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).

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Manuscript accepted on :February 10, 2018
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Silvia W Lestari^1,4, Manggiasih D. Larasati², Indra G. Mansur^1,4 and Ria Margiana^3,4

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: finallysilvia@gmail.com

DOI : https://dx.doi.org/10.13005/bpj/1388

Abstract

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.

Keywords

Ca²⁺ATPase Male Infertility; Na⁺ K⁺-ATPase; Sperm Motility;

Introduction

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

Table 2: Mechanism of Na⁺ K⁺-ATPase in sperm motility

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

Table 4: Mechanism of Ca²⁺-ATPase in sperm motility

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.

Conclusion

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.

Acknowledgment

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 Source

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.

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