首页> 美国卫生研究院文献>Biology of Reproduction >WBP2 shares a common location in mouse spermatozoa with WBP2NL/PAWP and like its descendent is a candidate mouse oocyte-activating factor
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WBP2 shares a common location in mouse spermatozoa with WBP2NL/PAWP and like its descendent is a candidate mouse oocyte-activating factor

机译:WBP2与WBP2NL / PAWP在小鼠精子中共有一个共同的位置并且其后代是候选的小鼠卵母细胞激活因子

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摘要

The sperm-borne oocyte-activating factor (SOAF) resides in the sperm perinuclear theca (PT). A consensus has been reached that SOAF most likely resides in the postacrosomal sheath (PAS), which is the first region of the PT to solubilize upon sperm–oocyte fusion. There are two SOAF candidates under consideration: PLCZ1 and WBP2NL. A mouse gene germline ablation of the latter showed that mice remain fertile with no observable phenotype despite the fact that a competitive inhibitor of WBP2NL, derived from its PPXY motif, blocks oocyte activation when coinjected with WBP2NL or spermatozoa. This suggested that the ortholog of WBP2NL, WBP2, containing the same domain and motifs associated with WBP2NL function, might compensate for its deficiency in oocyte activation. Our objectives were to examine whether WBP2 meets the developmental criteria established for SOAF and whether it has oocyte-activating potential. Immunoblotting detected WBP2 in mice testis and sperm and immunofluorescence localized WBP2 to the PAS and perforatorium of the PT. Immunohistochemistry of the testes revealed that WBP2 reactivity was highest in round spermatids and immunofluorescence detected WBP2 in the cytoplasmic lobe of elongating spermatids and colocalized it with the microtubular manchette during PT assembly. Microinjection of the recombinant forms of WBP2 and WBP2NL into metaphase II mouse oocytes resulted in comparable rates of oocyte activation. This study shows that WBP2 shares a similar testicular developmental pattern and location with WBP2NL and a shared ability to activate the oocyte, supporting its consideration as a mouse SOAF component that can compensate for a WBP2NL.
机译:精子携带的卵母细胞激活因子(SOAF)驻留在精子核周囊(PT)中。已经达成共识,SOAF最有可能驻留在顶体后鞘(PAS)中,这是PT在精卵融合后可溶解的第一个区域。正在考虑两个SOAF候选对象:PLCZ1和WBP2NL。尽管后者的PPXY基序产生的竞争性WBP2NL抑制剂在与WBP2NL或精子同时注射时阻止了卵母细胞的活化,但后者的小鼠基因种系消融显示小鼠仍可育,没有可观察的表型。这表明WBP2NL的直系同源基因WBP2包含与WBP2NL功能相关的相同结构域和基序,可能弥补了其在卵母细胞激活中的不足。我们的目标是检查WBP2是否符合SOAF确立的发育标准,以及是否具有卵母细胞激活潜能。免疫印迹法在小鼠睾丸和精子中检测到WBP2,免疫荧光将WBP2定位于PT的PAS和穿孔。睾丸的免疫组织化学显示,圆形精子中WBP2反应性最高,免疫荧光检测到伸长的精子的细胞质叶中WBP2并在PT组装过程中与微管手枪共定位。将WBP2和WBP2NL的重组形式显微注射到中期II小鼠卵母细胞中导致卵母细胞活化的速率相当。这项研究表明,WBP2与WBP2NL具有相似的睾丸发育模式和位置​​,并具有激活卵母细胞的共同能力,从而支持将其视为可以补偿WBP2NL的小鼠SOAF组分。

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