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西部斑臭鼬植入前子宫和囊胚中表皮生长因子受体的表达

Expression of epidermal growth factor receptor in the preimplantation uterus and blastocyst of the western spotted skunk.

作者信息

Paria B C, Das S K, Mead R A, Dey S K

机构信息

Department of Obstetrics & Gynecology, University of Kansas Medical Center, Kansas City.

出版信息

Biol Reprod. 1994 Aug;51(2):205-13. doi: 10.1095/biolreprod51.2.205.

Abstract

The western spotted skunk is unique in that its blastocysts undergo a 180-220-day period of arrested development before implantation. We investigated the potential role of epidermal growth factor (EGF)-related growth factors in regulating uterine and embryonic development in this species by studying the status of EGF receptor (EGF-R) in these tissues during delayed implantation and resumption of embryonic development. The cell-specific distribution of EGF binding sites and the expression of EGF-R mRNA were assayed by autoradiography and Northern blot analysis, respectively. The size of EGF-R was determined by affinity cross-linking studies, and its bioactivity was examined by determining EGF-dependent subcellular protein tyrosine kinase (PTK) activity. EGF binding sites were localized in the uterine luminal and glandular epithelium, endometrial stroma, myometrium, and blood vessels during both stages of pregnancy. As examined by Northern blot hybridization, a cRNA probe specific to mouse EGF-R hybridized to poly(A)+ RNA of skunk uteri. Transcripts similar to those of mouse uterine EGF-R were identified. [125I]-EGF was cross-linked to a 170-kDa protein both in the uterus and in blastocysts collected during the delayed implantation and periimplantation periods. However, EGF-induced PTK activity was significantly elevated above background levels during the period of renewed embryonic development, but not during arrested embryonic development. The results suggest that EGF-related growth factors may play an important role in regulating embryonic development in this species and that a change in the number and/or functional status of the EGF-R may be a prerequisite for blastocyst activation and implantation in the spotted skunk.

摘要

西部斑点臭鼬的独特之处在于其胚泡在植入前会经历180 - 220天的发育停滞期。我们通过研究延迟植入和胚胎发育恢复期间这些组织中表皮生长因子(EGF)受体(EGF-R)的状态,来探究EGF相关生长因子在调节该物种子宫和胚胎发育中的潜在作用。分别通过放射自显影和Northern印迹分析来检测EGF结合位点的细胞特异性分布和EGF-R mRNA的表达。通过亲和交联研究确定EGF-R的大小,并通过测定EGF依赖性亚细胞蛋白酪氨酸激酶(PTK)活性来检测其生物活性。在妊娠的两个阶段,EGF结合位点均定位于子宫腔上皮、腺上皮、子宫内膜基质、子宫肌层和血管中。通过Northern印迹杂交检测,小鼠EGF-R特异性的cRNA探针与臭鼬子宫的poly(A)+ RNA杂交。鉴定出了与小鼠子宫EGF-R相似的转录本。在延迟植入期和植入前期收集的子宫和胚泡中,[125I]-EGF均与一种170 kDa的蛋白质交联。然而,在胚胎发育重新开始期间,EGF诱导的PTK活性显著高于背景水平,而在胚胎发育停滞期间则不然。结果表明,EGF相关生长因子可能在调节该物种胚胎发育中起重要作用,并且EGF-R数量和/或功能状态的变化可能是斑点臭鼬胚泡激活和植入的先决条件。

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