【作者】 吴吉文；

【导师】 王玮； 木全弘治；

【作者基本信息】 福建医科大学 ， 人体解剖学与组织胚胎学， 2005， 硕士

【摘要】 目的观察小鼠卵丘细胞外基质透明质酸(hyaluronic acid, HA)和透明质酸结合蛋白(HA-binding proteins ,HABPs)的分布,探讨透明质酸结合蛋白对小鼠卵丘的结构和功能的影响。方法取3-4 周龄的雌鼠(含野生型和bikunin 基因敲除小鼠)诱导排卵后取卵丘-卵母细胞复合体(cumulus oocyte complex,COC)。实验动物分为野生型组和bikunin 基因敲除小鼠组各5 只,诱导排卵后镜下计数卵母细胞。免疫荧光双重染色后观察HA 及HABPs,如SHAP(serum-derived hyaluronan-associated proteined),versican,LP(Linkprptein),CD44 和TSG-6(tumor necrosis factor-stimulated gene-6),在卵丘细胞外基质的分布。结果透明质酸和透明质酸结合蛋白在小鼠卵丘细胞外基质呈阳性表达,透明质酸结合蛋白的缺失损害卵丘细胞外基质完整。HA 和SHAP 共同分布在野生型小鼠卵丘细胞外基质。在bikunin 基因敲除小鼠卵丘细胞外基质,SHAP 未发现,HA 仍分布在卵丘细胞之间,但卵丘细胞之间的网格状消失,提示SHAP-HA 复合体缺失造成卵丘细胞外基质的破毁。免疫荧光双重染色显示野生型小鼠COC 中CD44,LP,TSG-6 和HA 的存在。CD44 分布在野生型小鼠卵丘细胞膜表面,SHAP,LP,TSG-6 和HA 共同分布在野生型小鼠卵丘细胞外基质。versican 表达并和透明质酸分布于卵丘细胞外基质,bikunin 基因并不能影响versican 的表达。SHAP-HA 复合体缺失影响卵母细胞的输送。bikunin 基因敲除小鼠和正常对照组的卵母细胞数有显著区别(P<0.01)。在bikunin 基因敲除小鼠,更多的卵母细胞滞留在卵巢和输卵管的囊泡间。更多还原

【Abstract】 ObjectiveTo study the effects of hyaluronan(HA) and hyaluronan-associatedproteins(HABPs) on the cumulus extracellular matrix in mice. and to investigatethe effect of HABPs on the the structure and function of the cumulus oophorus.MethodsThree-to four-week-old female mice(wild-type and bikunin-null mice) wereinstimulated to superovulation and isolation of COC, the cumulus oocytescomplexes were counted with microscope. Immunofluoresence staining was used todetect the distribution of HA and HABPs in the cumulus extracellular matrix.ResultsHA and HABPs-positive staining were presented in the cumulus extacellularmatrix. The deficiency of the HABPs impairs the integrity of cumulusextracellular matrix. SHAP and HA were distributed in the cumulus matrix of thewild-type mice. SHAP was not detected in the cumulus matrix of the bikunin-nullmice. HA was distributed between the cumulus cells, but the lattice of the cumuluscells was dismissed, showed that SHAP-HA complex-deficiency cause abnormalityof the cumulus matrix. Immunofluoresence staining indicated HA、LP、CD44 andTSG-6 were presented in the cumulus extacellular matrix. versican was localizedto the expanded cumulus matrix as well as HA, versican expression andlocalization were not altered in the bikunin-knockout mice. SHAP-HAcomplex-deficiency affect the transport of the oocytes. The oocyte countingshows the induced ovulation, there is a significant difference between thebikunin-null mice and the normal controls. In bikunin-null mice, more oocyteswere found in the bursal space between ovary and oviductConclusion SHAP、versican、LP、CD44 and TSG-6 are required for the construction ofintercellular matrices. HABPs is required for the of integrity intercellularmatrices .The function of the oocytes were reduced significantly for themalformation of the matrices.更多还原