哺乳动物精卵融合必需蛋白质IZUMO1-JUNO/CD9的研究进展

精卵融合是受精过程中最为重要的步骤。目前公认IZUMO1、JUNO和CD9 (cluster of differentiation antigen 9)为精卵融合的必需蛋白质,其中IZUMO1与JUNO在精卵识别时会形成复合体。有研究表明IZUMO1、JUNO和CD9主要参与精卵融合最初的黏附过程,且它们之间是相互关联发挥作用的。近年来由于X射线晶体学相关技术的发展, IZUMO1-JUNO晶体结构基本被阐明,然而精卵融合的具体分子机制仍未被完全揭示。所以,对哺乳动物精卵融合必需蛋白质IZUMO1-JUNO/CD9的结构、功能和分子机制进行阐述是十分必要的。     

ADAMs与哺乳动物受精

ADAMs是近几年发现的一类具有多个结构区和广泛生物学功能的糖蛋白,它们在哺乳动物受精中的作用日益得到实验结果的支持,本文简要总结了近几年ADAMs在哺乳动物受精中作用的研究进展。     

ADAMs与哺乳动物受精

ADAMs是近几年发现的一类具有多个结构区和广泛生物学功能的糖蛋白,它们在哺乳动物受精中的作用日益得到实验结果的支持,本文简要总结了近几年ADAMs在哺乳动物受精中作用的研究进展。     

真菌和哺乳动物GP I锚定蛋白结构和功能差异的研究进展

GPI锚定蛋白（ Glycosylphosphatidylionsitol?anchored protein,GPI?anchored protein）是在真核生物中的一种保守的翻译后修饰蛋白,已发现有超过150种GPI锚定蛋白存在于细胞膜或细胞壁上,近年来随着对GPI锚定蛋白的深入研究,其越来越多功能被发现,我们将从GPI锚定蛋白的结构入手,详细描述其在哺乳动物和真菌中GPI锚的结构及功能的差异,以真菌的GPI锚定蛋白为切入点来研究抗真菌药物但同时要留意该类药物对构成人体细胞的GPI锚定蛋白的潜在性损伤,为将来抗真菌药物的研发提供新的思路。     

哺乳动物精卵相互作用的分子机制的研究进展

哺乳动物受精是由一系列有序步骤组成的复杂细胞相互作用过程组成,最终导致精卵质膜融合形成受精卵。近来运用基因组学和蛋白质组学技术在哺乳动物精子和卵子表面鉴定出若干可能参与精卵质膜粘附与融合过程的蛋白分子,并对其结构和生物学功能进行了研究,但精卵识别与融合的分子机理仍然不清楚。综述了与精卵识别和融合有关的蛋白的最新研究进展,为进一步研究精卵相互作用的分子机制提供参考。     

哺乳动物X染色体失活机制

哺乳动物X染色体连锁基因的剂量平衡,是通过雌性胚胎发育早期随机或印记失活一条X染色体来实现的,这是一个复杂的过程,包括：启动、计数、选择、维持等一系列的步骤。X染色体失活中心是X染色体失活的主控开关座位,调节X失活的早期事件,失活发生后,X染色体的失活状态可稳定地存在并传递给后代,这一过程涉及基因组印记的形成。此外,在雄性动物,精原细胞减数分裂早期也存在着短暂的X染色体失活现象。现对哺乳动物X染色体失活机制的最新进展进行综述。     

哺乳动物性别决定的分子要素

哺乳动物性别分化是由胚胎时期性腺分泌的激素控制,而在激素分泌前,ＸＸ和ＸＹ胎儿都具有牟勒氏管（Ｍüｌｅｒｉａｎｄｕｃｔ）,吴夫氏管（Ｗｏｉｆｉａｎｄｕｃｔ）两套生殖导管和未分化的性腺。牟勒氏管将来分化为输卵管,子宫等雌性生殖道,吴夫氏管则分化为输精管...     

哺乳动物性别决定机制的研究进展

哺乳动物的性别决定包括初级性别决定和次级性别决定,是以SRY基因为主导,其他多个基因参与的级联调控过程。近年的研究表明。SRY、DAX1、SOX3等性染色体基因和SOX9、MIS、WT1、SF1等常染色体基因都参与性别决定的级联过程。结合中学生物学教材及发育生物学相关原理,从性染色体上和常染色体上与性别决定有关的基因阐述哺乳动物的性别决定机制,并简述了哺乳动物的性别决定模型。     

哺乳动物性别决定的研究进展

近年来,人们对与哺乳动物性别决定相关的SRY、SOX9、SF－1、WT1和DAX－1基因的结构、功能和产物之间的相互作用进行了一系列的研究,使人们对哺乳动物的性别决定分子机制的探索又向前推进了一步,这将对发育生物学和性别决定的进化研究起到了推动作用。     

哺乳动物胚胎滞育及其调控机制研究进展

胚胎滞育(Embryonic diapause)是存在于部分哺乳动物中的一种生存策略和繁殖状态.其具体过程为:胚胎在着床前停止或减缓发育形成胚胎滞育期,滞育期结束后胚胎再次活化,且滞育过程不会对随后的胚胎发育产生任何不良影响.胚胎滞育主要分为两种类型:兼性滞育(Facultative diapause)和专性滞育(Ob...     

Infertility of CD9-deficient mouse eggs is reversed by mouse CD9, human CD9, or mouse CD81; polyadenylated mRNA injection developed for molecular analysis of sperm-egg fusion

CD9 is a membrane protein belonging to the tetraspanin family. Despite CD9's broad tissue distribution, the only abnormality observed in CD9-deficient mice was infertility of females, which was responsible for a defect in the sperm-egg fusion process. However, the function of CD9 in sperm-egg fusion is not clear at all because the technique to analyze the activity of molecules in sperm-egg fusion has not been established. We demonstrated that the exogenous mouse CD9, expressed by polyadenylated mRNA injection at the germinal-vesicle stage oocytes, was precisely localized to the egg plasma membrane, and the expression reversed the infertility of CD9(-/-) eggs. Then, two other tetraspanins, human CD9 and mouse CD81, overexpressed with this technique on CD9(-/-) eggs restored the fertilization rate up to approximately 90 and approximately 50% against that of wild type eggs, respectively. Moreover, in the presence of an anti-mouse CD9 mAb, which blocks sperm-egg fusion, expression of human CD9 or mouse CD81 on eggs also rescued the fusibility. These results suggested that human CD9 plays a crucial role in human fertilization, and mouse CD81 has the potential to compensate for CD9 function in sperm-egg fusion. In addition, the polyadenylated mRNA injection is effective for molecular analysis of sperm-egg fusion.     

None of the integrins known to be present on the mouse egg or to be ADAM receptors are essential for sperm-egg binding and fusion

Antibody inhibition and alpha6beta1 ligand binding experiments indicate that the egg integrin alpha6beta1 functions as a receptor for sperm during gamete fusion; yet, eggs null for the alpha6 integrin exhibit normal fertilization. Alternative integrins may be involved in sperm-egg binding and fusion and could compensate for the absence of alpha6beta1. Various beta1 integrins and alphav integrins are present on mouse eggs. Some of these integrins are also reported to be receptors for ADAMs, which are expressed on sperm. Using alpha3 integrin null eggs, we found that the alpha3beta1 integrin was not essential for sperm-egg binding and fusion. Oocyte-specific, beta1 integrin conditional knockout mice allowed us to obtain mature eggs lacking all beta1 integrins. We found that the beta1 integrin null eggs were fully functional in fertilization both in vivo and in vitro. Furthermore, neither anti-mouse beta3 integrin function-blocking monoclonal antibody (mAb) nor alphav integrin function-blocking mAb inhibited sperm binding to or fusion with beta1 integrin null eggs. Thus, function of beta3 or alphav integrins does not seem to be involved in compensating for the absence of beta1 integrins. These results indicate that none of the integrins known to be present on mouse eggs or to be ADAM receptors are essential for sperm-egg binding/fusion, and thus, egg integrins may not play the role in gamete fusion previously attributed to them.     

精卵质膜融合过程中卵膜上的候选分子

受精过程中精卵质膜融合分子机制的研究一直备受关注。随着基因剔除技术的发展及众多新技术的应用,人们发现这一过程涉及多种分子,目前对卵膜上的CD9、糖基化磷脂酰肌醇锚定蛋白和整合素研究较多。现从细胞层次和分子层次上总结三者在精卵质膜融合方面的实验及结论,分析各个实验结果矛盾之处,讨论精卵质膜融合研究的前景。     

Structure,regulatory factors and cancer-related physiological effects of ADAM9

ABSTRACT

The ADAMs family belongs to the transmembrane protein superfamily of zinc-dependent metalloproteases, which consists of multiple domains. These domains have independent but complementary functions that enable them to participate in multiple biological processes. Among them, ADAM9 can not only participate in the degradation of extracellular matrix as a metalloprotease, but also mediate tumor cell adhesion through its deintegrin domain, which is closely related to tumor invasion and metastasis. It is widely expressed in a variety of tumor cells and can affect the proliferation, invasion and metastasis of related cancer cells. We provide our views on current progress, its increasing importance as a strategic treatment goal, and our vision for the future of ADAM9.     

EWI-2 regulates alpha3beta1 integrin-dependent cell functions on laminin-5

EWI-2, a cell surface immunoglobulin SF protein of unknown function, associates with tetraspanins CD9 and CD81 with high stoichiometry. Overexpression of EWI-2 in A431 epidermoid carcinoma cells did not alter cell adhesion or spreading on laminin-5, and had no effect on reaggregation of cells plated on collagen I (alpha2beta1 integrin ligand). However, on laminin-5 (alpha3beta1 integrin ligand), A431 cell reaggregation and motility functions were markedly impaired. Immunodepletion and reexpression experiments revealed that tetraspanins CD9 and CD81 physically link EWI-2 to alpha3beta1 integrin, but not to other integrins. CD81 also controlled EWI-2 maturation and cell surface localization. EWI-2 overexpression not only suppressed cell migration, but also redirected CD81 to cell filopodia and enhanced alpha3beta1-CD81 complex formation. In contrast, an EWI-2 chimeric mutant failed to suppress cell migration, redirect CD81 to filopodia, or enhance alpha3beta1-CD81 complex formation. These results show how laterally associated EWI-2 might regulate alpha3beta1 function in disease and development, and demonstrate how tetraspanin proteins can assemble multiple nontetraspanin proteins into functional complexes.     

Expression, purification, and refolding of mouse islet neogenesis associated protein-related protein for NMR studies

Islet neogenesis associated protein-related protein (INGAPrP) is thought to be involved in the differentiation of non-insulin-producing cells to insulin-secreting cells. INGAPrP is a mouse gene product that has a 72% identical amino acid sequence to a known islet-generating factor, hamster islet-neogenesis-associated protein (INGAP), which acts by differentiating pancreatic ductal cells into beta-cells. The three-dimensional structure of these proteins is unknown. The structure would provide information about the conformation of the active portion of INGAP, the so-called INGAP pentadecapeptide, leading to a well-defined target for rational drug design. An efficient procedure for the production of INGAPrP would facilitate the process of structure determination. We have successfully produced and isolated (15)N-labeled INGAPrP by expression in Escherichia coli Rosetta (DE3) cells in Spectra-9 media followed by a two-step purification and refolding protocol. The hexahistidine tag engineered at the N-terminus of the protein is used in the first step for standard immobilized-metal affinity chromatography purification under denaturing conditions. The secondary purification step utilizes a gel permeation chromatography column, producing homogeneous INGAPrP as well as refolding the protein. To verify that the protein was folded, we performed a (1)H-(15)N HSQC NMR experiment that showed excellent dispersion of signals, indicative of a folded protein. We also performed circular dichroism experiments, which demonstrated the presence of secondary structure. In summary, we report the first expression and isolation of INGAPrP, as well as demonstrate that our method produces a folded protein, which is necessary for structure determination.