畜牧兽医学报2017，Vol.48Issue(4)：652-659,8.DOI:10.11843/j.issn.0366-6964.2017.04.008
小鼠精子透明质酸酶SPAM1在受精过程中的功能研究
Functional Analysis of Mouse Sperm Hyaluronidase SPAM1 in Fertilization
摘要
Abstract
This study was aimed to elucidate the possible functions of sperm adhesion molecule 1 (SPAM1) on the sperm/cumulus interaction during fertilization and explore its potential mechanism.Genomic DNA was extracted from the tails of mice and genotype for Spam1 null mutation was confirmed by PCR analysis.The protein of the screened wild type (WT) and Spam1-deficient (KO) sperm were extracted and confirmed by Western blot,and the hyaluronidase activity was measured by the colorimetric method.The epididymal sperm were capacitated by incubation for 2 h and subjected to the assays of sperm motility,sperm/cumulus interaction or in vitro fertilization (IVF).These results showed that the SPAM1 protein was readily absent in the KO sperm,and the hyaluronic acid-hydrolyzing enzymes activity was significantly reduced in the KO sperm,compared to that of the WT sperm (P＜0.01).There were no significant differences in sperm motility between the KO and WT groups after capacitation (P＞0.05).However,the loss of SPAM1 significantly influenced the percentage of acrosome reacted sperm in the extracellular matrix of the cumulus (P＜0.01),resulted in a markedly decrease in sperm entry into and/or penetration of the cumulus mass,only a few sperm succeeded in access to the surface of the oocyte zona pellucida (ZP) and contrastly a remarkably great deal of sperm accumulated on the surface or outer edge of the cumulus (P＜0.01).In addition,sperm lacking SPAM1 caused the delayed cumulus disperse and fertilization at IVF 2 h (P＜0.05).Therefore,mouse sperm hyaluronidase SPAM1 may be associated with the acrosome reaction and affect the sperm/cumulus interaction,which reveal that SPAM1 may play other possible roles in sperm access to the oocyte ZP except the hyaluronan-degrading activity.关键词
SPAM1/精子/卵丘互作/顶体反应/精子/小鼠Key words
SPAM1/sperm/cumulus interaction/acrosome reaction/sperm/mouse分类
生物科学引用本文复制引用
周崇,黄莉,石德顺,蒋建荣,马场忠..小鼠精子透明质酸酶SPAM1在受精过程中的功能研究[J].畜牧兽医学报,2017,48(4):652-659,8.基金项目
广西自然科学基金(2014GXNSFCA118008) （2014GXNSFCA118008）
