仓鼠精子获能和运动学。

Sperm Capacitation and Kinematics in Hamsters.

机构信息

Department of Biodiversity and Evolutionary Biology, Museo Nacional de Ciencias Naturales (CSIC), Calle Jose Gutierrez Abascal 2, 28006 Madrid, Spain.

出版信息

Int J Mol Sci. 2023 Nov 8;24(22):16093. doi: 10.3390/ijms242216093.

DOI:10.3390/ijms242216093

PMID:38003282

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10671044/

Abstract

This study was designed to analyze changes in the spermatozoa of three species of hamsters incubated under different conditions. Cauda epididymal sperm were incubated for 4 h in modified Tyrode's medium containing albumin, lactate, pyruvate, and Hepes (mTALP-H), in the same medium with the addition of bicarbonate (mTALP-BH), or with bicarbonate and 20 ng/mL of progesterone (mTALP-BH+P4). Media with bicarbonate are believed to promote capacitation in rodent species. Sperm motility, viability, capacitation patterns, and kinematics were assessed at different times. Capacitation in live cells was quantified after staining with Hoechst 33258 and chlortetracycline. Patterns believed to correspond to non-capacitated cells (F pattern), capacitated, acrosome-intact cells (B pattern), and acrosome-reacted cells (AR pattern) were recognized. Kinematics were examined via computer-assisted sperm analysis (CASA). The results showed a decrease in total motility in all three species in different media, with a sharp decrease in progressive motility in bicarbonate-containing media (without or with progesterone), suggesting hyperactivated motion. However, none of the other signs of hyperactivation described in rodents (i.e., decrease in STR or LIN, together with an increase in ALH) were observed. F pattern cells diminished with time in all media and were generally lower in and higher in . B pattern cells increased in mTALP-BH media in all species. Progesterone did not enhance the percentage of B pattern cells. Finally, AR pattern cells increased in all species incubated in different media, showing the highest percentage in and the lowest in . Comparisons between media revealed that there were higher percentages of F pattern cells and lower percentages of B pattern cells over time in medium without bicarbonate (mTALP-H) in comparison to media containing bicarbonate (mTALP-BH; mTALP-BH+P4). Overall, changes consistent with the acquisition of capacitation and development of hyperactivated motility were found; however, further studies are required to better characterize media necessary to support the pathways involved in these processes in species.

摘要

本研究旨在分析在不同条件下孵育的三种仓鼠的精子发生变化。在改良的含有白蛋白、乳酸盐、丙酮酸盐和 Hepes（mTALP-H）的 Tyrode 培养基中孵育尾部附睾精子 4 小时，在含有碳酸氢盐的相同培养基中（mTALP-BH），或在含有碳酸氢盐和 20ng/mL 孕酮的培养基中（mTALP-BH+P4）。人们认为含有碳酸氢盐的培养基可以促进啮齿动物的获能。在不同时间评估精子运动性、活力、获能模式和运动学。用 Hoechst 33258 和氯四环素染色后，对活细胞中的获能进行定量。识别出与未获能细胞（F 模式）、获能、顶体完整细胞（B 模式）和顶体反应细胞（AR 模式）相对应的模式。通过计算机辅助精子分析（CASA）检查运动学。结果表明，在所有三种物种中，在不同的培养基中总运动性下降，在含有碳酸氢盐的培养基（无论是否含有孕酮）中，前向运动性急剧下降，提示超激活运动。然而，在啮齿动物中描述的其他超激活迹象（即 STR 或 LIN 减少，同时 ALH 增加）并未观察到。在所有培养基中，F 模式细胞随着时间的推移而减少，在中通常较低，在中较高。在所有物种的 mTALP-BH 培养基中，B 模式细胞增加。孕酮并未增加 B 模式细胞的百分比。最后，在所有孵育于不同培养基的物种中，AR 模式细胞增加，在中百分比最高，在中百分比最低。培养基之间的比较表明，在不含碳酸氢盐的培养基（mTALP-H）中，F 模式细胞的百分比较高，B 模式细胞的百分比较低，随着时间的推移，与含有碳酸氢盐的培养基（mTALP-BH；mTALP-BH+P4）相比。总的来说，发现了与获得获能和发展超激活运动相一致的变化；然而，需要进一步的研究来更好地描述支持这些过程相关途径所需的培养基。