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Somatic cell bovine cloning:Effect of donor cell and recipients

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Chinese Science Bulletin 2003年第6期48卷 549-554页

作者： CHENDayuan LIJinsong 等 statekeylaboratoryofreproductivebiology InstiuteofZoologyChineseAcademyofSciencesBeijing100080China statekeylaboratoryofreproductivebiology InstiuteofZoologyChineseAcademyofSciencesBeijing100080

Adult somatic cell nuclear transfer was con-ducted by using cultured ear fibroblast cells obtained from a Holstein female cow (GN) and a Galoway herd bull (GLV). The percentages of reconstructed eggs developed into blas-tocysts were similar in GN (23.98%, 123 of 513) and in GLV groups (29.55%, 138 of 467). However, the rate of recon-structed female (GN) embryos developed into term was higher than that of male (GLV) (8.02% and 1.82%, respec-tively). Three kinds of cows, Luxi Yellow cows, Holstein heifers and Holstein cows with normal reproductive records were used as recipients. When the reconstructed embryos from GN were transferred, there was no difference in the pregnancy rate among three kinds of recipients, but the abortion rate of Luxi Yellow cows was significantly higher (85.71%) than in the other two groups (14.29% and 0%, respectively; P < 0.05). And the percentages of newborn calves in transferred embryos were significantly different between Luxi Yellow cows and Holstein breed (1.54%, 10.39% and 20.0%, respectively, P < 0.05). However, when reconstructed embryos from GLV were transferred, there was no difference among three kinds of recipients in the pregnancy rate, the abortion rate and the delivery rate.

关键词：成年牛体细胞克隆核移植供体细胞受体

Functions of VEGF in female reproductive system

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Chinese Science Bulletin 2003年第3期48卷 217-222页

作者： QIANDong ZHUCheng statekeylaboratoryofreproductivebiology InstituteofZoologyChineseAcademyofSciencesBeijing100080China statekeylaboratoryofreproductivebiology InstituteofZoologyChineseAcademyofSciencesBeijing10008

As a homodimeric glycoprotein, vascular en- dothelial growth factor (VEGF) is a highly specific mitogen of vascular endothelial cells. It can induce proliferation and migration, and inhibit apoptosis of endothelial cell. VEGF is involved in many processes in the female reproductive sys-tem, such as ovulation, periodical changes of endometrium, embryo implantation and development. VEGF plays impor-tant roles in some reproductive diseases, including pree-clampsia and fetal hypoevolutism in uterus. Based on our studies on angiogenesis and its relevant factors in the female reproductive system these years, the functions of VEGF in female reproductive system are reviewed, and the research prospect and application of VEGF are also discussed.

关键词： VEGF 血管内皮生长因子受体雌性生殖系统生物学作用血管生成生殖医学

Effect of UPP on the expression of VEGF and its receptors in mouse uterus during peri-implantation

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Chinese Science Bulletin 2003年第4期48卷 338-342页

作者： QIANDong WANGHongmei 等 statekeylaboratoryofreproductivebiology InstituteofZoologyCineseAcademyofSciencesBeijing100080China statekeylaboratoryofreproductivebiology InstituteofZoologyCineseAcademyofSciencesBeijing100080

On day 3 of gestation, one uterine horn of fe-male pregnant mouse was injected intraluminally with 5 mL 0.1 mg/mL lactacystin, a specific inhibitor of ubiquitin-pro- teasome pathway (UPP), while the contralateral horn served as control. Animals were sacrificed by cervical dislocation on day 5, 6, 7 of gestation, respectively. Then the number of implanted embryos in each uterine horn was calculated, and the expression of VEGF and its receptors was examined. The data showed that the number of implanted embryos was decreased significantly after treatment with lactacystin. The results of RT-PCR and Western blot indicated that expres-sion of VEGF and its receptors at mRNA and protein levels was significantly decreased in the treated uterus, meanwhile, the expression of HIF-1a (the a subunit of HIF, a transcrip-tional factor of VEGF) was reduced at both mRNA and pro-tein levels. These data suggested that the effect of UPP on VEGF expression was realized through regulating HIF-1a expression. In addition, UPP is likely to take part in the modulation of VEGF receptors expression. These changes may be one of the reasons for the reduction of implanted embryos.

关键词：小白鼠胚胎着植入子宫 VEGF受体表达 LIPP

Matrix metalloproteinases(MMPs)and trophoblast invasion

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Chinese Science Bulletin 2005年第12期50卷 1169-1173页

作者： LIJing ZHAOTianfu DUANEnkui statekeylaboratoryofreproductivebiology InstituteofZoologyChineseAcademyofSciencesBeijing100080China//GraduateSchooloftheChineseAcademyofSciencesBeijing100039China DepartmentofBiology BaodingTeacher'sCollegeBaoding071000China statekeylaboratoryofreproductivebiology InstituteofZoologyChineseAcademyofSciencesBeijing100080China

MMPs and their natural tissue inhibitors TIMPs are crucial in coordinated breakdown and remodel- ing of the extracellular matrix (ECM) in physiological and pathological situations. Placentation is a key event of pregnancy in which MMPs/TIMPs system plays important roles in regulating the extravillus cytotrophoblast (EVTs) invasion. This paper focuses on expression patterns and regulatory mechanisms of MMPs/TIMPs family members during the process of placentation. Their implications in curing pregnancy-related diseases are also discussed.

关键词： MMPs 细胞外矩阵组织抑制剂胚胎入侵胎盘

Expression and function of a new angiogenic factor AA98 target molecule at the maternal-embryonic boundary of rhesus monkey

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Chinese Science Bulletin 2003年第9期48卷 881-886页

作者： WEIPeng ZHANGZhihong CHENXinlei YANXiyun LIUYixun statekeylaboratoryofreproductivebiology InstituteofZoologyChineseAcademyofSciencesBeijing100080China InstituteofBiophysics ChineseAcademyofSciencesBeijing100080China

The target molecule of monoclonal antibody AA98 (AA for short) is a new vascular endothelial cell related factor and plays a role in angiogenesis as indicated by the previous data. To investigate its role in angiogenesis and placentation in primate, we examined its expression in the implantation sites on D17, 19, 28 and 34 of gestation in rhesus monkey by immunohistochemistry and Western immunoblot. Western blot analysis showed that the primary antibody used in this study was specific for its epitope. AA protein was mainly expressed in small blood vessels and in some cytotrophoblast cells. The AA staining was found mainly in the endothelial cells and vascular small *** observation supported the AA's role in angiogenesis. AA was spatio-temporarily expressed in cytotrophoblasts: weak in proliferating trophoblast within cell column and endovascular trophoblast, strong in trophoblastic subpopulation within the basal plate and vascular trophoblast; AA staining within the basal plate was down-regulated during early placentation. The shift of AA98 expression in extravillous trophoblasts suggestes a role of this new factor during the course of cytotrophoblast metastasis and spiral artery remodeling. The spatio-temporarily expression indicats that AA98 could be also used as a trophoblast cellular marker to characterize the acquisition of a vascular endothelial and invasive phenotype.

关键词： AA98 胎盘形成入侵恒河猴基因表达目标分子动物实验

Regulation of mouse blastocyst adhesion, outgrowth and secretion of matrix metalloproteinase-2 by cGMP and nitric oxide in vitro

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Chinese Science Bulletin 2004年第11期49卷 1142-1145页

作者： SHENZheng ZHAOxingxu CAOYujing BUShumin LIUWeimin DUANEnkui CollegeofVeterinaryMedicine GansuAgriculturalUniversityLanzhou730070China statekeylaboratoryofreproductivebiology InstituteofZoologyChineseAcademyofSciencesBeijing100080China

Nitric oxide (NO) is a multifunctional messen-ger molecule produced through oxidation of L-arginine to L-citrulline by enzyme NO synthase (NOS). In the current study, mouse blastocysts were cultured in the different media, and the implantation capacity of blastocyst was evaluated by evaluating the percentage of embryos adhesion and out-growth after culture for 12, 24 or 48 h. Matrix metallopro-teinase-2 (MMP-2) mRNA was detected by RT-PCR, and MMP-2 protein was detected by gelatin zymography. Inhibi-tion of blastocyst adhesion and outgrowth was observed in embryo cultured with 500 mmol/L NOS inhibitor NG-mono- methyl-L-arginine (L-NMMA) alone; however, 100 mmol/L S-nitroso-Nacetylpenicillamine (SNAP), a NO donor, and 20 mmol/L cGMP analogue, 8-Br-cGMP could block this inhibi-tion. The expression and production of MMP-2 in the blas-tocysts were suppressed by L-NMMA, and SNAP or 8-br-cGMP could reverse this suppression. These results suggest that NO induces embryo implantation by cGMP sig-naling pathway.

关键词：基体金属蛋白酶一氧化氮循环鸟苷一磷酸盐粘附生长物植入胚胎

A　Protein　Extracted　from　Mouse　Sperm　That　Plays　an　Important　Role　in　Fertilization

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Developmental and Reproductive Biology 1994年第1期3卷 1-9,T001页

作者：庄大中张天荫陈大元 statekeylaboratoryofreproductivebiology InstituteofZoologyAcademiaSinicaBeijing100080

A membrane protein was isolated from mouse sperm heads that had undergone acrosomal reaction induced by C2+ ionophore, A 23187, which, with a molecular weight of 77.6 kd, shows capability to block egg-sperm fusion. As revealed by analysis usintg isotopic markers, this protein is one of the chief membrane proteins of inner acrosomal membrane or the outer membrane of equatorial segment and Post-acrosomal region; treatment of mouse sperms with 0.6 μg/ml of the Purified protein for 30 minutes reduced the sperm-egg fusion index by 51%.The above results led us to the conclusion that the protein is an active participant in sperm-egg fusion. The possible existence of sperm receptor on egg plasma membrane was discussed.

关键词： Membrane fusion Sperm head Acrosomal reaction Egg plasma membrane Membrane protein Sperm receptor.

Infection of SARS-CoV on juvenile and adult Brandt＇s vole Microtus brandtii

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Chinese Science Bulletin 2005年第12期50卷 1199-1204页

作者： GAOHong PENGJingpian DENGWei SHIDazhao BAOLinlin WANGDehua ZHANGBinglin QINChuan ZHANGZhibin InstituteofLaboratoryAnimalsScience ChineseAcademyofMedicalSciences&PekingUnionMedicalCollegeBeijing100021China StateKeyLaboratoryofIntegratedManagementonPestInsectsandRodents StateKeyLaboratoryofReproductiveBiologyInstituteofZoologyChineseAcademyofSciencesBeijing100080China ChineseAgriculturalUniversity Beijing100094China

We studied the infectious effect of SARS-CoV virus on juvenile and adult Brandt’s Vole (Microtus brandtii) by nasal cavity spraying method (CCID50 is 105.7). SARS virus caused serious deaths in adults. The death adults demonstrated hemorrhage from mouth, nasal cavity and intestine, hemorrhageious interstitial pneumonia and gore in liver, spleen and kidney. The survival adults demonstrated local hemorrhagic spot in lung and emphysema, but the other organs showed no pathological abnormality. SARS virus caused no deaths in juveniles, but locomotion of infected juveniles became slower. In the early stage, there was local pneumonia in lung and SARS viruses were isolated from the pathological tissue. Only one control juvenile lived and the infected juvenile showed local pneumonia in lung. The results demonstrated that SARS-CoV infected Brandt’s vole seriously and adults were more susceptive to SARS-CoV than juveniles. The Brandt’s vole may be a potential animal model for SARS research.

关键词： SARS 非典型肺炎动物模型流行性传染病

Dynamic　Changes　of　β　Tubulin　during　the　Resumption　of　Meiosis　of　Mouse　Oocyte

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Developmental and Reproductive Biology 1994年第1期3卷 17-22,T001,T002页

作者：刘辉陈大元 statekeylaboratoryofreproductivebiology InstituteofZoologyAcademiaSinicaBeijing100080 StateKeyLabor

Morphological changes of tubulin during the resumption of meiosis in both mouse oocyte and fertilized egg were revealed by indirect immunofluorescent marking with monoclonal antibody against β tubulin. During germinal vesicle period (GV), tubulin was found to be distributed around the GV menibrane. With the disruption of GV membrane, microtubule complexes (MTCs) appeared in cytoplasm, first around GV membrane later to spread to other portions as well. Quantitative difference was noted among different oocytes. MTCs coexisted with spindlesformed by prometaphase tubulin, while metaphase tubulin polymerized into spindles and anaphase and telophase tubulin was concentrated in the two poles of the meiotic apparatus and the midbody. In egg arrested in the 2nd metaphase, whether maturing in vitro or vivo, all the tubulin went to form spindles with no MTCs left in the cytoplasm. After fertilization in vitro,MTCs reappeared in the egg cytoplasm activated by sperms while no MTCs could be revealed in cytoplasm after formation of pronucleus. As demonstrated by this experiment,cytoplasmic tubulin in eggs are polymerized chiefly into two forms: the star-shaped MTCs and the *** MTCs are the structure newly formed when the 1st and 2nd oocytes resumed meiosis. With colchicine disrupting the polymerization of tubulin, the maternal chromosomes,instead of orderly arrangement and orderly separation, either formed disordered mass or were divided into multiple chromatin masses. However, the penetration of sperm into egg, and decondensation and formation of pronuclei were not affected.

关键词： Oocytes Meiosis In vitro Fertilization Tubulin Micretubule complex(MTC).