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Expression, distribution and function of the focal adhesion kinase (pp125^(FAK)) during murine ectoplacental cone outgrowth.in vitro

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Chinese Science Bulletin 1998年第17期43卷 1473-1480页

作者： ZHANG Chunyu DUAN Enkui CAO Yujing ZENG Guoqing State Key Laboratory of Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100080China Boyer Center for Molecular Medicine Department of GeneticsYale University School of MedicineNew HavenCT 06510USA

Mouse embryo implantation is a complex process that includes trophoblast cells derived from ectoplacental cone (EPC) adhesion to and migration through the extracellular matrix (ECM) of uterine endometrium and invasion into the decidua. At the time of implantation, fibronectin (FN) is abundant in the decidua and is distributed pericellularly around each individual stromal cell, and its receptor (integrin α-5β-1) expression on trophoblast populations is up-regulated. The focal adhesion kinase, a 125 ku protein tyrosine kinase (pp125 FAK), is tyrosine phosphorylated upon integrin engagement with its ECM ligand, and its tyrosine phosphorylation sites then serve as the binding sites which couple it with cellular proteins that contain Src SH2 or SH3 domains. Through these linkages, pp125 FAK may integrate multiple signals triggered by integrins. The model of EPC culture %in vitro% was used to study the expression, distribution and function of pp125 FAK during EPC outgrowth.on FN. Results indicated that, pp125 FAK primarily expressed and distributed in cellular focal adhesions of the front edge of trophoblast outgrowth.from EPC, and was localized in the peripheral region of the individual migrating trophblast cell; antibody or antisense oligodeoxynucleotide to pp125 FAK inhibited EPC attachment and outgrowth. as well as trophoblast cells spreading and migration. This experiment demonstrated that pp125 FAK as an integrin-mediated signaling molecule was involved in EPC outgrowth.%in vitro%, and played an important role during trophoblast cells interaction with FN.

关键词： fibronectin integrin focal adhesion kinase(pp125 FAK) ectoplacental cone attachment outgrowth.

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The expression and function of VEGF at embryo implantation "window" in the mouse

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Chinese Science Bulletin 2001年第5期46卷 409-412,441页

作者： Jing Zhang Li Wang Liquan Cai Yujing Cao Enkui Duan Chinese Acad Sci Inst Zool State Key Lab Reprod Biol Beijing 100080 Peoples R China

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen that plays a critical role in angiogenesis. Recent reports indicated that VEGF was closely involved in embryo implantation and embryonic vasculogenesis. However, very little information is available about the detailed expression and function of VEGF at implantation 'window'. In this work, VEGFs were primarily present on uterine epithelial cell monolayer and blasto-cysts including the outgrew trophoblasts at implantation window. VEGF antibodies decreased the number of mice embryos implanted and the percentage of blastocysts with attachment and outgrowth.in a co-culture model in a dose-dependant manner. These findings demonstrate that VEGF is one of the essential cytokines for embryo implantation in mouse. VEGF may act as a local mediator to regulate the maternal-fetal interaction, and facilitate blastocyst implantation.

关键词： vascular endothelial growth factor embryo implantation trophoblast attachment outgrowth.

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Expression of recombinant rat Neurotrophin-3 in Chinese hamster ovary cells

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Science China(Life Sciences) 1999年第6期42卷 655-662页

作者：季爱民舒斯云包新民邹恒琴张忠义李明

The CHO cell line stably producing recombinant rat NT-3 was established. The insertion of rNT-3 cDNA into transferred cell gonome was analyzed with Southern blot. The expressed protein was identified by Dot ELISA (enzyme-linked immunosorbent assay) and Western blot. Western blot showed a clear specific band of about 14 ku for NT-3. The mean level of rNT-3 in four NT-3cDNA/CHO cell lines was about 2 100 ng/106 cells/48 h determined by EIA. The conditioned-medium (CM) of NT-3cDNA/CHO cells could promote the fiber outgrowth.of the dissociated dorsal root ganglion of 8-day-old chick embryos, which shows a dose-response relationship. A half-maximal concentration of the biological activity (EC50) of the recombinant protein was approximately 16.7 ng/mL. The MoAb 3W3 of NT-3 could neutralize the biological activity of the rNT-3.

关键词： Neurotrophin-3 cDNA recombinant nerve fiber outgrowth.

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Effect of Gossypol on Bovine Blastocyst Attachment and Trophoblastic outgrowth.in Vitro

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Developmental and Reproductive Biology 1993年第2期2卷 43-45页

作者：曾国庆 Gurpreet Singh N.L.First State Key Laboratory of Reproductive Biology Department of Endocrinology Institute of Zoology Academia Sinica Beijing 100080 Embryo Biotechnology Laboratory National Institute of Immunology New Delhi India 110067 Department of Meat and Animal Science University of Wisconsin-Madison U. S. A. WI 53706.

The presont study was carried out to investigate the effect of different amounts of gossypol on bovineblastocyst attachment and trophoblastic outgrowth.in vitro. Bovine oocyte were collected from theovaries of slaughtered cows and were matured and fertilized in vitro. Cleaved oocyte were culturedin CRlaa + BOEC and TC-199 + 10% FCS combined in an 1:1 ratio. After 8 days of co-culture,the hatched blastocysts were randomly allotted to different treatment groups. All were cultured ona fetal fibroblast monolaycr (prepared from bovine fetuses) in TC-199 culture medium supplementedwith 10% fetal calf serum (TCFCS). But the groups differed from one another in the dose ofgossypol given: 0.01 μg, 0.1 μg, 1 μg, 10 μs/ml, and no gossypol as control. All cultures wereperformed in 24-well culture plates at 39℃ with 5% CO_2 in air. The results indicate that the ratesof attached and outgrowing blastocysts in the medium containing 1 μg/ml gossypol were significantlylowcr than the control group (poutgrowth.were inhibited by gossypol in a dose-dependent manner.

关键词： Gossypol Blastocyst Implantation Attachment outgrowth.

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