Bondarenko Olga, Dzyuba Borys, Cosson Jacky, Rodina Marek, Linhart Otomar
Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, University of South Bohemia in Ceske Budejovice, Vodnany, Czech Republic,
Fish Physiol Biochem. 2014 Oct;40(5):1417-21. doi: 10.1007/s10695-014-9936-5. Epub 2014 Apr 10.
The role of environmental ion composition and osmolality in Ca(2+) signaled activation was assessed in spermatozoa of brook trout Salvelinus fontinalis. Milt from ten mature males was obtained by abdominal massage. Spermatozoa motility was evaluated in 0, 100, and 300 mOsm/kg NaCl or sucrose solutions, buffered by 10 mM Tris-HCl pH 8.5. For investigation of spermatozoa reaction to external Ca(2+) concentration, 2 mM ethylene glycol tetraacetic acid (EGTA) was added to the activation media as a calcium ions chelator. For investigation of the effect of external Na(+) concentration in conditions of low external Ca(2+), 100 µM amiloride was added to the EGTA-containing solutions as a Na(+) transport blocker. Low motility was observed in sucrose (Na(+) free) solutions containing 2 mM EGTA but not in Na(+) solutions containing 2 mM EGTA. Addition of amiloride led to significantly increased motility (P < 0.05) compared with sucrose (Na(+) free) solutions containing 2 mM EGTA. We conclude that Na(+) transport in Ca(2+)-free solutions plays a regulatory role in brook trout spermatozoa activation. The influence of competitive Na(+) and Ca(2+) transport on the control of spermatozoa activation requires further study with respect to its application for improvement of artificial activation and storage media.
在溪红点鲑(Salvelinus fontinalis)精子中评估了环境离子组成和渗透压在Ca(2+)信号激活中的作用。通过腹部按摩从10只成熟雄性获得精液。在由10 mM Tris-HCl pH 8.5缓冲的0、100和300 mOsm/kg NaCl或蔗糖溶液中评估精子活力。为了研究精子对外界Ca(2+)浓度的反应,向激活培养基中添加2 mM乙二醇四乙酸(EGTA)作为钙离子螯合剂。为了研究在低外界Ca(2+)条件下外界Na(+)浓度的影响,向含EGTA的溶液中添加100 µM氨氯吡咪作为Na(+)转运阻滞剂。在含有2 mM EGTA的蔗糖(无Na(+))溶液中观察到低活力,但在含有2 mM EGTA的Na(+)溶液中未观察到。与含有2 mM EGTA的蔗糖(无Na(+))溶液相比,添加氨氯吡咪导致活力显著增加(P < 0.05)。我们得出结论,在无Ca(2+)溶液中的Na(+)转运在溪红点鲑精子激活中起调节作用。关于竞争性Na(+)和Ca(2+)转运对精子激活控制的影响,需要进一步研究其在改善人工激活和储存培养基方面的应用。