Taniguchi Shin, Zhu Zhendong, Matsuzaki Mei, Tsudzuki Masaoki, Maeda Teruo
Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
One Health Business Department, Neopharma Japan Co., Ltd., Fujimi, Chiyodaku, Tokyo 102-0071, Japan.
Anim Biosci. 2021 Dec;34(12):1912-1920. doi: 10.5713/ab.21.0021. Epub 2021 Apr 23.
This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm.
The pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (-)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (-), PBS (-) supplemented with CaCl2 and MgCl2 (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA.
The addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period.
5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.
本研究调查了5-氨基乙酰丙酸（5-ALA）对鸡精子运动参数、线粒体膜去极化和ATP水平的影响。
使用来自 barred Plymouth Rock 雄性的混合精液。在第一个实验中，精液用含有不同浓度（0、0.01、0.05和0.1 mM）5-ALA的磷酸盐缓冲盐水（PBS（-））稀释4倍，然后通过计算机辅助精子分析（CASA）评估孵育后的精子运动参数。在第二个实验中，精液用含有0.05 mM 5-ALA的PBS（-）稀释4倍，然后分别用MitoPT® JC-1检测试剂盒和ATP检测试剂盒分析孵育1.5小时后的精子线粒体膜去极化和ATP水平。在第三个实验中，将精液从精浆中取出，并用PBS（-）、补充有CaCl2和MgCl2的PBS（+）+5-ALA、PBS（+）+咖啡因以及PBS（+）+咖啡因+5-ALA的培养基重悬。然后，通过CASA评估孵育后的精子运动参数。在最后一个实验中，在去除精浆后，精液用PBS（-）、PBS（-）+5-ALA、5.7%葡萄糖、5.7%葡萄糖+5-ALA的培养基处理，然后通过CASA评估精子运动参数。
添加0.05 mM 5-ALA显著提高了鸡精子的运动能力、渐进运动能力、线性度、平均路径速度、曲线速度、直线速度和摆动。5-ALA处理也增加了精子线粒体膜去极化。5-ALA处理降低了精子ATP水平。咖啡因处理和葡萄糖处理均降低了孵育期间的精子运动能力。
5-ALA可能增加精子线粒体膜去极化，以利用ATP增强精子运动。
