β-microseminoprotein in human spermatozoa and its potential role in male fertility
β-microseminoprotein in human spermatozoa and its potential role in male fertility
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DOI：
10.1530/rep-08-0032
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发表时间：
2008-08-01
期刊：
影响因子：
3.8
通讯作者：
Coronel, Carlos E.
中科院分区：
文献类型：
--
作者：
Franchi, N. Anahi;Avendano, Conrado;Coronel, Carlos E.
β-微精浆蛋白（MSMB）是人类精浆中最丰富的蛋白质之一。本研究的目的是：（1）从精浆（SP）中纯化MSMB并制备MSMB抗体;（2）研究MSMB与射精精子的相互作用及其对自发顶体反应（AR）的可能影响;（3）定量SP中MSMB的含量并研究其与精子临床参数的关系。从SP中纯化MSMB，并使用特异性多克隆抗体通过间接免疫荧光检查其在精子表面上的存在。以豚鼠附睾精子为模型，观察MSMB对AR的影响。使用两种针对MSMB的多克隆抗体，通过双位点结合ELISA进行MSMB定量测定。根据世界卫生组织的标准，在来自生育供体（对照）和生育力低下患者的精液样本中评估MSMB。MSMB主要分布在精子的头部和颈部的顶体区域。用MSMB预孵育豚鼠附睾精子，观察到显著的自发AR抑制。最后，与生育对照组相比，生育力低下患者的MSMB显著增加（P
beta-Microseminoprotein (MSMB) is one of the most abundant proteins in human seminal plasma. The objectives of this study were: (1) to purify MSMB from seminal plasma (SP) and generate antibodies against the pure protein; (2) to investigate the interaction of MSMB with ejaculated spermatozoa and its possible effect on the spontaneous acrosome reaction (AR); and (3) to quantify MSMB content in SP and examine its relationship with the clinical sperm parameters. MSMB was purified from SP and its presence on the sperm surface was examined by indirect immunofluorescence using a specific polyclonal antibody. The effect of MSMB on the AR was evaluated using guinea pig epididymal spermatozoa as a model. MSMB quantification assay was performed with a two-site binding ELISA using two polyclonal antibodies against MSMB. MSMB was assessed in semen samples from fertile donors (controls) and subfertile patients according to World Health Organization criteria. MSMB was detected on the sperm surface and mainly localized to the acrosomal region of the head and neck. A significant spontaneous AR inhibition was observed when guinea pig epididymal spermatozoa were preincubated with MSMB. Finally, MSMB was significantly increased in subfertile patients when compared with fertile controls (P