Kawakami E, Vandevoort C A, Mahi-Brown C A, Overstreet J W
California Regional Primate Research Center, University of California, Davis 95616-8659.
Biol Reprod. 1993 Apr;48(4):841-5. doi: 10.1095/biolreprod48.4.841.
In this study the induction of the acrosome reaction of canine sperm by homologous zona pellucida (ZP) was examined. Twelve semen samples obtained from 6 normal beagle dogs were evaluated after sperm incubation in vitro with canine capacitation medium (CCM). Washed sperm were preincubated at 37 degrees C in 5% CO2 in air for 4 and 7 h prior to experimental treatment. Sperm were co-incubated for 1 min with intact oocytes collected from canine ovaries. Half of the oocytes were then fixed, and the bound sperm were assessed for acrosome reactions through use of a polyclonal antisperm antiserum and indirect immunofluorescence. The remaining oocytes were incubated in sperm-free medium for an additional 1-h period, and the acrosomal status of sperm bound to the ZP was evaluated similarly. The percentage of acrosome-reacted sperm on the ZP increased significantly during the 1-h incubation period. In other experiments, capacitated canine sperm were incubated with heat-solubilized ZP for 1 h and their acrosomal status was determined using fluoresceinated Pisum sativum lectin. The percentages of acrosome-reacted sperm increased significantly in ZP solution compared with controls. These data demonstrate that intact and solubilized canine ZP are capable of inducing acrosome reactions of canine sperm.
在本研究中，检测了同源透明带（ZP）对犬精子顶体反应的诱导作用。从6只正常比格犬获取12份精液样本，将精子在体外与犬获能培养基（CCM）孵育后进行评估。在实验处理前，将洗涤后的精子在37℃、5%二氧化碳的空气中预孵育4小时和7小时。精子与从犬卵巢采集的完整卵母细胞共同孵育1分钟。然后将一半的卵母细胞固定，通过使用多克隆抗精子抗血清和间接免疫荧光评估结合的精子的顶体反应。其余的卵母细胞在无精子培养基中再孵育1小时，同样评估与ZP结合的精子的顶体状态。在1小时的孵育期内，ZP上发生顶体反应的精子百分比显著增加。在其他实验中，将获能的犬精子与热溶解的ZP孵育1小时，并用荧光素标记的豌豆凝集素测定其顶体状态。与对照组相比，ZP溶液中发生顶体反应的精子百分比显著增加。这些数据表明，完整的和溶解的犬ZP能够诱导犬精子的顶体反应。
