Departamento de Biología Celular e Histología, Facultad de Medicina, Campus de Ciencias de la Salud, Campus Mare Nostrum (CMN), Universidad de Murcia, Murcia, Spain.
Instituto Murciano de Investigación Biosanitaria Pascual Parrilla (IMIB), Campus de Ciencias de la Salud, Murcia, Spain.
Reprod Domest Anim. 2023 Aug;58(8):1164-1171. doi: 10.1111/rda.14412. Epub 2023 Jun 23.
In a previous study, our group detected the cholecystokinin (CCK) protein in the porcine oviduct. This fact, together with the involvement of CCK in the regulation of sperm protein tyrosine phosphorylation by the modulation of HCO uptake (in mice and humans) suggests a role for CCK during sperm capacitation. Therefore, on the one hand, the expression of CCK receptors (CCK1R and CCK2R) on boar testes has been investigated and probed; on the other hand, boar spermatozoa (from seminal doses of 1-day and 5-day storage) were exposed to different concentrations of CCK (0-control, 25 or 50 μM) in a medium supporting capacitation supplemented with 0, 5 or 25 mmol/L of HCO for 1 h at 38.5°C. Sperm motion (total and progressive motility), kinetic parameters, viability, acrosome status, and mitochondrial activity were determined. No differences between groups (0, 25 or 50 μM of CCK) were observed when HCO was absent in the media (p > .05). However, the results showed that when the media was supplemented with 5 mmol/L HCO in 1-day seminal dose storage, the linearity index (LIN, %), straightness index (STR, %) and oscillation index (WOB, %) (sperm kinetics parameters) increased in the presence of CCK regardless the concentration (p < .05). Nevertheless, CCK in sperm from 5-day storage only increased the WOB parameter in comparison to the control (p < .05). Furthermore, the average amplitude of the lateral displacement of the sperm head (ALH, μm) and curvilinear velocity (VCL, μm/s) decreased when CCK was present, depending on its concentration and sperm aging (1-day vs. 5-days) (p < .05). In the case of the media supporting capacitation supplemented with 25 mmol/L HCO , any differences were observed except for sperm viability in the 5-day seminal doses, which increased in the 50 μM-CCK group compared to the control (p < .05). In conclusion, these data suggest an implication of CCK protein during sperm capacitation under low bicarbonate concentration increasing the sperm linear trajectory.
在之前的一项研究中,我们小组在猪的输卵管中检测到了胆囊收缩素(CCK)蛋白。这一事实,以及 CCK 通过调节 HCO3-摄取来调节精子蛋白酪氨酸磷酸化的作用(在小鼠和人类中),表明 CCK 在精子获能过程中起作用。因此,一方面,我们研究并探测了猪睾丸上 CCK 受体(CCK1R 和 CCK2R)的表达;另一方面,我们将来自 1 天和 5 天储存的精液剂量的猪精子暴露于不同浓度的 CCK(0-对照,25 或 50 μM)中,在支持获能的培养基中补充 0、5 或 25 mmol/L 的 HCO3-,在 38.5°C 下孵育 1 小时。测定精子运动(总运动和前向运动)、动力学参数、活力、顶体状态和线粒体活性。当培养基中没有 HCO3-时(p>.05),各组(0、25 或 50 μM 的 CCK)之间没有差异。然而,结果表明,当培养基中补充 5 mmol/L HCO3-时,在 1 天储存的精液剂量中,CCK 的存在增加了线性指数(LIN,%)、直线性指数(STR,%)和摆动性指数(WOB,%)(精子动力学参数),而与浓度无关(p<.05)。然而,与对照组相比,5 天储存的精子中的 CCK 仅增加了 WOB 参数(p<.05)。此外,当存在 CCK 时,根据其浓度和精子老化(1 天与 5 天),精子头部侧向位移的平均幅度(ALH,μm)和曲线速度(VCL,μm/s)降低(p<.05)。在支持获能的培养基中补充 25 mmol/L HCO3-的情况下,除了 5 天储存的精液剂量中的精子活力增加外,任何差异都没有观察到,与对照组相比,50 μM-CCK 组的精子活力增加(p<.05)。总之,这些数据表明,在低 HCO3-浓度下,CCK 蛋白在精子获能过程中发挥作用,增加精子的线性轨迹。