Manuscript accepted on :February 10, 2018
Published online on:
Sperm Na+ K+-ATPase and Ca2+-ATPase Activities: A Potential Predictive Parameter of Sperm Motility Disorder in Infertile Men
Silvia W Lestari1,4, Manggiasih D. Larasati2, Indra G. Mansur1,4 and Ria Margiana3,4
1Department of Medical Biology, Faculty of Medicine Universitas Indonesia.
2Master Program in Biomedical Sciences, Faculty of Medicine Universitas Indonesia.
3Department of Anatomy, Faculty of Medicine Universitas Indonesia.
4The Indonesian Reproductive Medicine Research and Training Center, Ina-Repromed.
Corresponding Author E-mail: email@example.com
Current highlight on the etiology of male infertility is a disorder in sperm motility which is caused by ion homeostasis imbalance involving the ubiquitous multifunctional transmembrane protein, Na+K+-ATPase and Ca2+-ATPase enzymes. The emphasis of this review is evaluating the use of sperm’s Na+K+-ATPase and Ca2+ATPase activity as predictive parameters of sperm motility disorder. To this purpose, a computerized search of PubMed database was performed and obtained data were reviewed in this paper. The retrieved studies were laboratory experiments involving human and mice sperm as the subjects. Na+ K+-ATPase and Ca2+-ATPase play an essential role in sperm motility by controlling ion homeostasis. Na+ K+-ATPase maintains the intracellular pH by transporting 3 Na+ out and 2 K+ into the cell, whereas Ca2+-ATPase extrudes Ca2+ from the cell. The impairment of these enzymes and its isoforms, Na+ K+-ATPase α4 and PMCA4 expression were proved to decrease sperm motility.
Ca2+ATPase Male Infertility; Na+ K+-ATPase; Sperm Motility;Download this article as:
|Copy the following to cite this article:
Lestari S. W, Larasati M. D, Mansur I. G, Margiana R. Sperm Na+ K+-ATPase and Ca2+-ATPase Activities: A Potential Predictive Parameter of Sperm Motility Disorder in Infertile Men. Biomed Pharmacol J 2018;11(1).
|Copy the following to cite this URL:
Lestari S. W, Larasati M. D, Mansur I. G, Margiana R. Sperm Na+ K+-ATPase and Ca2+-ATPase Activities: A Potential Predictive Parameter of Sperm Motility Disorder in Infertile Men. Biomed Pharmacol J 2018;11(1). Available from: http://biomedpharmajournal.org/?p=19063
Male infertility is an inability of an individual male to make his partner conceive which is caused by the presence of an abnormal semen analysis parameters after one year having regular, no-contraceptives-used intercourse. For a success in obtaining pregnancy, there are several things that need to occur. At first, the male has to produce a healthy sperm which depends on how the reproductive organ grow and form during puberty. At least one of the testicles must function properly for achieving pregnancy. Moreover, endocrine homeostasis such as androgen production has to be maintained in order to trigger sperm production.
The sperm must be carried out into the semen in a number sufficient enough to function properly in achieving fertility. In case the number of sperm in the semen is less than normal, it may reduce the chances of the sperm fertilizing the partner’s oocyte. Lastly, the sperm must be functional and have a proper motility. In the case of abnormal sperm motility termed asthenozoospermia, the sperm may fail to penetrate or even reach the partner’s oocyte. The etiology of this disorder involves an array of biochemical and functional defects.1
The complexity of sperm motility involves the mounting of proteins and ions in appropriate concentration which are critical for molecular mechanisms, thus generating the sperm flagella movement.2 Among them, Na+ K+-ATPase and Ca2+ATPase are an enzyme which are essential in maintaining the membrane gradient and conducting sperm motility such as activation and hyperactivation.
The Na+ K+-ATPase is the transmembrane protein that is responsible for controlling ion homeostasis involving an active transport of Na and K ions across the plasma membranes of most cells, including sperm.5 Another ion, calcium, an important cellular second messenger, is essential for sperm motility which is initially controlled by the activity of the plasma membrane Ca2+-ATPase (PMCA), by extruding Ca2+ from the cells against the membrane gradient. Disruption of any or both of this process, Na+ K+-ATPase, and Ca2+-ATPase homeostasis activities, may contribute to sperm motility failure and probably driving to infertility in male.7,8
Sperm Na+ K+-ATPase and Ca2+-ATPase as a Potential Predictive Parameter of Sperm Motility Disorder
Na+ K+-ATPase Role in Sperm Motility
Na+ K+-ATPase’s role as an ion transporter in the sperm involves the active exchange of intracellular Na+ for extracellular K+ in sperm in order to generate Na+ and K+ gradients which is essential for maintaining the cellular ion homeostasis, including the cell membrane resting potential at -70mV, cell volume, and the transport of solutes passing through the cell membrane. Na+ K+-ATPase is constitutes of a heterodimer of two catalytic molecular variants, termed α and β subunits, which is involved in the ATP hydrolysis of Na+ K+-ATPase.1
Data reported that four α and three β subunits were identified as an isoform of Na+ K+-ATPase in mammals.10 The subunit α polypeptide has been extensively investigated regarding its role in the ATP hydrolysis and ion-translocation functions of Na+ K+-ATPase. Kocak-Toker et al,4 have provided evidence for the existence of α4 along with α1 isoform which are expressed in the mid-piece of the sperm flagella. By forming active complexes consisting of α4 subunit along with β1 and β3 subunits, the catalytic activity of Na+ K+-ATPase is being carried out.
Numerous researches had investigated the role and correlation between Na+ K+-ATPase and sperm motility as presented in Table 1. (Table 1) The experimental researches were carried out by using human and mouse sperm. Jimenez et al1 reported that the inhibition of Na+ K+-ATPase α4 isoform by ouabain was sufficient to decrease the motility of sperm significantly. Other researches have strengthened the findings that there was a significant relation between Na+ K+-ATPase activity and sperm motility performance.12,13 The findings in unison agreed that Na+ K+-ATPase has a crucial role in the functioning of a properly-working sperm.
Table 1: Role of Na+ K+-ATPase in sperm motility
|Na+ K+-ATPase role in sperm motility||Reference|
|Ion homeostasis is controlled by the activity of Na+ K+-ATPase defining the sperm motility.||Jimenez T, et al., 2011 |
|Inhibition of Na+ K+ activity poses a significant reduction in sperm motility||Peralta-Arias, et al., 2015 |
|Ouabain incubation of mouse and human sperm performed selective inhibition to α4 and sufficient to significantly decrease sperm motility||Yang YH, et al., 2014 15
Jimenez T, et al., 2012 
|Na+ K+-ATPase α4 isoform plays a crucial role in sperm motility.||Camara, et al., 2017 |
Table 2: Mechanism of Na+ K+-ATPase in sperm motility
|Na+ K+-ATPase mechanism in sperm motility||Reference|
|Catalyzing the exchange of cytoplasmic Na+ for extracellular K+ in 3:2 ratio.||Suhail, 2010 |
|NHE exchanges H+ out of the cell for extracellular Na+ while also regulating intracellular pH.||Martins AD, et al., 2014 |
|Shift in the intracellular pH to an acidic state reduces sperm motility||Ji Zhou, et al., 2015 
Zhang, et al., 2017
The mechanism of Na+ K+-ATPase in sperm motility varies, as demonstrated in Table 2. (Table 2) The Na+ K+-ATPase has a role in the active transport of Na+ and K+ across the plasma membranes by catalyzing the exchanging movement of cytoplasmic Na+ and extracellular K+ in 3:2 ratio.17 Furthermore, another research conducted by Martins et al,19 proved that Na+ H+ exchangers regulated the exchange of intracellular H+ out of the cell and extracellular Na+ into the cell in order to regulate intracellular pH, abolishing excess acid from the cells. Zhang et al,23 proposed that H+ efflux was related to Na+ influx, as performed during the initiation of motility where H+ were extruded from sperm. The alteration of intracellular pH to acidic state leads to sperm motility reduction.
The authors assessed that the proper functioning of the sperm relies on the regulation of Na+ K+-ATPase activity and its isoforms in order to support and maintain membrane potential, motility changes, and hyperactivation during capacitation, all of which played a crucial role in male fertility. In addition, based on previously researches, there is a possibility to utilize Na+ K+-ATPase α4 isoform as a potential biomarker for male infertility, and later also considered as a promising agent of male contraception.
Ca2+-ATPase Role in Sperm Motility
In achieving of successful fertilization, the sperm have to pass through reproductive tract and undergoes capacitation prior to the penetration of the oocyte. Afterward, the sperm would interact with the zona pellucida and release acrosome material. Calcium is suggested as an important ion to exert the normal sperm function in all of the process. Calcium as one of important second messenger is essential in various sperm functions, one of which is associated with sperm motility, particularly activation and hyper-activation. In sperm, calcium ion homeostasis is under strict control of the calcium pump, located in the head and tail of the sperm, which is due to the activity of the plasma membrane Ca2+-ATPase (PMCA).2
Numerous researches investigated the role of Ca2+-ATPase in sperm motility as presented in Table 3. (Table 3) Vignini et al,24 investigated that calcium ion homeostasis in physiological cell function which is the responsibility of the calcium pump, is performed by Ca2+-ATPase, which is an ATPase that has a role in extrusion Ca2+ out of the cell.26 PMCA4 was found in the principal piece of flagellum and has a certain errand in sperm motility and hyperactivity.28 The inhibition by quercetin, as an inhibitor of PMCA, leads to a reduction of sperm motility.30-32 This inhibition probably caused infertility in men.28 Another finding showed that incubation of sperm using cadmium, a compound which is found in seminal plasma of smoker, is associated with significant reduction of sperm motility afflicting Ca2+-ATPase and axonemal dynein-ATPase.2
Table 3: Role of Ca2+-ATPase in sperm motility
|Ca2+-ATPase role in sperm motility||Reference|
|Ca2+-ATPase responsible for maintaining calcium ion homeostasis in the cell||Vignini, et al., 2009 |
|The plasma membrane Ca2+ pump (PMCA) was an ATPase that extrudes Ca2+ out of the cell||Brini M and Carafoli E, 2011 |
|Quercetin incubation of human sperm could reduce sperm motility||Liang X, et al., 2016 |
|Cadmium incubation diminishes Ca2+-ATPase activity of human sperm and reduce sperm motility||Da Costa et al, 2015 |
|PMCA4 had a certain errand in sperm motility and hyperactivity and PMCA4 dysfunction caused infertility in men.||Patel R, et al., 2013 
Andrews RE, et al., 2015 
Table 4: Mechanism of Ca2+-ATPase in sperm motility
|Ca2+-ATPase mechanism in sperm motility||Reference|
|Activation of Ca2+/H+ transport by PMCA generating Ca2+ extrusion into the mitochondrial matrix and resulting in pH decrease.||Poburko, et al., 2011 |
|Calcium ion homeostasis was linking to cell metabolism by mitochondria.||Boczek, et al., 2014 |
|The physiological and pathological calcium signals were modulated by the activity of mitochondria||Chalmers, et al., 2008 ; McKenzie, et al., 2004 |
|Mitochondria altered the promulgation of calcium ion transport, and enable the restocking of intracellular calcium ion stores.||Malli R, et al., 2003 |
The mechanism of Ca2+-ATPase in sperm motility is demonstrated in Table 4. Poburko et al,34 described that PMCA activity as a Ca2+/H+ counter transport with a 1:1 stoichiometric proportion produced a large amount of calcium protons extrusion into the mitochondrial matrix and resulting in pH decrease. Whereas, Boczek et al,35 explained that calcium ion homeostasis was also linked to cell metabolism by mitochondria. The physiological and pathological calcium signals were modulated by the activity of mitochondria, i.e. by buffering the intracellular calcium ions and by synchronizing the Ca2+-dependent activation or inhibition of numerous processes Chalmers et al,36 By acting as a transient calcium ion buffers, the mitochondria will alter the promulgation of calcium ion transport, modify the activity of plasma membrane channels and transporters and enable the restocking of intracellular calcium ion stores.37,38
The authors concluded that Ca2+-ATPase is involved in controlling the intracellular calcium concentration as well as a major parameter which take part in the mechanism regulating sperm motility. Ca2+-ATPase could be used as a potential biomarker for analyzing either genetic or environmental causes of male infertility. In addition, exposure of inhibitory agents against Ca2+-ATPase proved in decreasing sperm motility and further leading to infertility. In the future, the development of inhibitory agents for targeting the Ca2+-ATPase by mimicking the effect of gene deletion on sperm motility may act as contraceptive drugs, yet the further research is required.
In conclusion, Na+, K+-ATPase and Ca2+-ATPase have a role in ion homeostasis which is required in the physiological function of cell including sperm especially for sperm motility. Furthermore, Na+ K+-ATPase has a potential as a biomarker of male infertility, thus could also be considered as a promising agent of male contraception. On the other hand, Ca2+-ATPase as calcium pump which is responsible for calcium homeostasis has a role in initiation of motility leading to acrosome reaction. The impairment of Na+ K+-ATPase and Ca2+-ATPase activities and the expression of Na+ K+-ATPase α4 and PMCA4 may be considered as predictive parameters of sperm motility disorder.
The authors are grateful to Hibah PITTA DRPM 2017 for supporting this study and research assistant Meidika Dara Rizki, Debby Aditya and Muhammad Iqbal Adi Pratama for editorial assistance.
Conflict of Interest
There is no conflict of interest.
Funding support for this study was received from Hibah Publikasi Terindeks Internasional untuk Tugas Akhir Mahasiswa (PITTA), Direktorat Riset dan Pengabdian Masyarakat (DRPM) Universitas Indonesia, 2017.
- Jimenez T, McDermott J.P, Sánchez G, Blanco G. Na, K-ATPase α4 isoform is essential for sperm fertility. Proc Natl Acad Sci U S A. 2011;108(2):644-9.
- Costa D.R, Botana D, Pinero S, Proverbio F, Marín R. Cadmium inhibits motility, activities of plasma membrane Ca2+‐ATPase and axonemal dynein‐ATPase of human spermatozoa. Andrologia. 2016;48(4):464-9.
- Kaplan J.H. Biochemistry of Na, K-ATPase. Annu Rev Biochem. 2002;71(1):511-35.
- Koçak‐toker N, Aktan G, Aykaç‐toker G. The role of Na, K‐ATPase in human sperm motility. Int .J. Androl. 2002;25(3):180-5.
- Thundathil J.C, Rajamanickam G.D, Kastelic J.P, Newton L.D. The effects of increased testicular temperature on testis-specific isoform of Na+/K+ -ATPase in sperm and its role in spermatogenesis and sperm function. Reprod Domest Anim. 2012;47(4):170-7.
- Woo A.L, James P.F, Lingrel J.B. Sperm motility is dependent on a unique isoform of the Na, K-ATPase. J Biol Chem. 2000;275(27):20693-9.
- Andrews R.E, Galileo D.S, Martin-DeLeon P.A. Plasma membrane Ca2+-ATPase 4: interaction with constitutive nitric oxide synthases in human sperm and prostasomes which carry Ca2+/CaM-dependent serine kinase. Mol Hum Reprod. 2015;21(11):832-43.
- McDermott J, Sanchez G, Nangia A.K, Blanco G. Role of human Na,K-ATPase alpha 4 in sperm function, derived from studies in transgenic mice. Mol Reprod Dev. 2015;82(3):167-81.
- Jorgensen P.L, Håkansson K.O, Karlish S.J. Structure and mechanism of Na, K-ATPase: functional sites and their interactions. Annu Rev Physiol. 2003;65(1):817-49.
- Geering K. Functional roles of Na, K-ATPase subunits. Curr Opin Nephrol Hypertens. 2008;17(5):526-32.
- Sanchez G, Nguyen A.N.T, Timmerberg B, Tash J.S, Blanco G. The Na, K-ATPase α4 isoform from humans has distinct enzymatic properties and is important for sperm motility. Mol Hum Reprod. 2006;12(9):565-76.
- Peralta-Arias R.D, Vivenes C.Y, Camejo M.I, Pinero S, Proverbio T, Martinez E, et al. ATPases, ion exchangers and human sperm motility. Reproduction. 2015;149(5):475-84.
- Camara D.R, Kastelic J.P, Thundathil J.C. Role of the Na(+)/K(+)-ATPase ion pump in male reproduction and embryo development. Reprod Fertil Dev. 2017;29(8):1457-67.
- Yang Y.H, Wan Y, Lou H, Xue T, Su P. Relationship between ouabain and asthenozoospermia. J. Huazhong Univ Sci Technolog Med Sci. 2014;34(1):87-90.
- Jimenez T, Sánchez G, Wertheimer E, Blanco G. Activity of the Na, K-ATPase α4 isoform is important for membrane potential, intracellular Ca2+, and pH to maintain motility in rat spermatozoa. Reproduction. 2010;139(5):835-45.
- Jimenez T, Sánchez G, Blanco G. Activity of the Na, K‐ATPase α4 Isoform Is Regulated During Sperm Capacitation to Support Sperm Motility. J. Androl. 2012;33(5):1047-57.
- Suhail M. Na+, K+-ATPase: ubiquitous multifunctional transmembrane protein and its relevance to various pathophysiological conditions. J. Clin Med Res. 2010;2(1):1.
- Hamamah S, Gatti J-L. Role of the ionic environment and internal pH on sperm activity. Hum Reprod. 1998;13(4):20-30.
- Martins A.D, Bernardino R.L, Neuhaus-Oliveira A, Sousa M, Sá R, Alves M.G, et al. Physiology of na+/h+ exchangers in the male reproductive tract: relevance for male fertility. Biol Reprod. 2014;91(1):11,1-6.
- Wong P, Lee W, Tsang A. The effects of extracellular sodium on acid release and motility initiation in rat caudal epididymal spermatozoa in vitro. Exp Cell Res. 1981;131(1):97-104.
- Giroux‐Widemann V, Jouannet P, Pignot‐Paintrand I, Feneux D. Effects of pH on the reactivation of human spermatozoa demembranated with triton X‐100. Mol Reprod Dev. 1991;29(2):157-62.
- Zhou J, Chen L, Li J, Li H, Hong Z, Xie M, et al. The semen pH affects sperm motility and capacitation. PLoS One. 2015;10(7):e0132974.
- Zhang Z, Yang Y, Wu H, Zhang H, Zhang H, Mao J, et al. Sodium-Hydrogen-Exchanger expression in human sperm and its relationship with semen parameters. J. Assist Reprod Genet. 2017;34(6):795-801.
- Vignini A, Buldreghini E, Nanetti L, Amoroso S, Boscaro M, Ricciardo-Lamonica G, et al. Free thiols in human spermatozoa: are Na+/K+-ATPase, Ca2+-ATPase activities involved in sperm motility through peroxynitrite formation? Reprod Biomed Online. 2009;18(1):132-40.
- Salvador J.M, Inesi G, Rigaud J-L, Mata A.M. Ca2+ transport by reconstituted synaptosomal ATPase is associated with H+ countertransport and net charge displacement. J. Biol Chem. 1998;273(29):18230-4.
- Brini M, Carafoli E. The plasma membrane Ca2+ ATPase and the plasma membrane sodium calcium exchanger cooperate in the regulation of cell calcium. Cold Spring Harb Perspect Biol. 2011;3(2):a004168.
- Schuh K, Cartwright E.J, Jankevics E, Bundschu K, Liebermann J, Williams J.C, et al. Plasma membrane Ca2+ ATPase 4 is required for sperm motility and male fertility. J. Biol Chem. 2004;279(27):28220-6.
- Patel R, Al-Dossary A.A, Stabley D.L, Barone C, Galileo D.S, Strehler E.E, et al. Plasma membrane Ca2+-ATPase 4 in murine epididymis: secretion of splice variants in the luminal fluid and a role in sperm maturation. Biol Reprod. 2013;89(1):6,1-11.
- Williams K, Ford W. Effects of Ca‐ATPase inhibitors on the intracellular calcium activity and motility of human spermatozoa. Int .J. Androl. 2003;26(6):366-75.
- Ranawat P, Kaushik G, Saikia U, Pathak C, Khanduja K. Quercetin impairs the reproductive potential of male mice. Andrologia. 2013;45(1):56-65.
- Sönmez M, Türk G, Çeribaşı S, Çiftçi M, Yüce A, Güvenç M, et al. Quercetin attenuates carbon tetrachloride‐induced testicular damage in rats. Andrologia. 2014;46(8):848-58.
- Liang X, Xia Z, Yan J, Wang Y, Xue S, Zhang X. Quercetin inhibits human sperm functions by reducing sperm [CA 2++] i and tyrosine phosphorylation. Pak .J. Pharm Sci. 2016;29.
- Prasad V, Okunade G.W, Miller M.L, Shull G.E. Phenotypes of SERCA and PMCA knockout mice. Biochem Biophys Res Commun. 2004;322(4):1192-203.
- Poburko D, Santo-Domingo J, Demaurex N. Dynamic regulation of the mitochondrial proton gradient during cytosolic calcium elevations. J. Biol Chem. 2011;286(13):11672-84.
- Boczek T, Lisek M, Ferenc B, Kowalski A, Stepinski D, Wiktorska M, et al. Plasma membrane Ca2+-ATPase isoforms composition regulates cellular pH homeostasis in differentiating PC12 cells in a manner dependent on cytosolic Ca2+ elevations. PLoS One. 2014;9(7):e102352.
- Chalmers S, McCarron J.G. The mitochondrial membrane potential and Ca2+ oscillations in smooth muscle. J. Cell Sci. 2008;121(1):75-85.
- McKenzie E.C, Valberg S.J, Godden S.M, Finno C.J, Murphy M.J. Effect of oral administration of dantrolene sodium on serum creatine kinase activity after exercise in horses with recurrent exertional rhabdomyolysis. Am.J. Vet Res. 2004;65(1):74-9.
- Malli R, Frieden M, Osibow K, Graier W.F. Mitochondria efficiently buffer subplasmalemmal Ca2+ elevation during agonist stimulation. J. Biol Chem. 2003;278(12):10807-15.