Identification of egg‐jelly substances triggering sperm acrosome reaction in the newt, Cynops pyrrhogaster
Identification of egg‐jelly substances triggering sperm acrosome reaction in the newt, Cynops pyrrhogaster
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鉴定引起蝾螈精子顶体反应的卵胶物质
DOI：
10.1002/mrd.20959
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发表时间：
2009
影响因子：
2.5
通讯作者：
K. Onitake
中科院分区：
文献类型：
--
作者：
A. Watanabe;Keiko Fukutomi;H. Kubo;Manami Ohta;E. Takayama;K. Onitake
我们以前的研究表明，顶体反应（AR）发生在日本蝾螈（Cynops pyrrhogaster）的卵冻中。这与棘皮动物的物质相似，但与许多其他脊椎动物的物质不同，这些脊椎动物是从卵被膜或其衍生物--透明壳动物中衍生出来的。为了鉴定蝾螈卵胶冻中的AR诱导物质，通过筛选培养物上清液以找到最好地中和胶冻物质的AR诱导活性的单克隆抗体（mAb）来产生针对胶冻的单克隆抗体（mAb）。mAb特异性地与果冻中的蛋白条带反应。这些表观分子量为122和90 kDa的蛋白质显示出AR诱导活性，表明它们肯定是AR诱导物质。使用mAb的蛋白质印迹法表明，122和90 kDa蛋白质仅存在于卵冻的最外层，已知在该最外层发生AR诱导活性。这两种蛋白质都被麦胚凝集素（WGA）识别，WGA是一种抑制卵冻提取物中AR诱导的凝集素。综上所述，这些发现表明122和90 kDa蛋白是C.红腹菌蛋白质的WGA识别因N-糖苷酶消化而丧失，表明这些蛋白质中的N-连接碳水化合物部分可能是AR诱导活性的原因。摩尔Reprod. Dev. 76：399-406，2009.© 2008 Wiley‐利斯公司
Our previous studies have shown that the acrosome reaction (AR) occurs in egg‐jelly of the Japanese newt, Cynops pyrrhogaster. This is analogous to the substances of echinoderms but distinct from those of many other vertebrates derived from the egg envelope or its derivative, the zona pellucida. To identify the AR‐inducing substances in newt egg jelly, a monoclonal antibody (mAb) was generated against the jelly by screening the culture supernatants to find the one that best neutralized the AR‐inducing activity of the jelly substance. The mAb specifically reacted to protein bands in the jelly. These proteins, with apparent molecular weights of 122 and 90 kDa, exhibited AR‐inducing activity, indicating that they are definitely AR‐inducing substances. Western blotting using the mAb indicated that the 122 and 90 kDa proteins are present only in the egg jelly's outermost layer, where AR‐inducing activity is known to occur. Both proteins were recognized with wheat germ agglutinin (WGA), a lectin that inhibits AR‐induction in egg jelly extract. Taken together, these findings indicate that the 122 and 90 kDa proteins are the AR‐inducing substances in the egg jelly of C. pyrrhogaster. The WGA recognition of the proteins was lost by N‐glycosidase digestion, suggesting that N‐linked carbohydrate moieties in these proteins may be responsible for the AR‐inducing activity. Mol. Reprod. Dev. 76: 399–406, 2009. © 2008 Wiley‐Liss, Inc.
卵果冻的岩藻糖硫酸聚合物与精子 REJ 结合，是海胆精子顶体反应的诱导剂。
影响因子：
2.7
作者：
Vacquier,VD;Moy,GW
通讯作者：
Moy,GW