Author:
Chiarante Nicolás,Alonso Carlos A. I.,Plaza Jessica,Lottero-Leconte Raquel,Arroyo-Salvo Camila,Yaneff Agustín,Osycka-Salut Claudia E.,Davio Carlos,Miragaya Marcelo,Perez-Martinez Silvina
Abstract
AbstractPreviously we demonstrated that multidrug resistance-associated protein 4 transporter (MRP4) mediates cAMP efflux in bovine spermatozoa and that extracellular cAMP (ecAMP) triggers events associated to capacitation. Here, we deepen the study of the role of MRP4 in bovine sperm function by using MK571, an MRP4 inhibitor. The incubation of spermatozoa with MK571 during 45 min inhibited capacitation-associated events. MRP4 was localized in post-acrosomal region and mid-piece at 15 min capacitation, while at 45 min it was mainly located in the acrosome. After 15 min, MK571 decreased total sperm motility (TM), progressive motility (PM) and several kinematic parameters. The addition of ecAMP rescued MK571 effect and ecAMP alone increased the percentage of motile sperm and kinematics parameters. Since actin cytoskeleton plays essential roles in the regulation of sperm motility, we investigated if MRP4 activity might affect actin polymerization. After 15 min capacitation, an increase in F-actin was observed, which was inhibited by MK571. This effect was reverted by the addition of ecAMP. Furthermore, ecAMP alone increased F-actin levels while no F-actin was detected with ecAMP in the presence of PKA inhibitors. Our results support the importance of cAMP efflux through MRP4 in sperm capacitation and suggest its involvement in the regulation of actin polymerization and motility.
Funder
Ministry of Science, Technology and Innovation, Argentina
University of Buenos Aires
Publisher
Springer Science and Business Media LLC
Reference64 articles.
1. Chang, M. C. Fertilizing capacity of spermatozoa deposited into the fallopian tubes. Nature 168, 697–698 (1951).
2. Austin, C. R. The capacitation of the mammalian sperm. Nature 170, 326 (1952).
3. Visconti, P. E., Krapf, D., de la Vega-Beltrán, J. L., Acevedo, J. J. & Darszon, A. Ion channels, phosphorylation and mammalian sperm capacitation. Asian J. Androl. 13, 395–405 (2011).
4. Buffone, M. G., Wertheimer, E. V., Visconti, P. E. & Krapf, D. Central role of soluble adenylyl cyclase and cAMP in sperm physiology. Bioch. Biophys. Acta 1842, 2610–2620 (2014).
5. Escoffier, J. et al. Flow cytometry analysis reveals that only a subpopulation of mouse sperm undergoes hyperpolarization during capacitation. Biol. Reprod. 92, 1–11 (2015).
Cited by
5 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献