Molecular and Cellular Endocrinology 187 (2002) 139 – 144 Intracellular calcium regulation in sperm capacitation and acrosomal reaction Haim Breitbart * Faculty of Life Sciences, Bar -Ilan Uniersity, Ramat Gan 52900, Israel Abstract Binding to the egg’s zona pellucida stimulates the spermatozoon to undergo acrosome reaction, a process which enables the sperm to penetrate the egg. Prior to this binding, the spermatozoa undergo in the female reproductive tract a series of biochemical transformations, collectively called capacitation. The first event in capacitation is the elevation of intracellular calcium and bicarbonate to activate adenylyl cyclase (AC) to produce cyclic-AMP, which activates protein kinase A (PKA) to phosphorylate certain proteins. During capacitation, there is also an increase in actin polymerization and in the membrane-bound phospholipase C (PLC). Sperm binding to zona-pellucida causes further activation of cAMP/PKA and protein kinase C (PKC), respectively. PKC opens a calcium channel in the plasma membrane. PKA together with inositol-trisphosphate activate calcium channels in the outer acrosomal membrane, which leads to an increase in cytosolic calcium. The depletion of calcium in the acrosome will activate a store-operated calcium entry mechanism in the plasma membrane, leading to a higher increase in cytosolic calcium, resulting in membrane fusion and acrosome reaction. © 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Calcium; Regulation; Sperm; Capacitation; Acrosome reaction www.elsevier.com/locate/mce 1. Introduction Mammalian spermatozoa are unable to fertilize the egg immediately after ejaculation. They require a period of incubation in the female reproductive tract in order to acquire the capacity to fertilize. During this time, the spermatozoa undergo a poorly defined process of matu- ration known as capacitation. There is no clear recog- nizable marker for the occurrence of capacitation, although several intracellular changes are known to occur including increases in membrane fluidity, choles- terol efflux, intracellular Ca 2 + and cAMP concentra- tions, protein tyrosine phosphorylation, and changes in swimming patterns and chemotactic motility. Binding to the zona pellucida stimulates the sperma- tozoon to undergo the acrosome reaction in which the outer acrosomal membrane fuses with the overlying plasma membrane (Yanagimachi, 1995). This exocy- totic event results in the release of hydrolytic enzymes, principally, the trypsin like acrosin, and in the exposure of new membrane domains, both of which are essential for the fertilization process. In order to penetrate the zona pellucida the spermatozoon creates a groove which is approximately the width and height of the head (Allen and Green, 1997). Here an overview of the processes involved in intracellular Ca 2 + regulation re- sponsible for the acrosome reaction is provided. 2. Protein tyrosine kinase Tyrosine phosphorylation of proteins plays an im- portant role in capacitation (Visconti and Kopf, 1998), acrosome reaction and sperm penetration (Burks et al., 1995). Inhibitors of protein kinase A (PKA) inhibit both protein tyrosine phosphorylation and capacitation of spermatozoa (Visconti et al., 1995). In human sper- matozoon, reactive oxygen species (ROS) up-regulate protein tyrosine phosphorylation of several proteins (Aitken et al., 1995; Leclerc et al., 1997). These obser- vations are consistent with the suggestion that H 2 O 2 activates adenylyl cyclase to produce cAMP and the subsequent activation of the serine/threonine kinase A (Aitken, 1997). It has been suggested that Ca 2 + and HCO 3 - are also required for the activation of sperm adenylyl cyclase (Visconti et al., 1995). * Tel.: +972-3-5318201; fax: +972-3-5344766. E-mail address: breith@mail.biu.ac.il (H. Breitbart). 0303-7207/02/$ - see front matter © 2002 Elsevier Science Ireland Ltd. All rights reserved. PII:S0303-7207(01)00704-3