PDF(Pubmed)
Abstract：
ProAKAP4, a precursor of AKAP4 (A-kinase anchor protein) found in the flagellum of mammalian and non-mammalian spermatozoa, serves as a structural protein with established correlations to motility parameters across diverse species. This study aimed to determine the proAKAP4 level evolution in thawed stallion semen over a 3 h period, examining its correlation with motility descriptors and mitochondrial membrane potential. Utilizing sixteen ejaculates from four French warmblood stallions, this study involved maintaining thawed samples at 37 °C for 3 h, conducting proAKAP4 enzyme-linked immunosorbent assays (ELISA), computer-assisted sperm analysis (CASA), and mitochondrial membrane potential by JC-1 probe and flow cytometry at 0, 1, and 3 h post-thawing. The findings indicate significant positive correlations (p ≤ 0.05) between proAKAP4 levels and sperm total or progressive motility at all time points analyzed. Spermatozoa velocity descriptors (VAP, VCL, VSL) and spermatozoa lateral head displacement (ALH) display positive correlations (p ≤ 0.05) with ProAKAP4 at the 0 h post-thawing. ProAKAP4 concentration exhibits no discernible difference between batches with or without a cryoprotectant. Notably, proAKAP4 consumption remains insignificant within the initial hour after thawing but becomes significant (p ≤ 0.05) between 1 and 3 h post-thawing. In summary, proAKAP4 demonstrates positive correlations with total and progressive motility in stallion semen for up to 3 h after thawing, albeit showing a noticeable decrease starting from the first hour post-thawing, indicating a progressive consumption as a result of spermatozoa motile activity.
摘要：
ProAKAP4，在哺乳动物和非哺乳动物精子的鞭毛中发现的AKAP4（A激酶锚定蛋白）的前体，作为一种结构蛋白，与不同物种的运动性参数建立了相关性。本研究旨在确定3小时内解冻的种马精液中的proAKAP4水平进化。检查其与运动性描述符和线粒体膜电位的相关性。利用四个法国温血种马的16次射精，这项研究涉及将解冻的样品在37°C下保持3小时，进行proAKAP4酶联免疫吸附测定（ELISA），计算机辅助精子分析（CASA），解冻后0、1和3小时，通过JC-1探针和流式细胞术检测线粒体膜电位。研究结果表明，在所有分析的时间点，proAKAP4水平与精子总运动性或进行性运动性之间存在显着正相关（p≤0.05）。精子速度描述符(VAP，VCL,VSL）和精子侧头位移（ALH）在解冻后0h与ProAKAP4呈正相关（p≤0.05）。ProAKAP4浓度在具有或不具有冷冻保护剂的批次之间没有表现出可辨别的差异。值得注意的是,proAKAP4的消耗在解冻后的最初小时内仍然微不足道，但在解冻后1至3小时之间变得显着（p≤0.05）。总之,proAKAP4在解冻后3小时内与种马精液的总运动和进行性运动呈正相关，尽管从解冻后的第一个小时开始显示出明显的下降，表明由于精子活动而逐渐消耗。
