Iranian Journal of Medical Sciences
Iranian Journal of Medical Sciences

Inhibition of CatSper and Hv1 Channels and NOX5 Enzyme Affect Progesterone-Induced Increase of Intracellular Calcium Concentration and ROS Generation in Human Sperm

Document Type : Original Article(s)

Authors

Hamideh Ghanbari 1
Sara Keshtgar 1
Hamid Reza Zare 2
Behrouz Gharesi-Fard 3

1 Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

2 Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

3 Department of Immunology, Infertility Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

https://doi.org/10.30476/ijms.2019.44513
Abstract
Background: Normal sperm function depends on appropriate intracellular calcium (Cai2+) and reactive oxygen species (ROS) levels. Calcium activates NADPH oxidase-5 (NOX5) that leads to ROS generation. The calcium channel of sperm (CatSper) is activated by progesterone and intracellular alkalization. Herein, the interactive role of CatSper, Hv1 channels, and NOX5 enzyme on Cai2+ and ROS generation in human sperm is investigated. Methods: The present laboratory in vitro study was carried out in the School of Medicine, Shiraz University of Medical Sciences (Shiraz, Iran) during 2016. Normal semen samples (n=15) were washed and diluted to 20×106 sperm/mL. The diluted samples were divided into 16 groups containing Ham’s F-10 (the control group), 2 µM NNC (CatSper inhibitor), 1 mM ZnCl2 (Hv1 inhibitor), 1 µM DPI (NOX5 inhibitor), NNC+Zn, NNC+DPI, and NNC+Zn+DPI. The other 8 groups were the same as the above except that they contained 1 µM progesterone. Cell viability and Cai2+ were analyzed by flou-3 AM probe and PI staining, respectively, using flow cytometric method. ROS generation was assessed by chemiluminescence method. Statistical analysis was performed using the one-way ANOVA followed by Tukey’s test. P values

Keywords

CATSPERB protein, human
Hv1 channel, human
NADPH oxidase 5
Progesterone
Spermatozoa
Iranian Journal of Medical Sciences
Volume 44, Issue 2 - Serial Number 2
98
March 2019
Pages 127-134


  • Received 09 May 2017
  • Revised 22 July 2019
  • Accepted 10 March 2019

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PDF Download 3,505

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