HAMSTER: A MODEL FOR PROGESTERONE-DEPENDENT IMPLANTATION

描述（由申请人提供）：细胞因子和生长因子在 胚胎-子宫信号在着床开始期间已经被导出 在小鼠和大鼠的研究中，卵巢雌激素需要 开始植入然而，不知道是否相同的信令 系统在植入可以发生在 孕酮（P4）在没有雌激素的情况下启动子宫。申请人将 用仓鼠作为动物模型来解决这个问题，因为和兔子一样， 豚鼠，猴子，也许还有人类，仓鼠只需要P4， 雌激素，用于建立子宫容受性和着床。然而，在这方面， 仓鼠有一些共同的植入特征，如增加 毛细血管通透性和基质蜕膜化的网站 置入作为起点，申请人将重点关注两种分子， 白血病抑制因子（LIF）和肝素结合EGF样因子（HB-EGF） 对移植到小鼠体内很重要。 申请人的初步 结果表明，LIF，HB-EGF及其受体在细胞中表达。 怀孕仓鼠子宫因此，本研究中待检验的工作假设 提示子宫表达的LIF和HB-EGF与它们的 互补受体在执行胚胎-子宫相互作用， 在仓鼠植入过程中的旁分泌/后分泌方式，这些 相互作用仅在P4的影响下发生。申请人的具体 目的是在仓鼠中研究：1）LIF、HB-EGF的时空表达 及其受体，以及配体诱导的这些 受体（erbB 1和erbB 4，LIF-R和gp 130） （植入前和植入）和第5-6天（植入后） 对照组和P4处理的垂体切除妊娠仓鼠; 2）单独的 雌二醇-17 β（E2）和/或P4对子宫内膜的相互作用 LIF、HB-EGF及其受体在去垂体仓鼠中的表达， 和评价植入的正常“接受窗口”，以及 子宫容受性和非容受性的激素需求;和3） LIF、HB-EGF及其受体在植入前胚胎中的表达 和评估胚胎中LIF和HB-EGF的配体-受体信号传导- 着床期间子宫的相互作用。为了实现这些目标， 申请人将使用RT-PCR，北方和原位杂交， 免疫细胞化学，交联，受体磷酸化，胚胎培养 和转移研究。预计，随着对《公约》的更好理解， 子宫容受性和胚胎-子宫发育的机制 相互作用的植入，生育力和不孕症的妇女可能是 更有效地管理。

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.