Romarowski Ana, Sánchez-Cárdenas Claudia, Ramírez-Gómez Héctor V, Puga Molina Lis del C, Treviño Claudia L, Hernández-Cruz Arturo, Darszon Alberto, Buffone Mariano G
Instituto de Biología y Medicina Experimental (IBYME), National Research Council of Argentina (CONICET), Buenos Aires, Argentina.
Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México (UNAM), Cuernavaca, Morelos, México.
Biol Reprod. 2016 Mar;94(3):63. doi: 10.1095/biolreprod.115.136085. Epub 2016 Jan 27.
During capacitation, sperm acquire the ability to undergo the acrosome reaction (AR), an essential step in fertilization. Progesterone produced by cumulus cells has been associated with various physiological processes in sperm, including stimulation of AR. An increase in intracellular Ca(2+) ([Ca(2+)]i) is necessary for AR to occur. In this study, we investigated the spatiotemporal correlation between the changes in [Ca(2+)]i and AR in single mouse spermatozoa in response to progesterone. We found that progesterone stimulates an [Ca(2+)]i increase in five different patterns: gradual increase, oscillatory, late transitory, immediate transitory, and sustained. We also observed that the [Ca(2+)]i increase promoted by progesterone starts at either the flagellum or the head. We validated the use of FM4-64 as an indicator for the occurrence of the AR by simultaneously detecting its fluorescence increase and the loss of EGFP in transgenic EGFPAcr sperm. For the first time, we have simultaneously visualized the rise in [Ca(2+)]i and the process of exocytosis in response to progesterone and found that only a specific transitory increase in [Ca(2+)]i originating in the sperm head promotes the initiation of AR.
在获能过程中,精子获得了发生顶体反应(AR)的能力,这是受精过程中的关键步骤。卵丘细胞产生的孕酮与精子的多种生理过程相关,包括刺激顶体反应。细胞内钙离子浓度([Ca(2+)]i)升高是顶体反应发生所必需的。在本研究中,我们调查了单个小鼠精子中[Ca(2+)]i变化与孕酮诱导的顶体反应之间的时空相关性。我们发现孕酮以五种不同模式刺激[Ca(2+)]i升高:逐渐升高、振荡、延迟短暂升高、立即短暂升高和持续升高。我们还观察到,孕酮诱导的[Ca(2+)]i升高始于鞭毛或头部。通过在转基因EGFP-Acr精子中同时检测FM4-64荧光增加和EGFP丢失,我们验证了FM4-64可作为顶体反应发生的指标。首次,我们同时观察到了[Ca(2+)]i升高和孕酮诱导的胞吐过程,发现只有源于精子头部的特定短暂[Ca(2+)]i升高促进顶体反应的起始。