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试验旨在探讨不同发育时期卵泡上四跨膜蛋白CD9的表达,以及体外成熟和超低温冷冻对绵羊卵母细胞CD9的影响。采用免疫组织化学技术检测CD9蛋白在卵泡上的表达部位,实时荧光定量PCR技术检测CD9 mRNA的表达量,Western blotting技术检测CD9蛋白的含量。免疫组织化学结果发现,在绵羊原始卵泡上就开始检测到CD9荧光信号,且随着卵泡的成熟荧光信号逐渐增强,成熟卵泡时荧光信号达到最强;实时荧光定量PCR检测新鲜MⅡ期卵母细胞中CD9 mRNA的表达量显著增高（P<0.05）,冷冻GV期卵母细胞CD9 mRNA的表达量最少;Western blotting检测得出新鲜MⅡ期卵母细胞中CD9蛋白表达最多,冷冻GV期卵母细胞上表达最少,与实时荧光定量PCR结果一致。卵母细胞的冷冻保存要比胚胎保存应用前景广泛,而四跨膜蛋白CD9在精卵融合中更有着重要的作用,以上结果均显示,冷冻保存降低卵母细胞四跨膜蛋白CD9含量,而CD9的损伤是受精率下降的重要原因之一。 相似文献
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【目的】研究谷胱甘肽过氧化酶5(GPx5)基因在绵羊附睾及附睾不同发育时期的表达特性。【方法】采用荧光定量PCR技术、免疫印迹技术及免疫荧光等方法,分别检测GPx5基因在绵羊附睾不同发育时期、不同部位上mRNA和蛋白的表达及分布情况。【结果】荧光定量PCR结果表明,青年羊附睾GPx5基因在头部的mRNA表达量显著高于体部和尾部(P0.05);青年羊附睾头部GPx5基因mRNA表达量显著高于羔羊和老年羊附睾头部(P0.05)。免疫印迹结果表明,羔羊附睾无GPx5蛋白表达;青年羊附睾各部位均有表达,而老年羊附睾头部和体部有表达、尾部无表达。灰度分析结果表明,青年羊附睾头部GPx5相对表达量显著高于其他部位(P0.05),且青年羊附睾头部GPx5蛋白相对表达量显著高于老年羊附睾头部(P0.05)。免疫荧光结果表明,在羔羊附睾未检测到GPx5蛋白表达信号;青年羊附睾各部位均有荧光信号表达,且附睾头部和体部荧光强度明显高于尾部,并在附睾管液体中也发现有荧光信号;老年羊附睾头部和体部有荧光信号,荧光遍布于附睾管假复层上皮细胞的细胞核及细胞质中。【结论】GPx5基因mRNA在绵羊附睾不同发育时期均有表达,但在不同部位具有表达差异性;GPx5蛋白于附睾管假复层上皮细胞表达,且在性成熟前羔羊附睾及老年羊附睾尾部未表达,因此推测GPx5蛋白的表达与绵羊性发育时期有密切关联。 相似文献
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SHI Zhen-dan Wuliyasu Wuyahan Ailungaowa MO Li-mei Zhamuga Wuyunsiqin ZHANG Tie-jia Narenhua 《中国畜牧兽医》2016,43(5):1277-1284
Tetraspanins with special four-times acrossing membrane structure can contact outer, intrinsic membrane and transmembrane proteins, which forms a channel to connect the internal and external membrane signal.CD81, as one member of Tetraspanins, has a variety of biological activities and is widely dispersed and participates in the organism physiological responses.CD81 plays an important role in many cell process, and is closely related to the pathogenesis of many human diseases, and so widely watched.But the molecular structure, space conformation and physiological mechanisms of CD81 are still not entirely clear.We still need to do further research and discussion, especially, the comprehensively understanding of precise role of CD81 in the process of HCV into a host cell, it is vital for clinical application.This article would focus on the research progresses of molecular structure and biochemical characteristics, cell fusion effect, the virus-identified mechanism, the role in the immune system and clinical application on CD81.In summary, clarifing the function and role of CD81 is important to study the clinical pathogenesis and treatment of various diseases. 相似文献
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Tetraspanins特殊的四次跨膜结构能联系膜内、跨膜和膜外蛋白,从而形成一个通道,起到连接膜内外生物信号的作用。CD81是一种具有多种生物学活性的四次跨膜蛋白分子,在机体内分布广泛并参与生物体大量的生理应答反应,在多个细胞过程中充当重要角色,且与人类很多疾病的发病机理息息相关,受到广泛关注并成为研究热点。目前对CD81的分子结构、空间构象及在不同组织细胞中的作用机制尚未完全清楚,仍有待于进一步研究与探讨,尤其是探究CD81在丙型肝炎病毒(HCV)感染宿主细胞过程中的精确作用对于临床应用至关重要。作者将重点阐述近年在CD81的分子结构与生化特征、细胞融合效应、识别病毒的机制、在免疫系统中的作用、临床应用等研究方面取得的进展和意义,这对研究临床各种疾病的发病机理和治疗都具有重要的现实意义。 相似文献
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Wuliyasu Wuyahan SHI Zhen-dan Ailungaowa Zhamuga Wuyunsiqin Enhemarite Narenhua 《中国畜牧兽医》2016,43(8):2104-2111
The experiment was aimed to explore the expression of CD9 in the follicles at different developmental stages, and investigate the effect of in vitro maturation and cryopreservation on CD9. The expression of CD9 protein in the follicle was detected by immunohistochemistry technique, the expression of CD9 mRNA was detected by Real-time quantitative PCR, CD9 protein was detected by Western blotting. The results showed that the CD9 fluorescence signal was detected by immunohistochemistry technique, and the fluorescence signal gradually increased with the maturation of the follicle, and fluorescence signal was the strongest in mature follicle;Real-time quantitative PCR showed that the expression of CD9 mRNA in fresh MⅡ oocytes was significantly increase (P<0.05), and was the least in frozen GV oocytes;The result of Western blotting indicated that CD9 protein was the most in fresh MⅡ oocytes and the expression of frozen GV oocytes was the least, consistent with the results of Real-time quantitative PCR. Cryopreservation of oocytes was more extensive than embryo cryopreservation. Tetraspanins CD9 played a very important role in sperm egg fusion. The above experimental results showed that the cryopreservation made damage to oocytes, decrease content of tetraspanins CD9 protein, and CD9 injury was one of the important reasons for the decline of fertilization rate. 相似文献
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