HVCN1 Channels Are Relevant for the Maintenance of Sperm Motility: Comparison
Please note this is a comparison between Version 3 by Nora Tang and Version 4 by Nora Tang.

In mammals, sperm capacitation is characterized by a set of physiological changes preparing the male gamete for fertilization, being the intracellular alkalinization a key event. Changes in intracellular pH (pHi) during capacitation are induced by different channels, including HCO3- membrane transporters, Na+-H+ exchangers (NHEs), monocarboxylate transporters (MCTs) and voltage-gated proton channels (HVCN1). HVCN1 channels belong to the superfamily of voltage-gated cation channels; they drive protons more quickly and efficiently than transporters or exchangers do and lead them unidirectionally to the extracellular medium. HVCN1 channels have been identified in human, bull and boar sperm, forming dimers of a molecular weight of 70-73 kDa; however, their action and regulation mechanisms are poorly understood.

A recent study has focused on the physiological role of HVCN1 channels during in vitro capacitation using the pig sperm as a model. This functional approach was carried out pharmacologically through using 2-guanidino benzimidazole (2-GBI), a specific blocker of HVCN1 channels. Sperm samples were incubated in in vitro capacitating medium for 300 min; after 240 min of incubation, progesterone was added to induce sperm hyperactivation and acrosomal exocytosis. To address the physiological role of HVCN1 channels during in vitro capacitation of pig sperm, some samples were incubated in the presence of 2-GBI blocker added at time 0 (Experiment 1). Moreover, and in order to understand the functional relationship between progesterone and HVCN1 channels, 2-GBI blocker was added together with progesterone at 240 min of incubation, in a second group of samples (Experiment 2). Sperm viability, sperm motility and kinematics, acrosomal exocytosis, membrane lipid disorder, intracellular calcium levels of the sperm head and tail, and mitochondrial membrane potential were evaluated after 0, 60, 120, 180, 240, 250, 270 and 300 min of incubation.

The results obtained showed that HVCN1 channels are essential for the maintenance of viability, motility and kinematics of pig sperm during in vitro capacitation and progesterone-induced acrosomal exocytosis. While a close relationship between HVCN1 activation and mitochondrial membrane potential was observed, HVCN1 channels were not found to be involved in the regulation of Ca2+ influx to the sperm tail. Despite further research being necessary, HVCN1 activation could also modulate Ca2+ entrance to the sperm head and prevent premature acrosomal exocytosis during in vitro capacitation of pig sperm.

  • In vitro capacitation
  • mammal sperm
  • HVCN1 channels
  • sperm motility
  • 2-GBI
  • progesterone

1. IDefintroductition

In mammals, sperm capacitatracellion is characterized by a set of physiological changes preparing the male gamete for fertilization [1]. These physiological changes include increased beat frequency and curvilar alinear motility, intracellular pH (pHi) alkalinization durin, rise in intracellular calcium levels, augmented membrane lipid disorder, plasma membrane hyperpolarization, and tyrosine phosphorylation of specific proteins. All these events are required for further acrosomal exocytosis and sperm hyperactivation [1][2][3][4][5][6][7][8]. Recent evidences support the relevance of ion channels in orchestrating the sequencapae of events associated to sperm capacitation [5][7][9]. However, differences induce the content and physiological action of ion channels exist between mammalian species, thus suggesting a species-specific mechanism of sperm capacitation regulation [5][7][9][10][11].

 

Intracellular alkalinization during capacitation is induced by different channels, including HCO3- membrane transporters, Na+-H+ exchangers (NHEs), monocarboxylate transporters (MCTs) and voltage-gated proton channels (HVCN1) [112][213], despite their specific role in pHi regulation being unclear [112][213][314][415]. Taking into account the relevance of sperm pHi alkalinization for oocyte fertilization, the knowledge of the regulation mechanisms of sperm pHi would provide new insights on the etiology and treatment of male infertility.

2. DevelIntroductiopment

HVCN1 channels belong to the superfamily of voltage-gated cation channels [16,17]; they are composed of two subunits, each containing a proton-permeable voltage-sensing domain (VSD), joined by a coiled-coil domain in the C-terminus [415][516][617]. HVCN1 channels differ from other voltage-gated ion channels because they do not have a pore-forming domain; the proton pathway locates within the VSD, so each subunit has its own pore and can function independently [415][718][819][920]. HVCN1 channels drive protons more quickly and efficiently than transporters or exchangers do, and lead them unidirectionally to the extracellular medium [10].

Interestingly, not only does HVCN1 gating rely on membrane voltage, but also on the pH difference (DpH) across the plasma membrane [1121]. Therefore, when changes in intracellular (pHi) and extracellular pH (pHo) are not associated to changes in DpH (pHo – pHi = 0), the voltage dependence of HVCN1-activation is low; conversely, when changes result in DpH>0 or DpH<0 its voltage activation is high [1222]. Therefore, the voltage activation threshold of HVCN1 channels is dependent on the DpH across the plasma membrane [1323].

HVCN1 channels have an essential regulating role in several cell types [1424]. They have been identified in human, bull and boar sperm, forming dimers of a molecular weight of 70-73 kDa; interestingly, these channels are not present in mouse sperm [1121][1323]. HVCN1 channels are implicated in the activation of sperm motility after ejaculation and showed enhanced activity in capacitated spermatozoa [1121][1323], despite their action and regulation mechanisms being poorly understood.

3. Data, Model, Applications and Influences

In a recent study, we investigated the physiological role of HVCN1 channels during in vitro capacitation using the pig sperm as a model [1525]. This functional approach was carried out pharmacologically through using 2-guanidino benzimidazole (2-GBI), a specific blocker of HVCN1channels that inhibits their proton conductance via binding the intracellular VSD domain in the opened channel [10][617]. Sperm samples were incubated in in vitro capacitating medium for 300 min; after 240 min of incubation, progesterone was added to induce sperm hyperactivation and acrosomal exocytosis. To address the physiological role of HVCN1 channels during in vitro capacitation of pig sperm, some samples were incubated in the presence of 2-GBI blocker at 1, 5 or 10 mM added at time 0 (Experiment 1). Moreover, and in order to understand the functional relationship between progesterone and HVCN1 channels, 2-GBI blocker was added together with progesterone at 240 min of incubation, in a second group of samples (Experiment 2) [1525]. Sperm viability, sperm motility and kinematics, acrosomal exocytosis, membrane lipid disorder, intracellular calcium levels of the sperm head and tail, and mitochondrial membrane potential were evaluated after 0, 60, 120, 180, 240, 250, 270 and 300 min of incubation [1525].

Our approach demonstrated that HVCN1 channels are essential for the maintenance of sperm viability and motility during in vitro capacitation. In both experiments, HVCN1 blockage lead to a decreased progressive sperm motility, which manifested associated with an impaired sperm kinematics, mainly due to alterations in sperm velocity and linearity, and a reduction in mitochondrial membrane potential [1525]. In bull [112] and human sperm [169], HVCN1 channels are considered to be essential for the activation of progressive motility after ejaculation and in the regulation of hypermotility during capacitation. In both non-capacitated and capacitated human sperm, the intracellular alkalinization is critical for sperm motility activation, since it triggers the sperm-specific soluble adenylyl cyclase/protein kinase A pathway (sAC/cAMP/PKA), which stimulates cell metabolism and propels the axoneme [169]. In humans, HVCN1 activity is higher in capacitated than in non-capacitated sperm due the phosphorylation of the channel during capacitation [169][1323]; the phosphorylation of this channel during sperm capacitation has not been demonstrated in other mammal species. The reduction in mitochondrial membrane potential after blockage suggest that HVCN1 channels are essential for pig sperm to increase their oxidative phosphorylation during sperm capacitation and acrosomal exocytosis induced by progesterone [1525]. In agreement with our results, Musset et al. [1726] reported that HVCN1 channels are involved in the generation of superoxide radicals in human sperm, which are reactive oxygen species resulting from the activity of the mitochondrial membrane chain. In humans, alterations in mitochondrial membrane potential observed after HVCN1 inhibition are also associated to the decrease of sperm viability [1827].

During capacitation, HVCN1 channels have been suggested to induce hypermotility through the cAMP/PKA pathway in bull sperm [112], and via activating CatSper channels in human sperm [169][1323][1827][1928]. In capacitated pig sperm, a clear functional relationship between HVCN1 activation and Ca2+ entrance to the sperm tail was not observed, thus suggesting that while HVCN1 channels are essential for pig sperm motility during in vitro capacitation, they do not appear to be crucial in the regulation of Ca2+ influx [1525]. Despite further research being required, we hypothesize that Ca2+ entrance to the flagellum in capacitated pig sperm does not rely upon the activity of HVCN1 channels, but rather depend a direct and non-direct effect of progesterone on Ca2+ channels [1525].

In rodents [2029], cattle [2130] and humans [169], intracellular alkalinization has been correlated with the activation CatSper channels and the rise in intracellular Ca2+ levels during sperm capacitation. Moreover, in human sperm, HVCN1 and CatSper channels are localized in the same plasma membrane domains thus evidencing their strong functional relationship. In our study, however, the absence of a strong relationship between HVCN1 activity and Ca2+ influx to the sperm tail suggests that not only is intracellular alkalinization regulated by HVCN1 channels in pig sperm, but also by other H+ transporters [1525]. Nevertheless, further research in necessary in order to identify and localize the channels implicated in pHi regulation and their functional relationship with calcium channels.

Little data exist about the effects of HVCN1 blockage on acrosomal exocytosis in mammalian species. In pig sperm, HVCN1 inhibition had a different effect on acrosomal exocytosis depending on whether the 2-GBI blocker was added at the beginning of the experiment (i.e. 0 min) or after 240 min of incubation [1525]. In sperm incubated with 2-GBI from time 0 (Experiment 1), a premature acrosomal exocytosis was observed, which was associated to high plasma membrane lipid disorder and to increased Ca2+ levels in the sperm head. In these samples, the addition of progesterone acted as a potent acrosomal exocytosis inducer, neither a new rise in the Ca2+ levels of the sperm head nor a further increase in membrane lipid disorder were observed [1525]. In agreement with our results, a close relationship between pHi and cholesterol content of the plasma membrane has been reported in human sperm [2231]. Moreover, recent studies in rodents have demonstrated that the sterol efflux induces a local depolarizing effect on the plasma membrane that may activate different types of transient voltage-gated cation channels, leading to Ca2+ rises and triggering acrosomal exocytosis [2332][2433]. In contrast, when added at 240 min together with progesterone (Experiment 2), 2-GBI blocker had little effect on acrosomal exocytosis and plasma membrane lipid disorder, but it led to reduced Ca2+ levels of the sperm head [1525]. Besides, zinc blocking of HVCN1 channels in capacitated human sperm reduces progesterone-induced acrosomal exocytosis due to the inhibition of CatSper channels, which again underpins the close functional relationship between these two kinds of channels in this species [1827]. In capacitated pig spermatozoa, SLO1 blockage also inhibits progesterone-induced acrosomal exocytosis by reducing Ca2+ entrance to the sperm head [257], whereas the presence of zinc prevents acrosomal membrane modifications associated to acrosomal exocytosis [2634].

The alterations observed after total blockage in Experiment 1 suggest that HVCN1 channels are essential to maintain pig sperm homeostasis during in vitro capacitation and for preventing premature sperm activation [1525]. From Experiment 2, one can conclude that while HVCN1 channels are relevant for the rise in Ca2+ levels after progesterone addition, they are not specifically involved in the mechanism that triggers acrosomal exocytosis [1525]. On the other hand, another interesting finding of our study was that HVCN1 blockage affected differently the intracellular Ca2+ flux to the sperm head and the sperm tail, thereby indicating that the involvement of HVCN1 channels in regulating these two stores is distinct [1525]. These results are in line with previous studies performed in human [2735], mouse [2836] and pig sperm [257].

References

  1. Abhishek Kumar Mishra; Akshay Kumar; Sarvajeet Yadav; Mukul Anand; Brijesh Yadav; Rajesh Nigam; Satish Kumar Garg; Dilip Kumar Swain; Functional insights into voltage gated proton channel (Hv1) in bull spermatozoa.. ThKarl Kerns; Michal Zigo; Erma Z. Drobnis; Miriam Sutovsky; Peter Sutovsky; Zinc ion flux during mammalian sperm capacitation.. Naturer Communicatiogenology ns 2019, 136, 118-130, 10.1016/j.theriogenology.2019.06.015.8, 9, 2061, 10.1038/s41467-018-04523-y.
  2. I. Scott Ramsey; Magdalene M. Moran; Jayhong A. Chong; David E Clapham; A voltage-gated proton-selective channel lacking the pore domain. NaMarta Puigmulé; Anna Fàbrega; Marc Yeste; Sergi Bonet; Elisabeth Pinart; Study of the proacrosin - acrosin system in epididymal, ejaculated and in vitro capacitated boar spermatozoa. Reproductuion, Fertility and De velopment 2006, 440, 1213-1216, 10.1038/nature04700.11, 23, 837-845, 10.1071/rd10345.
  3. M. Sasaki; Masahiro Takagi; Yasushi Okamura; A Voltage Sensor-Domain Protein Is a Voltage-Gated Proton Channel. ScAnna Fàbrega; Marta Puigmulé; Sergi Bonet; Elisabeth Pinart; Epididymal maturation and ejaculation are key events for further in vitro capacitation of boar spermatozoa. Theriogence ology 2006, 312, 589-592, 10.1126/science.1122352.12, 78, 867-877, 10.1016/j.theriogenology.2012.03.039.
  4. Seok-Yong Lee; James Letts; Roderick MacKinnon; Dimeric subunit stoichiometry of the human voltage-dependent proton channel Hv1. PJose Luis De La Vega-Beltran; Claudia Sánchez-Cárdenas; Dario Krapf; Enrique O. Hernández-González; Eva Wertheimer; Claudia L. Treviño; Pablo E Visconti; Alberto Darszon; Mouse Sperm Membrane Potential Hyperpolarization Is Necessary and Sufficient to Prepare Sperm for the Acrosome Reaction*. Jouroceedings of the National Academy of Scienceal of Biological Chemis try 2008, 105, 7692-7695, 10.1073/pnas.0803277105.12, 287, 44384-44393, 10.1074/jbc.M112.393488.
  5. Liang Hong; Medha M. Pathak; Iris H. Kim; Dennis Ta; Francesco Tombola; Voltage-Sensing Domain of Voltage-Gated Proton Channel Hv1 Shares Mechanism of Block with Pore Domains. NChristoph Brenker; Yu Zhou; Astrid Müller; Fabio Andres Echeverry; Christian Trötschel; Ansgar Poetsch; Xiao-Ming Xia; Wolfgang Bönigk; Christopher J. Lingle; U Benjamin Kaupp; Timo Strünker; The Ca2+-activated K+ current of human sperm is mediated by Slo3. eLifeuron 2013, 79, 202, 10.1016/j.neuron.2013.06.031.4, 3, e01438, 10.7554/eLife.01438.
  6. Liang Hong; Iris H. Kim; Francesco Tombola; Molecular determinants of Hv1 proton channel inhibition by guanidine derivatives. PMarc Yeste; J. M. Fernández-Novell; Laura Ramió-Lluch; E. Estrada; L. G. Rocha; J. A. Cebrián‐Pérez; T. Muiño‐Blanco; I. I. Concha; A. Ramirez; Joan Enric Rodríguez‐Gil; Intracellular calcium movements of boar spermatozoa during ‘in vitro’ capacitation and subsequent acrosome exocytosis follow a multiple-storage place, extracellular calcium-dependent model. Androceedinlogs of the National Academy of Sciences 2014, 111, 9971-9976, 10.1073/pnas.1324012111.5, 3, 729-747, 10.1111/andr.12054.
  7. S. B. Long; Crystal Structure of a Mammalian Voltage-Dependent Shaker Family K+ Channel. Science 2005, 309, 897-903, 10.1126/science.1116269.Yeste, M.; Llavanera, M.; Pérez, G.; Scornik, F.; Puig-Parri, J.; Brugada, R.; Bonet, S.; Pinart, E. Elucidating the role of K+ channels during in vitro capacitation of boar spermatozoa: Do SLO1 channels play a crucial role? Int. J. Mol. Sci. 2019, 20.
  8. Francesco Tombola; Maximilian H. Ulbrich; Ehud Y. Isacoff; The Voltage-Gated Proton Channel Hv1 Has Two Pores, Each Controlled by One Voltage Sensor. NeArturo Matamoros-Volante; Claudia L. Treviño; Capacitation-associated alkalization in human sperm is differentially controlled at the subcellular level. nuron ll 2008, 58, 546-56, 10.1016/j.neuron.2008.03.026.19, null, 763987, 10.1101/763987.
  9. Santiago Rebolledo; Feng Qiu; H Peter Larsson; Molecular structure and function of Hv1 channels. WilPolina V Lishko; Inna L. Botchkina; Andriy Fedorenko; Yuriy Kirichok; Acid Extrusion from Human Spermatozoa Is Mediated by Flagellar Voltage-Gated Proton Channel. Cey Interdisciplinary Reviews: Membrane Transport and Signaling 2012, 0, 1, 763-777, 10.1002/wmts.49.40, 327-337, 10.1016/j.cell.2009.12.053.
  10. Andreas Rennhack; Elena Grahn; U. Benjamin Kaupp; Thomas K. Berger; Photocontrol of the Hv1 Proton Channel. ACS Chemical Biology 2017, 12, 2952-2957, 10.1021/acschembio.7b00523.
  11. Thomas K. Berger; David M. Fußhöller; Normann Goodwin; Wolfgang Bönigk; Astrid Müller; Nasim Dokani Khesroshahi; Christoph Brenker; Dagmar Wachten; Eberhard Krause; U. Benjamin Kaupp; Timo Strünker; Post‐translational cleavage of Hv1 in human sperm tunes pH‐ and voltage‐dependent gating. ThRaquel Bernardino; David Carrageta; Mário Sousa; Marco G. Alves; Pedro F. Oliveira; pH and male fertility: making sense on pH homeodynamics throughout the male reproductive tract. Cellular Journal of Phyand Molecular Life Sciencesiology 2017, 595, 1533-1546, 10.1113/JP273189.9, 76, 3783-3800, 10.1007/s00018-019-03170-w.
  12. V V Cherny; V S Markin; T E DeCoursey; The voltage-activated hydrogen ion conductance in rat alveolar epithelial cells is determined by the pH gradient. JouAbhishek Kumar Mishra; Akshay Kumar; Sarvajeet Yadav; Mukul Anand; Brijesh Yadav; Rajesh Nigam; Satish Kumar Garg; Dilip Kumar Swain; Functional insights into voltage gated proton channel (Hv1) in bull spermatozoa.. Thernal iof General Physigenology 201995, , 105, 861-896.36, 118-130, 10.1016/j.theriogenology.2019.06.015.
  13. Polina V. Lishko; Yuriy Kirichok; The role of Hv1 and CatSper channels in sperm activation. The JoI. Scott Ramsey; Magdalene M. Moran; Jayhong A. Chong; David E Clapham; A voltage-gated proton-selective channel lacking the pore domain. Naturnal of Physiology e 2010, 588, 4667-4672, 10.1113/jphysiol.2010.194142.6, 440, 1213-1216, 10.1038/nature04700.
  14. Ruiming Zhao; Kelleigh Kennedy; Gerardo A. De Blas; Gerardo Orta; Martín A. Pavarotti; Rodolfo J. Arias; José Luis De La Vega-Beltrán; Qufei Li; Hui Dai; Eduardo Perozo; Luis S. Mayorga; Alberto Darszon; Steve A.N. Goldstein; Role of human Hv1 channels in sperm capacitation and white blood cell respiratory burst established by a designed peptide inhibitor. Proceedings of the National Academy of M. Sasaki; Masahiro Takagi; Yasushi Okamura; A Voltage Sensor-Domain Protein Is a Voltage-Gated Proton Channel. Sciences 2018, 06, 3115, E11847-E11856, 10.1073/pnas.1816189115.2, 589-592, 10.1126/science.1122352.
  15. E Pinart; R Camps; M D Briz; S Bonet; Sperm quality in spontaneous unilateral abdominal cryptorchid boars.. ThSeok-Yong Lee; James Letts; Roderick MacKinnon; Dimeric subunit stoichiometry of the human voltage-dependent proton channel Hv1. Proceedings of Internthe National Journal of Developmental Biology 1996, 2Academy of Sciences 2008, 1, E3255.05, 7692-7695, 10.1073/pnas.0803277105.
  16. Polina V Lishko; Inna L. Botchkina; Andriy Fedorenko; Yuriy Kirichok; Acid Extrusion from Human Spermatozoa Is Mediated by Flagellar Voltage-Gated Proton Channel. CLiang Hong; Medha M. Pathak; Iris H. Kim; Dennis Ta; Francesco Tombola; Voltage-Sensing Domain of Voltage-Gated Proton Channel Hv1 Shares Mechanism of Block with Pore Domains. Nell uron 2010, 140, 327-337, 10.1016/j.cell.2009.12.053.3, 79, 202, 10.1016/j.neuron.2013.06.031.
  17. Musset, B.; Clark, R.A.; DeCoursey, T.E.; Petheo, G.L.; Geiszt, M.; Chen, Y.; Cornell, J.E.; Eddy, C.A.; Brzyski, R.G.; El Jamali, A. NOX5 in human spermatozoa: Expression, function, and regulation. J. Biol. Chem. 2012, 287, 9376–9388.Liang Hong; Iris H. Kim; Francesco Tombola; Molecular determinants of Hv1 proton channel inhibition by guanidine derivatives. Proceedings of the National Academy of Sciences 2014, 111, 9971-9976, 10.1073/pnas.1324012111.
  18. Sara Keshtgar; Hamideh Ghanbari; Esmaeel Ghani; Seyed Mostafa Shid Moosavi; Effect of CatSper and Hv1 Channel Inhibition on Progesterone Stimulated Human Sperm. Journal of reproduS. B. Long; Crystal Structure of a Mammalian Voltage-Dependent Shaker Family K+ Channel. Sction & infencertility 197 200, 15, 309, 133-139., 897-903, 10.1126/science.1116269.
  19. Hamideh Ghanbari; Sara Keshtgar; Hamid Reza Zare; Behrouz Gharesi-Fard; Inhibition of CatSper and Hv1 Channels and NOX5 Enzyme Affect Progesterone-Induced Increase of Intracellular Calcium Concentration and ROS Generation in Human Sperm.. Iranian joFrancesco Tombola; Maximilian H. Ulbrich; Ehud Y. Isacoff; The Voltage-Gated Proton Channel Hv1 Has Two Pores, Each Controlled by One Voltage Sensor. Neurnal of medical sciences 20019, 44, 127-134.8, 58, 546-56, 10.1016/j.neuron.2008.03.026.
  20. Yuriy Kirichok; Betsy Navarro; David E Clapham; Whole-cell patch-clamp measurements of spermatozoa reveal an alkaline-activated Ca2+ channel. NSantiago Rebolledo; Feng Qiu; H Peter Larsson; Molecular structure and function of Hv1 channels. Wiley Interdisciplinatury Reviews: Membrane Transport and Signaling 2006, 439, 737-740, 10.1038/nature04417.12, 1, 763-777, 10.1002/wmts.49.
  21. Becky Marquez; Susan S. Suarez; Bovine Sperm Hyperactivation Is Promoted by Alkaline-Stimulated Ca2+ Influx1. BiThomas K. Berger; David M. Fußhöller; Normann Goodwin; Wolfgang Bönigk; Astrid Müller; Nasim Dokani Khesroshahi; Christoph Brenker; Dagmar Wachten; Eberhard Krause; U. Benjamin Kaupp; Timo Strünker; Post‐translational cleavage of Hv1 in human sperm tunes pH‐ and voltage‐dependent gating. The Journalogy of Reproduction of Physiology 20017, 76, 660-665, 10.1095/biolreprod.106.055038., 595, 1533-1546, 10.1113/JP273189.
  22. N L Cross; P Razy-Faulkner; Control of human sperm intracellular pH by cholesterol and its relationship to the response of the acrosome to progesterone.. BiV V Cherny; V S Markin; T E DeCoursey; The voltage-activated hydrogen ion conductance in rat alveolar epithelial cells is determined by the pH gradient. Journalogy of Reproduction of General Physiology 1997, 5, 1056, 1169–1174., 861-896.
  23. Roy Cohen; Danielle E. Buttke; Atsushi Asano; Chinatsu Mukai; Jacquelyn L. Nelson; Ngjun Ren; Richard J. Miller; Moshe Cohen-Kutner; Daphne Atlas; Alexander J. Travis; Lipid modulation of calcium flux through CaV2.3 regulates acrosome exocytosis and fertilization.. DPolina V. Lishko; Yuriy Kirichok; The role of Hv1 and CatSper channels in sperm activation. Thevel Jopmental Cell urnal of Physiology 2014, 20, 588, 310-21, 10.1016/j.devcel.2014.01.005., 4667-4672, 10.1113/jphysiol.2010.194142.
  24. Roy Cohen; Chinatsu Mukai; Alexander J. Travis; Lipid Regulation of Acrosome Exocytosis. FaRuiming Zhao; Kelleigh Kennedy; Gerardo A. De Blas; Gerardo Orta; Martín A. Pavarotti; Rodolfo J. Arias; José Luis De La Vega-Beltrán; Qufei Li; Hui Dai; Eduardo Perozo; Luis S. Mayorga; Alberto Darszon; Steve A.N. Goldstein; Role of human Hv1 channels in sperm capacitation and white blood cell respiratory burst established by a designed peptide inhibitor. Proctors Influeencing Mammalian Kidney Development: Implications for Health in Adult Lifdings of the National Academy of Science s 2016, 220, 107-127, 10.1007/978-3-319-30567-7_6.8, 115, E11847-E11856, 10.1073/pnas.1816189115.
  25. Yeste, M.; Llavanera, M.; Pérez, G.; Scornik, F.; Puig-Parri, J.; Brugada, R.; Bonet, S.; Pinart, E. Elucidating the role of K+ channels during in vitro capacitation of boar spermatozoa: Do SLO1 channels play a crucial role? Int. J. Mol. Sci. 2019, 20.E Pinart; R Camps; M D Briz; S Bonet; Sperm quality in spontaneous unilateral abdominal cryptorchid boars.. The International Journal of Developmental Biology 1996, 21, E3255.
  26. Karl Kerns; Momal Sharif; Michal Zigo; Wei Xu; Lauren Hamilton; Miriam Sutovsky; Mark Ellersieck; Erma Drobnis; Nicolai V Bovin; Richard Oko; David Miller; Peter Sutovsky; Sperm Cohort-Specific Zinc Signature Acquisition and Capacitation-Induced Zinc Flux Regulate Sperm-Oviduct and Sperm-Zona Pellucida Interactions. International Journal of Molecular Sciences 2020, 21, 2121, 10.3390/ijms21062121.Musset, B.; Clark, R.A.; DeCoursey, T.E.; Petheo, G.L.; Geiszt, M.; Chen, Y.; Cornell, J.E.; Eddy, C.A.; Brzyski, R.G.; El Jamali, A. NOX5 in human spermatozoa: Expression, function, and regulation. J. Biol. Chem. 2012, 287, 9376–9388.
  27. Linda Lefièvre; Katherine Nash; Steven Mansell; Sarah Costello; Emma Punt; Joao Correia; Jennifer Morris; J. C. Kirkman-Brown; Stuart Wilson; Christopher Lr Barratt; Steve Publicover; 2-APB-potentiated channels amplify CatSper-induced Ca2+ signals in human sperm. BiSara Keshtgar; Hamideh Ghanbari; Esmaeel Ghani; Seyed Mostafa Shid Moosavi; Effect of CatSper and Hv1 Channel Inhibition on Progesterone Stimulated Human Sperm. Jochemicurnal Journaof reproduction & infertil 2ity 197012, 448, 189-200, 10.1042/bj20120339., 19, 133-139.
  28. Long-Fei Li; Cheng Xiang; Ya-Bing Zhu; Kai-Rong Qin; Modeling of progesterone-induced intracellular calcium signaling in human spermatozoa. JHamideh Ghanbari; Sara Keshtgar; Hamid Reza Zare; Behrouz Gharesi-Fard; Inhibition of CatSper and Hv1 Channels and NOX5 Enzyme Affect Progesterone-Induced Increase of Intracellular Calcium Concentration and ROS Generation in Human Sperm.. Iranian journal of Thmeoretical Biology dical sciences 2014, 351, 58-66, 10.1016/j.jtbi.2014.02.026.9, 44, 127-134.
  29. Yuriy Kirichok; Betsy Navarro; David E Clapham; Whole-cell patch-clamp measurements of spermatozoa reveal an alkaline-activated Ca2+ channel. Nature 2006, 439, 737-740, 10.1038/nature04417.
  30. Becky Marquez; Susan S. Suarez; Bovine Sperm Hyperactivation Is Promoted by Alkaline-Stimulated Ca2+ Influx1. Biology of Reproduction 2007, 76, 660-665, 10.1095/biolreprod.106.055038.
  31. N L Cross; P Razy-Faulkner; Control of human sperm intracellular pH by cholesterol and its relationship to the response of the acrosome to progesterone.. Biology of Reproduction 1997, 56, 1169–1174.
  32. Roy Cohen; Danielle E. Buttke; Atsushi Asano; Chinatsu Mukai; Jacquelyn L. Nelson; Ngjun Ren; Richard J. Miller; Moshe Cohen-Kutner; Daphne Atlas; Alexander J. Travis; Lipid modulation of calcium flux through CaV2.3 regulates acrosome exocytosis and fertilization.. Developmental Cell 2014, 28, 310-21, 10.1016/j.devcel.2014.01.005.
  33. Roy Cohen; Chinatsu Mukai; Alexander J. Travis; Lipid Regulation of Acrosome Exocytosis. Factors Influencing Mammalian Kidney Development: Implications for Health in Adult Life 2016, 220, 107-127, 10.1007/978-3-319-30567-7_6.
  34. Karl Kerns; Momal Sharif; Michal Zigo; Wei Xu; Lauren Hamilton; Miriam Sutovsky; Mark Ellersieck; Erma Drobnis; Nicolai V Bovin; Richard Oko; David Miller; Peter Sutovsky; Sperm Cohort-Specific Zinc Signature Acquisition and Capacitation-Induced Zinc Flux Regulate Sperm-Oviduct and Sperm-Zona Pellucida Interactions. International Journal of Molecular Sciences 2020, 21, 2121, 10.3390/ijms21062121.
  35. Linda Lefièvre; Katherine Nash; Steven Mansell; Sarah Costello; Emma Punt; Joao Correia; Jennifer Morris; J. C. Kirkman-Brown; Stuart Wilson; Christopher Lr Barratt; Steve Publicover; 2-APB-potentiated channels amplify CatSper-induced Ca2+ signals in human sperm. Biochemical Journal 2012, 448, 189-200, 10.1042/bj20120339.
  36. Long-Fei Li; Cheng Xiang; Ya-Bing Zhu; Kai-Rong Qin; Modeling of progesterone-induced intracellular calcium signaling in human spermatozoa. Journal of Theoretical Biology 2014, 351, 58-66, 10.1016/j.jtbi.2014.02.026.
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