HAMSTER: A MODEL FOR PROGESTERONE-DEPENDENT IMPLANTATION

描述(申请人提供)：细胞因子和生长因子在 胚胎-子宫信号在着床开始时已经被推导出来。 来自对小鼠和大鼠的研究，其中卵巢雌激素需要 开始植入。然而，目前尚不清楚相同的信号是否 该系统适用于植入可能发生在 在没有雌激素的情况下，孕酮(P4)启动子宫。申请者将会 用仓鼠作为动物模型来解决这个问题，因为像兔子一样， 豚鼠，猴子，也许还有人类，仓鼠只需要P4，但不需要 雌激素，用于建立子宫容受性和着床。然而， 仓鼠有一些共同的植入特征，比如增加了 毛细血管通透性和间质蜕膜化 植入。作为起点，申请者将专注于两个分子， 白血病抑制因子和肝素结合表皮生长因子样因子 它们对小鼠的植入很重要。申请者的初步申请 结果提示，LIF、HB-EGF及其受体在卵巢癌细胞中均有表达。 怀孕的仓鼠子宫。因此，要在此测试的工作假说 认为子宫表达的LIF和HB-EGF与其相互作用 补体受体在胚胎-子宫相互作用中的作用 仓鼠着床过程中的旁/旁分泌方式，以及这些 相互作用仅在P4的影响下发生。申请者的详细信息 1)LIF、HB-EGF在仓鼠体内的时空表达 及其受体，以及配体诱导的这些受体的磷酸化 第1-4天子宫中的受体(Erb1和ERBB4、LIF-R和GP 130) (植入前和植入后)和5-6天(植入后) 对照组和P4处理组去垂体妊娠仓鼠；2)分离 雌二醇-17β(E_2)和/或P4对子宫的交互作用 LIF、HB-EGF及其受体在去垂体仓鼠中的表达 和评价正常植入的“接受窗”以及 子宫容受性和非容受性的激素需求；以及3) LIF、HB-EGF及其受体在植入前胚胎中的表达 并用LIF和HB-EGF评价胚胎中的配体-受体信号转导 植入过程中子宫的相互作用。为了实现这些目标， 申请者将使用RT-PCR，Northern和原位杂交， 免疫细胞化学，交联，受体磷酸化，胚胎培养 和转学。预计随着更好地了解 子宫容受性发育机制与胚胎-子宫 女性植入、生育和不孕的相互作用可能是 更有效的管理。

DESCRIPTION (provided by applicant): Roles of cytokines and growth factors in embryo-uterine signaling during the onset of implantation have been derived from studies in the mouse and rat, in which ovarian estrogen is required to initiate implantation. However, it is unknown whether the same signaling systems are operative in species in which implantation can occur in the progesterone (P4) primed uterus in the absence of estrogen. The applicant will address this question using hamsters as an animal model because like rabbits, guinea pigs, monkeys and perhaps humans, hamsters require only P4, but not estrogen, for establishment of uterine receptivity and implantation. However, hamsters share some common features of implantation, such as increased capillary permeability and stromal decidualization at the site of implantation. As a starting point, the applicant will focus on two molecules, leukemia inhibitory factor (LIF) and heparin binding-EGF-like factor (HB-EGF) that are important for implantation in mice. The applicant's preliminary results suggest that LIF, HB-EGF, and their receptors are expressed in the pregnant hamster uterus. Thus, the working hypothesis to be tested in this proposal is that uterine expressed LIF and HB-EGF interact with their complementary receptors in executing embryo-uterine interactions in paracrine/juxtracrine manners during implantation in hamsters, and these interactions occur under the influence of P4 alone. The applicant's specific aims are to study in hamsters: 1) the spatiotemporal expression of LIF, HB-EGF and their receptors, as well as ligand-induced phosphorylation of these receptors (erbBl and erbB4, LIF-R and gp 130) in the uterus on days 1-4 (preimplantation and implantation) and days 5-6 (postimplantation) in normal control and in P4-treated hypophysectomized pregnant hamsters; 2) the separate and interactive effects of estradiol-17beta (E2) and/or P4 on uterine expression of LIF, HB-EGF and their receptors in hypophysectomized hamsters, and evaluation of normal "window of receptivity" for implantation as well as hormonal requirements for uterine receptivity and non-receptivity; and 3) the expression of LIF, HB-EGF and their receptors in the preimplantation embryo and evaluation of ligand-receptor signaling with LIF and HB-EGF in embryo- uterine interactions during implantation. To accomplish these aims, the applicant will use RT-PCR, Northern and in situ hybridization, immunocytochemistry, cross-linking, receptor phosphorylation, embryo culture and transfer studies. It is anticipated that with better understanding of the mechanisms of the development of uterine receptivity and embryo-uterine interactions for implantation, fertility and infertility in women could be more effectively managed.