摘要
The present report describes a simple and rapid procedure to isolate highly purified sperm heads from mouse, rat and rabbit. The isolation procedure involves mincing of the caput epididymidis (CPE) of mice and rats to obtain a crude CPE suspension. This suspension is filtered to remove tubular fragments. The filtrate containing sperms and cellular contaminants is trypsinized, followed by sonication. To obtain rabbit sperm head suspension, ejaculated spermatozoa are briefly exposed to the cationic detergent CTAB-D, followed by trypsinization. Trypsin treatment of the suspension yields sperm heads and tails. The suspension containing sperm heads and tails is separated by discontinuous sucrose density gradient centrifugation. Highly purified and enriched sperm heads are recovered from the bottom of 2.5 M sucrose of the gradient. Enriched sperm heads contain less than 1% sperm tails.
摘要译文
本报告描述了一种简单且快速的程序，用于将高度纯化的精子头与小鼠，大鼠和兔分离。隔离程序涉及剁碎小鼠和大鼠的CAPUT插角（CPE），以获得粗CPE悬浮液。过滤该悬浮液以除去管状碎片。含滤液的精子和细胞污染物是胰蛋白酶化的，然后是超声处理。为了获得兔子精子头悬浮液，将射精精子短暂地暴露于阳离子洗涤剂CTAB-D，然后是胰蛋白酶化。胰蛋白酶治疗悬浮液产生弹性头和尾部。通过不连续蔗糖密度梯度离心分离含有精子头和尾部的悬浮液。从梯度的2.5米蔗糖的底部回收高度纯化和富集的精子头。富含精子头含有少于1％的精子尾。
Lee I.P. Schmid B. Zbinden G.. A Simplified Method for the Isolation of Sperm Heads from the Caput Epididymidis of Rodents and from Ejaculated Rabbit Seminal Plasma[J]. Pathobiology, 1977,45(1-2): 48-59