Holden C A, Trounson A O
Centre for Early Human Development, Monash Medical Centre, Clayton, Victoria, Australia.
J Androl. 1992 May-Jun;13(3):260-5.
To achieve higher rates of acrosomal loss for in vitro studies of human sperm function, the effect of liposomes prepared from the phospholipid, dilauroylphosphatidylcholine (PC12), on human spermatozoa was investigated. In general, acrosome loss was induced with PC12 with an associated decline in the percentage of motile spermatozoa. Reduction of bovine serum albumin concentration in the incubation medium from 12 mg/mL to 3 mg/mL resulted in a more pronounced effect of PC12, with a significant reduction (P less than 0.001) in the percentage of motile spermatozoa within 1 hour of incubation with PC12. The percentage of spermatozoa observed to be acrosome-free under staining with fluorescein-conjugated Concanavalin A lectin was significantly reduced (P less than 0.05) when spermatozoa were incubated with PC12 (260 mumol/L) in the absence of calcium. The percentage of motile spermatozoa was not different during PC12 incubations when Ca2+ concentration (0, 2.5, and 5 mmol/L) was altered. An active motility pattern was restored in PC12-treated human spermatozoa by subsequent incubation in a defined medium containing 7.5 mmol/L adenosine 5'-triphosphate and 20 mumol/L cyclic adenosine 3', 5'-monophosphate. It was demonstrated that PC12-treated human spermatozoa were capable of binding to the zona pellucida of salt-stored human oocytes once an active motility pattern had been restored.
为了在人类精子功能的体外研究中实现更高的顶体丢失率，研究了由磷脂二月桂酰磷脂酰胆碱（PC12）制备的脂质体对人类精子的影响。一般来说，PC12可诱导顶体丢失，同时活动精子的百分比会下降。将孵育培养基中牛血清白蛋白的浓度从12mg/mL降至3mg/mL，会使PC12的作用更显著，在与PC12孵育1小时内，活动精子的百分比显著降低（P小于0.001）。当精子在无钙条件下与PC12（260μmol/L）孵育时，用荧光素偶联的刀豆球蛋白A凝集素染色观察到的无顶体精子百分比显著降低（P小于0.05）。当改变Ca2+浓度（0、2.5和5mmol/L）时，PC12孵育期间活动精子的百分比没有差异。通过随后在含有7.5mmol/L腺苷5'-三磷酸和20μmol/L环腺苷3'，5'-单磷酸的特定培养基中孵育，可使经PC12处理的人类精子恢复活跃的运动模式。结果表明，一旦恢复活跃的运动模式，经PC12处理的人类精子能够与盐储存的人类卵母细胞的透明带结合。
