Determination of the capacity of ram epididymal and ejaculated sperm to undergo the acrosome reaction and penetrate ova.
Determination of the capacity of ram epididymal and ejaculated sperm to undergo the acrosome reaction and penetrate ova.
复制标题
测定公羊附睾和射精精子进行顶体反应和穿透卵子的能力。
DOI：
10.1095/biolreprod44.6.1080
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发表时间：
1991
影响因子：
3.6
通讯作者：
Hammerstedt,RH
中科院分区：
文献类型：
--
作者：
Williams,RM;Graham,JK;Hammerstedt,RH
了解精子的附睾成熟需要描述膜特性的变化及其与细胞功能变化的关系。虽然精子膜已经在公羊中进行了一些详细的研究，但很少有报告涉及相关的功能变化。本报告通过评价（a）每个附睾区域细胞的精子顶体反应（AR）诱导的时间过程;（B）附睾精子穿透卵子的能力;（c）生理AR和一般精子变性的差异;和（d）附睾精子的顶体酶释放来提供此类数据。在体外孵育公羊附睾（头、体、近端和远端尾）和射出（EJ）精子，以评估其经历AR的能力。光学显微镜检查显示，在穿过附睾尾近端的精子群中，≥ 50%的精子在更短的时间（< 17 h）内表现出内源性AR，而从附睾更近端区域分离的精子（22-> 50 h）则表现出内源性AR。肝素添加到精子中没有刺激附睾或EJ精子的AR，而添加钙离子载体（A23187）增加了尾部和EJ精子的AR率，但没有头部或体精子。第二个实验评估AR百分比、运动细胞百分比和仓鼠卵子穿透百分比，结果显示从附睾尾近端区域分离的精子未能穿透卵子。当尾部或EJ精子的活动率> 5%，AR > 24%时，可穿透仓鼠卵。在附睾成熟的任何阶段，精子的顶体完整性的比较，电子或光学显微镜没有不同，这表明最小的非特异性膜的变化发生，光学显微镜是有效的评估顶体状态的公羊精子。顶体酶活性（精子结合和解离）也进行了测量。总活性和释放的顶体酶从精子到培养基中在8小时的孵育过程中是大于成熟的精子。从这些实验的结果进行了讨论，必须发生在精子的变化，因为他们获得的能力，进行AR和渗透仓鼠卵。
An understanding of epididymal maturation of sperm requires descriptions of changes in membrane properties and their relation to changes in cell function. While sperm membranes have been studied in some detail in rams, few reports address associated functional changes. This report provides such data by evaluating (a) the time course of sperm acrosome reaction (AR) induction for cells from each epididymal region; (b) the capacity of epididymal sperm to penetrate ova; (c) differences in physiological AR and general sperm degeneration; and (d) acrosin release of epididymal sperm. Ram epididymal (caput, corpus, and proximal and distal cauda) and ejaculated (EJ) sperm were incubated in vitro to assess their capacity to undergo an AR. Light microscopy revealed that in sperm populations which had traversed the proximal cauda epididymidis, ≥ 50% exhibited an endogenous AR in less time (< 17 h) than did sperm isolated from more proximal regions of the epididymis (22-> 50 h). Heparin added to sperm did not stimulate the AR in epididymal or EJ sperm, whereas addition of a calcium ionophore (A23187) increased AR rates for cauda and EJ sperm, but not caput or corpus sperm. A second experiment evaluating percent AR, percent motile cells, and percent hamster ova penetrated revealed that sperm isolated from regions proximal to the cauda epididymidis failed to penetrate ova. When cauda or EJ sperm exhibited motility > 5% and AR > 24%, penetration of hamster eggs occurred. Comparisons of acrosomal integrity by electron or light microscopy were not different for sperm at any stage of epididymal maturation, suggesting that minimal nonspecific membrane changes occur and that light microscopy is valid for evaluating the acrosomal status of ram spermatozoa. Acrosin activity (sperm bound and dissociated) also was measured. Both total activity and release of acrosin from sperm to the medium during an 8-h incubation was greater for mature than for immature sperm. Results from these experiments are discussed in relation to the changes that must occur in sperm as they acquire the capacity to undergo an AR and penetrate hamster ova.