Mendoza Francisco J, Perez-Marin Carlos C, Garcia-Marin Luis, Madueño Juan A, Henley Charles, Aguilera-Tejero Escolastico, Rodriguez Mariano
Dept Medicina y Cirugia Animal, Universidad de Cordoba, Campus Universitario Rabanales, Ctra. Madrid-Cadiz km 396, Cordoba, Spain.
J Androl. 2012 Jan-Feb;33(1):96-104. doi: 10.2164/jandrol.110.011254. Epub 2011 Mar 10.
The intracellular movement of calcium, through calcium channels, plays a major role on sperm cell function. The calcium-sensing receptor (CaSR), a molecular mechanism by which many cells detect changes in extracellular calcium concentration, has not been described in spermatozoa. The aim of this study was to investigate the expression of the CaSR in testicular tissue and sperm cells and the functional consequences of spermatozoid CaSR activation by calcimimetics. CaSR mRNA and protein were identified both in rat testicular tissue and in rat spermatozoa using real-time reverse transcription-polymerase chain reaction and immunohistochemistry, respectively. Functionality of CaSR was evaluated by studying the influence of calcimimetic AMG 641 on rat and pig sperm motility. Treatment with AMG 641 100 nM for 1 hour increased rat sperm motility from a score of 1.0 ± 0.1 to 3.8 ± 0.3 (P < .05). AMG 641 also resulted in a modest but significant increase in the pig sperm motility parameters evaluated by computer-assisted sperm analysis. AMG 641 was effective in a wide range of concentrations but resulted in a more marked effect at 50-100 nM. In addition, AMG 641 did not have any negative effect on sperm viability, which was measured by flow cytometry. In conclusion, our results demonstrate the expression of functional CaSR in testicular tissue and sperm, which can be activated by calcimimetic AMG 641.
钙通过钙通道进行的细胞内移动在精子细胞功能中起主要作用。钙敏感受体（CaSR）是许多细胞检测细胞外钙浓度变化的一种分子机制，在精子中尚未见报道。本研究的目的是调查CaSR在睾丸组织和精子细胞中的表达情况，以及拟钙剂激活精子CaSR的功能后果。分别使用实时逆转录-聚合酶链反应和免疫组织化学方法，在大鼠睾丸组织和大鼠精子中鉴定出了CaSR mRNA和蛋白。通过研究拟钙剂AMG 641对大鼠和猪精子活力的影响来评估CaSR的功能。用100 nM的AMG 641处理1小时，大鼠精子活力评分从1.0±0.1提高到3.8±0.3（P<.05）。通过计算机辅助精子分析评估，AMG 641也使猪精子活力参数有适度但显著的增加。AMG 641在很宽的浓度范围内都有效，但在50-100 nM时效果更明显。此外，通过流式细胞术检测，AMG 641对精子活力没有任何负面影响。总之，我们的结果表明功能性CaSR在睾丸组织和精子中表达，且可被拟钙剂AMG 641激活。
