Lakoski K A, Carron C P, Cabot C L, Saling P M
Department of Zoology, Duke University, Durham, North Carolina 27710.
Biol Reprod. 1988 Feb;38(1):221-33. doi: 10.1095/biolreprod38.1.221.
Development of the sperm's capacity to interact with the zona pellucida was investigated at the stage when the acrosome reaction (AR) is induced. The response of epididymal sperm to agents that affect the occurrence of the AR was used to monitor maturational changes. Despite the finding that sperm from the three main epididymal regions were competent to undergo ARs induced by the divalent cation ionophore A23187 (56% AR, 74% AR, and 83% AR in caput, corpus, and cauda, respectively), the cells' responses to solubilized zonae pellucidae were different. When challenged with 5 zonae equivalents/microliter, both corpus and cauda sperm shed their acrosomes in high numbers (75% AR and 86% AR, respectively), whereas caput sperm did not (23% AR). Previous work has shown that the presence of M42 monoclonal antibody (mAb) during in vitro and in vivo fertilization inhibits sperm penetration through the zona pellucida by specific interference with zonae pellucidae-induced ARs. In this study, presence of the M42 mAb did not affect the incidence of A23187-induced ARs, whereas the zona-induced ARs that occurred in both corpus and cauda sperm were inhibited fully with M42 immunoglobulin (Ig) G. In addition, the antigen recognized by M42 mAb on sperm, termed M42 Ag, was examined during epididymal maturation. Although antigen localization appeared indistinguishable by immunofluorescence on sperm taken from the caput, corpus, and cauda regions of the epididymis, modification of this antigen during epididymal transit was detected. Equilibrium-binding studies using 125I-M42 IgG demonstrated a progressive increase during epididymal transit in the amount of M42 mAb that bound to fixed cells. Corpus and cauda sperm bound 185% and 240%, respectively, of the 125I-M42 IgG detected on caput sperm. These changes in expression of M42 Ag paralleled a structural change: the Mr of the antigen decreased from a 195,000/210,000 doublet in caput sperm to a 185,000/200,000 doublet in corpus and cauda sperm, as determined by immunoblot analysis of sodium dodecyl sulfate (SDS)-extracted sperm. Results presented here demonstrate that mouse sperm develop the capacity to undergo a zona-induced AR during epididymal maturation. The M42 antigen, which is involved in the zona-induced AR, is modified during epididymal transit coincident with development of the sperm's responsiveness to zonae. Our working hypothesis, based on these results, is that development of the sperm's capacity to undergo a physiological AR is related to modification of M42 Ag.
在诱导顶体反应(AR)的阶段,研究了精子与透明带相互作用能力的发育情况。利用附睾精子对影响AR发生的试剂的反应来监测成熟变化。尽管发现来自附睾三个主要区域的精子都能够经历由二价阳离子载体A23187诱导的AR(分别在附睾头、体和尾中的AR发生率为56%、74%和83%),但细胞对溶解的透明带的反应是不同的。当用5个透明带当量/微升进行刺激时,附睾体和尾的精子大量顶体脱落(分别为75%和86%的AR),而附睾头的精子则没有(23%的AR)。先前的研究表明,在体外和体内受精过程中,M42单克隆抗体(mAb)的存在通过特异性干扰透明带诱导的AR来抑制精子穿透透明带。在本研究中,M42 mAb的存在不影响A23187诱导的AR的发生率,而附睾体和尾精子中发生的透明带诱导的AR被M42免疫球蛋白(Ig)G完全抑制。此外,在附睾成熟过程中,检测了精子上被M42 mAb识别的抗原,即M42 Ag。尽管通过免疫荧光对取自附睾头、体和尾区域的精子进行检测时,抗原定位看起来没有区别,但在附睾转运过程中检测到了该抗原的修饰。使用125I-M42 IgG进行的平衡结合研究表明,在附睾转运过程中,与固定细胞结合的M42 mAb的量逐渐增加。附睾体和尾的精子分别结合了在附睾头精子上检测到的125I-M42 IgG的185%和240%。M42 Ag表达的这些变化与一种结构变化平行:通过对十二烷基硫酸钠(SDS)提取的精子进行免疫印迹分析确定,该抗原的相对分子质量从附睾头精子中的195,000/210,000双峰降低到附睾体和尾精子中的185,000/200,000双峰。这里给出的结果表明,小鼠精子在附睾成熟过程中发展出了经历透明带诱导的AR的能力。参与透明带诱导的AR的M42抗原在附睾转运过程中与精子对透明带反应性的发展同时发生修饰。基于这些结果,我们的工作假设是,精子经历生理性AR的能力的发展与M42 Ag的修饰有关。