精子动力蛋白atp酶和与各种细胞活动相关的atp酶(AAA+):作为极端精子运动障碍的少弱畸形精子症和失活精子症的调节

S. W. Lestari, F. Firdaus, Dessy Noor Miati, Asmarinah
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摘要

弱精子症是最常见的精子运动障碍,但也有其他更极端的精子运动障碍,即少弱畸形精子症(OAT)和无精子症。有几种已知的OAT和无精症的细胞机制,但关于动力蛋白atp酶和atp酶与各种细胞活动相关的数据有限(AAA+)。AAA1参与ATP水解,AAA2在ATP结合口袋中纠缠。本研究旨在探讨动力蛋白atp酶活性的作用及AAA动力蛋白的定量测定。本研究使用了14名OAT患者、11名失活精子患者和17名无活精子患者的精子样本。采用Makler腔测定精子浓度和活力,采用Papanicolaou染色精液涂片,采用世界卫生组织第五版标准测定精子形态,通过计算释放的无机磷酸盐定量测定动力蛋白atp酶。采用酶联免疫吸附法定量,免疫细胞化学法测定其分布。结果表明,OAT组和无精子症组的动力蛋白atp酶活性(分别为2.68±0.76、1.01±0.31、7.22±1.08 μmol Pi/mg protein/h)低于正常精子症组(P<0.05), AAA1和AAA2含量也低于正常精子症组(P<0.05)。此外,精子尾部AAA的染色与动力蛋白atp酶的活性和数量相一致,在正常精子样本中AAA的含量最高,在OAT样本中含量较低,在坏死性精子样本中几乎检测不到。受损精子动力蛋白的结构和功能可能改变动力蛋白atp酶活性和AAA1和AAA2水平。
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Sperm dynein ATPase and ATPases associated with various cellular activities (AAA+): regulation in oligo-astheno-teratozoospermia and necrozoospermia as extreme sperm motility disorders
Asthenozoospermia is the most frequent sperm motility disorder, but there are other more extreme sperm motility disorders, namely oligo-astheno-teratozoospermia (OAT) and necrozoospermia. There are several cellular mechanisms known for OAT and necrozoospermia, but there are limited data on dynein ATPase and ATPases associated with various cellular activities (AAA+). AAA1 is involved in ATP hydrolysis, while AAA2 is entangled in ATP-binding pocket. This study was conducted to investigate the role of dynein ATPase activity and quantification of AAA dynein. Spermatozoa from 14 men with OAT, 11 men with necrozoospermic and 17 men with normoozspermic samples were used in this study. Makler chamber was used to determine sperm concentration and motility, while Papanicolaou stained semen smears using World Health Organization-fifth edition criteria was performed to determine sperm morphology, and dynein ATPase was quantified by calculation of released inorganic phosphate. AAA was quantified by enzyme-linked immunosorbent assay, whereas the distribution was determined by immunocytochemistry. This study showed that the dynein ATPase activity in OAT and necrozoospermia was lower than in the normozoospermic group (2.68±0.76, 1.01±0.31, 7.22±1.08 μmol Pi/mg protein/h, respectively, P<0.05), as well as the amounts of AAA1 and AAA2. In addition, staining for AAA in the sperm tail paralleled the dynein ATPase activity and quantity of AAA, being the highest in sperm from normozoospermic samples, lower in sperm from OAT samples, and almost undetectable in sperm from necrozoospermic samples. The structure and function of damaged sperm dynein may alter dynein ATPase activity and levels of AAA1 and AAA2.
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