Pharmacological inhibition of KSper impairs flagellar pH homeostasis of human spermatozoa DOI

Nanxi Ji,

Xiaorong Wang, Xuhui Zeng

et al.

Andrology, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 5, 2024

Abstract Background Sperm‐specific potassium channel (KSper) comprised of pore‐forming subunit SLO3 and auxiliary LRRC52 is importance for sperm fertility. The deficiency KSper in both mice humans resulted severe impairments functions including hyperactivity acrosome reaction. Previous reports suggested that mouse modulated function possibly by affecting intracellular pH (pH i ). However, the precise signaling mechanism human (hKSper) on regulation was largely unclear. Objective To explore regulatory role hKSper flagellar . Materials methods More than 50 donors were recruited during a period 1 year. As reported our previous work, we quantitatively measured employing single‐cell fluorescent recording spermatozoa loaded with indicator pHrodo Red. Three different antagonists clofilium, quinidine, polyclonal antibody (LID1) utilized to evaluate effect inhibition Results Given predominant membrane potential (Em), first detected considerable depolarization (about 25–30 mV) Em evoked clofilium quinidine. Subsequently, it shown values significantly decreased treatment (50 µM, from 7.13 ± 0.11 6.43 0.12), quinidine (500 7.00 6.64 0.08) LID1 (20 µg/mL, 6.98 0.16 6.67 0.22). Moreover, found when pre‐incubated high K + solution (135 mM), acidification abolished. In addition, extracellular substitution N ‐methyl‐D‐glucamine Na abolished induced inhibition. Discussion conclusion Our results demonstrate evokes spermatozoa, suggesting maintenance an important functions.

Language: Английский

Flagellar pH homeostasis mediated by Na+/H+ exchangers regulates human sperm functions through coupling with CatSper and KSper activation DOI
Minzu Liang,

Nanxi Ji,

Jian Song

et al.

Human Reproduction, Journal Year: 2024, Volume and Issue: 39(4), P. 674 - 688

Published: Feb. 14, 2024

Abstract STUDY QUESTION Whether and how do Na+/H+ exchangers (NHEs) regulate the physiological functions of human sperm? SUMMARY ANSWER NHE-mediated flagellar intracellular pH (pHi) homeostasis facilitates activation pH-sensitive, sperm-specific Ca2+ channel (CatSper) K+ (KSper), which subsequently modulate sperm motility, hyperactivation, tyrosine phosphorylation, progesterone (P4)-induced acrosome reaction. WHAT IS KNOWN ALREADY Sperm pHi alkalization is an essential prerequisite for acquisition sperm-fertilizing capacity. Different are strictly controlled by particular regulatory mechanisms. NHEs suggested to H+ efflux. DESIGN, SIZE, DURATION This was a laboratory study that used samples from >50 donors over period 1 year. To evaluate NHE action on function, 5-(N,N-dimethyl)-amiloride (DMA), highly selective inhibitor NHEs, utilized. All experiments were repeated at least five times using different individual or cells. PARTICIPANTS/MATERIALS, SETTING, METHODS By utilizing fluorescent indicator pHrodo Red-AM, we detected alterations in single-cell value sperm. The currents CatSper KSper recorded whole-cell patch-clamp technique. Changes population concentrations ([Ca2+]i) loaded with Fluo 4-AM measured. Membrane potential (Vm) quantitatively examined multimode plate reader after 3,3′-dipropylthiadicarbocyanine iodide 2′,7′-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein, acetoxymethyl ester, respectively. motility parameters assessed computer-assisted semen analysis system. Tyrosine phosphorylation determined immunofluorescence, reaction evaluated Pisum sativum agglutinin-FITC staining. MAIN RESULTS AND THE ROLE OF CHANCE DMA-induced inhibition severely acidified 7.20 ± 0.04 6.38 0.12 (mean SEM), while effect DMA acrosomal less obvious (from 5.90 0.13 5.57 0.12, mean SEM). recordings revealed remarkably suppressed alkalization-induced KSper. As consequence, impairment [Ca2+]i Vm maintenance presence DMA. During capacitation process, pre-treatment 2 h potently decreased pHi, turn kinetic parameters. capacitation-associated functions, including P4-induced reaction, also compromised inhibition. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION vitro study. Caution should be taken when extrapolating these results vivo applications. WIDER IMPLICATIONS FINDINGS important regulators KSper, indispensable fertility, suggesting malfunction could underlying mechanism pathogenesis male infertility. FUNDING/COMPETING INTEREST(S) work supported National Natural Science Foundation China (32271167 81871202 X.Z.), Jiangsu Innovation Entrepreneurship Talent Plan (JSSCRC20211543 Social Development Project Province (No. BE2022765 Society livelihood Nantong City MS22022087 (BK20220608 H.K.). authors have no competing interests declare.

Language: Английский

Citations

6
Direct Effects of Antipsychotics on Potassium Channels DOI

Wenwen Zhuang,

Seo‐Yeong Mun, Won Sun Park

et al.

Biochemical and Biophysical Research Communications, Journal Year: 2025, Volume and Issue: 749, P. 151344 - 151344

Published: Jan. 18, 2025

Language: Английский

Citations

0
Ion channels and transporters involved in calcium flux regulation in mammalian sperm DOI

Claudia Sánchez‐Cárdenas,

Enrique I. Oliver, Julio C. Chávez

et al.

Current topics in developmental biology/Current Topics in Developmental Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Language: Английский

Citations

0
Serotonin–Induced Sperm Hyper‐Motility In Pacific Oyster (Crassostrea Gigas) Associates With K+ Efflux and Membrane Hyperpolarization DOI
Sayyed Mohammad Hadi Alavi, Sepideh Barzegar‐Fallah, Manabu Yoshida

et al.

Journal of Experimental Zoology Part A Ecological and Integrative Physiology, Journal Year: 2025, Volume and Issue: unknown

Published: April 9, 2025

ABSTRACT Serotonin (5‐HT) induces sperm hyper‐motility in bivalves. This process has been suggested to be associated with K + efflux due higher concentrations of ions testicular fluid compared that seawater. hypothesis was supported by inhibition 5‐HT–induced artificial seawater (ASW) containing high extracellular or the presence a voltage‐dependent channel blocker (4‐AP). Here, we studied changes membrane potential elucidate signaling Pacific oyster ( Crassostrea gigas ). Sperm motility partially initiated (48.34 ± 7.80%) ASW, and decreased at 5 min post‐activation p < 0.05). In 10 –5 M 5‐HT, recorded 81.63 3.55%, which remained unchanged within 60 post‐activation. After activation ASW without fluorescence intensity potential‐sensitive fluorescent (DiSC 3 (5)) lower than resting stage, indicating hyperpolarization. Induction hyperpolarization, using valinomycin ‐free (KF‐ASW) could not trigger hyper‐motility, suggesting hyperpolarization itself did induce hyper‐motility. Next, showed efflux. The DiSC (5) increased KF‐ASW 4‐AP, depolarization valinomycin–induced changed subsequent additions KCl, electrochemical gradient resulted retention intracellular ions. Observed 4‐AP Taken together, this study shows

Language: Английский

Citations

0
The essential calcium channel of sperm CatSper is temperature-gated DOI Creative Commons
Dilip Kumar Swain, Citlalli Vergara, Júlia Castro-Arnau

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: April 17, 2025

Abstract The flagellar calcium channel CatSper is essential for male fertility, as it regulates influx to trigger the hyperactive motility required sperm fertilize egg. Precise activation of critical, premature can impair function. While optimal temperature known influence fertilization, its effect on remains unknown. By directly recording from mouse spermatozoa, we reveal that functions a temperature-gated ion channel, with thermal threshold 33.5 °C and coefficient Q 10 5.1. Additionally, show physiological levels spermine reversibly inhibit CatSper’s gating, protecting against activation. Our findings highlight first time presence temperature-gating modality protective role spermine, major component seminal plasma. These results emphasize need maintain testes below 34 fertility advance understanding regulation in fertility.

Language: Английский

Citations

0
Molecular mechanisms of mammalian sperm capacitation, and its regulation by sodium‐dependent secondary active transporters DOI Creative Commons
Gen L. Takei

Reproductive Medicine and Biology, Journal Year: 2024, Volume and Issue: 23(1)

Published: Jan. 1, 2024

Mammalian spermatozoa have to be "capacitated" fertilization-competent. Capacitation is a collective term for the physiological and biochemical changes in that occur within female body. However, regulatory mechanisms underlying capacitation not been fully elucidated.

Language: Английский

Citations

3
The sodium-proton exchangers sNHE and NHE1 control plasma membrane hyperpolarization in mouse sperm DOI Creative Commons

Analia G. Novero,

Paulina Torres‐Rodríguez,

Jose L de La Vega Beltran

et al.

Journal of Biological Chemistry, Journal Year: 2024, Volume and Issue: unknown, P. 107932 - 107932

Published: Oct. 1, 2024

Language: Английский

Citations

2
Hyperglycemia adversely affects critical physiological events related to rat sperm capacitation DOI
Hiram Pacheco-Castillo,

Erika Elena Zagal-Huerta,

Juan José Acevedo‐Fernández

et al.

Biochemical and Biophysical Research Communications, Journal Year: 2024, Volume and Issue: 734, P. 150610 - 150610

Published: Aug. 28, 2024

Language: Английский

Citations

1
Pharmacological inhibition of KSper impairs flagellar pH homeostasis of human spermatozoa DOI

Nanxi Ji,

Xiaorong Wang, Xuhui Zeng

et al.

Andrology, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 5, 2024

Abstract Background Sperm‐specific potassium channel (KSper) comprised of pore‐forming subunit SLO3 and auxiliary LRRC52 is importance for sperm fertility. The deficiency KSper in both mice humans resulted severe impairments functions including hyperactivity acrosome reaction. Previous reports suggested that mouse modulated function possibly by affecting intracellular pH (pH i ). However, the precise signaling mechanism human (hKSper) on regulation was largely unclear. Objective To explore regulatory role hKSper flagellar . Materials methods More than 50 donors were recruited during a period 1 year. As reported our previous work, we quantitatively measured employing single‐cell fluorescent recording spermatozoa loaded with indicator pHrodo Red. Three different antagonists clofilium, quinidine, polyclonal antibody (LID1) utilized to evaluate effect inhibition Results Given predominant membrane potential (Em), first detected considerable depolarization (about 25–30 mV) Em evoked clofilium quinidine. Subsequently, it shown values significantly decreased treatment (50 µM, from 7.13 ± 0.11 6.43 0.12), quinidine (500 7.00 6.64 0.08) LID1 (20 µg/mL, 6.98 0.16 6.67 0.22). Moreover, found when pre‐incubated high K + solution (135 mM), acidification abolished. In addition, extracellular substitution N ‐methyl‐D‐glucamine Na abolished induced inhibition. Discussion conclusion Our results demonstrate evokes spermatozoa, suggesting maintenance an important functions.

Language: Английский

Citations

0