[1]高立芳,赵邦霞,谭冬梅,等.瘦素在着床前小鼠胚胎中的表达及其对胚胎发育的影响[J].第三军医大学学报,2008,30(14):1316-1318.
 GAO Li-fang,ZHAO Bang-xia,TAN Dong-mei,et al.Leptin expression in mouse pre-implantation embryos and its effect on embryo development in vitro[J].J Third Mil Med Univ,2008,30(14):1316-1318.
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瘦素在着床前小鼠胚胎中的表达及其对胚胎发育的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第14期
页码:
1316-1318
栏目:
论著
出版日期:
2008-07-30

文章信息/Info

Title:
Leptin expression in mouse pre-implantation embryos and its effect on embryo development in vitro
作者:
高立芳赵邦霞谭冬梅何明忠谭毅
重庆医科大学实验动物中心
Author(s):
GAO Li-fang ZHAO Bang-xia TAN Dong-mei HE Ming-zhong TAN Yi
Laboratory Animal Center, Chongqing Medical University, Chongqing 400016, China
关键词:
瘦素胚胎发育免疫荧光
Keywords:
leptin embryo development RT-PCR immunofluorescence
分类号:
Q95-33;Q954.4
文献标志码:
A
摘要:
目的  探讨瘦素(leptin)在围着床期小鼠胚胎中的表达规律和分布情况以及leptin抗体对小鼠胚胎体外发育的影响。 方法  雌性NIH小鼠超排后与雄鼠交配,分别在妊娠1~4 d上午从输卵管或子宫角冲取相应时期的胚胎,用于以下实验:①分别提取总RNA,检测胚胎中leptin mRNA表达情况。②采用免疫荧光技术,观察leptin蛋白在囊胚的表达和分布情况。③将8-细胞小鼠胚胎培养于含不同浓度leptin抗体的培养液中,观察囊胚形成及脱透明带的情况。 结果  leptin mRNA仅在囊胚期特异性表达,其余各时期胚胎未见表达;leptin蛋白在囊胚的滋养层细胞和内细胞团的细胞质和细胞膜呈阳性表达,而在细胞核无表达。一定浓度的leptin抗体可明显降低囊胚的形成率(1∶400, P<0.05) 和脱带率(1 ∶1 600, P<0.05),且随着抗体浓度的增加,这种抑制作用更明显。 结论  leptin能促进着床前胚胎发育,同时也可能参与了胚胎着床过程。
Abstract:
Objective    To investigate the expression and distribution of leptin in pre-implantation embryo, and its effect on mouse embryo development in vitro.     Methods    Adult NIH female mice were superovulated and then mated with fertile males of the same strain. On the morning of 1st to 4th d of pregnancy, the oviduct and horn of uterus were dissected out and flushed with DMEM/F12 medium adequately. The embryos collected were subjected to the following procedures: ①Total RNA was isolated from embryos of each stage respectively and mRNA expression of leptin was detected by RT-PCR. ②Immunofluorescence staining was performed on blastocysts to analyze the expression and the distribution of leptin protein under laser confocal microscope.  ③Eight-cell embryos were cultured in medium containing leptin antibody at various concentrations to observe the formation and hatching of blastocysts.     Results    ①Leptin mRNA expression was only detected in blastocyst, and no expression was detected in embryos in other stages. Leptin protein was detected in cytoplasma, membrane of trophoblastic cell and inner cell mass, however no expression was detected in cell nucleus.  ②Leptin polyclonal antibody significantly inhibited formation of blastocysts (1∶400, P<0.05) and prevented hatching of blastocysts from zona pellucida (1∶1 600, P<0.05) in a dose-dependent manner.     Conclusion    The temporal expression of leptin suggests that leptin might play an important role during embryo development and implantation process in mice.

参考文献/References:

高立芳,赵邦霞,谭冬梅,等.  瘦素在着床前小鼠胚胎中的表达及其对胚胎发育的影响[J].第三军医大学学报,2008,30(14):1316-1318.

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更新日期/Last Update: 2008-07-17