Molecular characterization of voltage-gated potassium channel (Kv) and its importance in functional dynamics in bull spermatozoa

被引:14
作者
Gupta, Rishi Kumar [1 ]
Swain, Dilip Kumar [2 ]
Singh, Vijay [1 ]
Anand, Mukul [2 ]
Choudhury, Soumen [3 ]
Yadav, Sarvajeet [2 ]
Saxena, Atul [1 ]
Garg, Satish Kumar [3 ]
机构
[1] UP Pandit Deendayal Upadhayaya Pashu Chikitsa Vig, Dept Vet Gynaecol & Obstet, Coll Vet Sci & Anim Husb, Mathura 281001, Uttar Pradesh, India
[2] UP Pandit Deendayal Upadhayaya Pashu Chikitsa Vig, Dept Vet Physiol, Coll Vet Sci & Anim Husb, Mathura 281001, Uttar Pradesh, India
[3] UP Pandit Deendayal Upadhayaya Pashu Chikitsa Vig, Dept Vet Pharmacol, Coll Vet Sci & Anim Husb, Mathura 281001, Uttar Pradesh, India
关键词
Voltage gated potassium channel; 4-AP; Sperm motility; Bull; Capacitation; MTP; Volume regulation; APOPTOSIS LIKE CHANGES; ION CHANNELS; VOLUME REGULATION; TYROSINE PHOSPHORYLATION; MALE-FERTILITY; K+ CHANNELS; SPERM; CELLS; SUPPLEMENTATION; QUININE;
D O I
10.1016/j.theriogenology.2018.03.030
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Present study was undertaken to characterize the voltage gated potassium channel (K-v 1.1) in bull spermatozoa using sixty four ejaculates collected from four Hariana bulls. Functional characterization was undertaken using a selective blocker of Kv channel, 4-Aminopyridine (4-AP) while molecular presence of Kv on bull spermatozoa by immunoblotting and indirect immunofluorescence. Three sets of 100 mu L diluted sperm samples namely-negative control (1004 of sperm dilution medium (SDM) containing 10 x 10(6) cells), vehicle control (99 pi of SDM containing 10 x 10(6) cells, and DMSO- 1 mu L.) and 4-AP treatment group (994 of SDM containing 10 x 106 cells, and drug 1 mu L 4-AP) were used in the study. Immunoblotting identified a single band of 56 kDa corresponding to Kvl.1 in Hariana bull spermatozoa. Immunolocalization showed the positive immunoreactivity at head, middle piece and principal piece of the spermatozoa for Kv 1.1. Blocking of Kv using 4-AP resulted in significant (p < 0.05) reduction in sperm progressive motility, per cent capacitated spermatozoa (B-pattern) and acrosome reacted (AR-pattern) spermatozoa, while significant (P < 0.05) increase in per cent swollen spermatozoa. Blocking of Kv channels resulted in significantly (P < 0.05) increased percentage of spermatozoa with lower mitochondria) transmembrane potential. Computer assisted semen analysis (CASA) of motion and kinematic parameters in 4-AP treated spermatozoa indicated reduction in sperm motion parameters like LIN, STR, VSL and VAP and higher ALH, VCL, and BCF indicating hyperactivity of spermatozoa. Based on our findings, it may be concluded that voltage-gated potassium channel (Kv) are present on bull spermatozoa and these are associated with functional dynamics of spermatozoa. However, based on our limited study, it is not possible to deduce that how these channels are associated with induction of hyperactivity. Therefore, further studies are warranted to unravel the mechanistic signaling pathways involved in Kv-mediated alterations in functional dynamics of spermatozoa.(C) 2018 Elsevier Inc. All rights reserved.
引用
收藏
页码:229 / 236
页数:8
相关论文
共 23 条
  • [1] KATP channels in mouse spermatogenic cells and sperm, and their role in capacitation
    Acevedo, JJ
    Mendoza-Lujambio, I
    de la Vega-Beltrán, JL
    Treviño, CL
    Felix, R
    Darszon, A
    [J]. DEVELOPMENTAL BIOLOGY, 2006, 289 (02) : 395 - 405
  • [2] Analysis of the mechanism by which calcium negatively regulates the tyrosine phosphorylation cascade associated with sperm capacitation
    Baker, MA
    Hetherington, L
    Ecroyd, H
    Roman, SD
    Aitken, RJ
    [J]. JOURNAL OF CELL SCIENCE, 2004, 117 (02) : 211 - 222
  • [3] Characterization of potassium channels involved in volume regulation of human spermatozoa
    Barfield, JP
    Yeung, CH
    Cooper, TG
    [J]. MOLECULAR HUMAN REPRODUCTION, 2005, 11 (12) : 891 - 897
  • [4] The effects of putative K+ channel blockers on volume regulation of murine spermatozoa
    Barfield, JP
    Yeung, CH
    Cooper, TG
    [J]. BIOLOGY OF REPRODUCTION, 2005, 72 (05) : 1275 - 1281
  • [5] Breitbart H, 1999, REV REPROD, V2, P165
  • [6] Functional and molecular characterization of voltage gated sodium channel Nav 1.8 in bull spermatozoa
    Chauhan, Dharmendra Singh
    Swain, Dilip Kumar
    Shah, Nadeem
    Yadav, Hanuman Prasad
    Nakade, Udayraj P.
    Singh, Vijay Kumar
    Nigam, Rajesh
    Yadav, Sarvajeet
    Garg, Satish Kumar
    [J]. THERIOGENOLOGY, 2017, 90 : 210 - 218
  • [7] CALCIUM CHANNELS IN THE DEVELOPMENT, MATURATION, AND FUNCTION OF SPERMATOZOA
    Darszon, Alberto
    Nishigaki, Takuya
    Beltran, Carmen
    Trevino, Claudia L.
    [J]. PHYSIOLOGICAL REVIEWS, 2011, 91 (04) : 1305 - 1355
  • [8] Identification of distinct K+ channels in mouse spermatogenic cells and sperm
    Felix, R
    Serrano, CJ
    Treviño, CL
    Muñoz-Garay, C
    Bravo, A
    Navarro, A
    Pacheco, J
    Tsutsumi, V
    Darszon, A
    [J]. ZYGOTE, 2002, 10 (02) : 183 - 188
  • [9] Multi-state, 4-aminopyridine-sensitive ion channels in human spermatozoa
    Gu, Y
    Kirkman-Brown, JC
    Korchev, Y
    Barratt, CLR
    Publicover, SJ
    [J]. DEVELOPMENTAL BIOLOGY, 2004, 274 (02) : 308 - 317
  • [10] The Control of Male Fertility by Spermatozoan Ion Channels
    Lishko, Polina V.
    Kirichok, Yuriy
    Ren, Dejian
    Navarro, Betsy
    Chung, Jean-Ju
    Clapham, David E.
    [J]. ANNUAL REVIEW OF PHYSIOLOGY, VOL 74, 2012, 74 : 453 - 475