄体和其他免疫细胞的存在表明， 免疫系统在机体的破坏中起着重要作用 黄体生物信号导致了大量的 单核细胞/巨噬细胞募集到黄体结束时， 周期，以及防止他们被招募的机制 怀孕是未知的。根据该实验室最近的研究， 这个建议，我们将调查的假设，积累的 单核细胞/巨噬细胞在大鼠黄体中的作用是由 趋化因子，单核细胞趋化/活化蛋白1（MCP-1）， 这种蛋白质的表达对于生理上的 破坏黄体。 使用免疫组织化学，北方 印迹分析，原位杂交，MCP-1趋化性测定，和 黄体提取物中MCP-1的定量测定， MCP-I表达与外观之间的时间关系 单核细胞/巨噬细胞在妊娠黄体， 假孕和发情周期将被确定。的 将通过干预研究MCP-1的生理作用 （注射）与抗大鼠MCP-1抗体，以确定是否 单核细胞/巨噬细胞的募集可以被阻断， 施用抗体可防止正常的退行性变化， 组织蛋白、类固醇生成能力、芳香酶活性的丧失，以及 3B-羟类固醇脱氢酶作为黄体退化的反应 一种特殊的生理信号催乳素一个新的解释是， 提出了催乳素的矛盾促黄体/溶黄体作用 这是基于大鼠黄体退化的假设， 通过催乳素的作用介导，以刺激 MCP-I-导致单核细胞/巨噬细胞的募集/活化， 然后破坏黄体根据初步研究，我们现在知道， 催乳素刺激MCP-1的表达， 巨噬细胞在大鼠黄体。使用骑自行车的老鼠， 黄体的退化被麦角隐亭阻止， 切除垂体的大鼠，其中诱导第一次排卵并成年 垂体切除大鼠，其中黄体持续存在， 将研究催乳素给药对 诱导MCP-1的表达， 单核细胞/巨噬细胞，并诱导退行性变化， 黄体这些实验将检验一个新的假设， MCP-1在大鼠黄体中的表达及随后的黄体形成 单核细胞/巨噬细胞的募集介导生理性破坏 在大鼠黄体中，MCP-1的表达是 由催乳素调节。