Volume regulation of mature and immature spermatozoa in a primate model, and possible ion channels involved

Abstract

Background: Human ejaculated sperm undergo volume regulation, and swollen cells fail to penetrate mucus. Study of an infertile mouse model indicates maturation of volume regulation mechanism in the epididymis. Methods: Sperm from the ejaculate and three regions of the epididymis of the cynomolgus monkey (Macaca fascicularis) were dispersed in BWW medium and changes in the cell volume and kinematics, and their responses to ion channel blockers, were monitored by flow cytometry and motion analysis. Results: Initially swollen cauda epididymidal spermatozoa regained their original volume within 20 min, but not in the presence of 0.25 mM quinine. Corpus epididymidal spermatozoa underwent such regulatory volume decrease (RVD) to a lesser extent, with a similar response to quinine. Caput sperm showed no swelling throughout incubation. The chloride channel inhibitor NPPB also caused swelling of cauda spermatozoa and both quinine and NPPB decreased the efficiency of forward progression. RVD of ejaculated spermatozoa was inhibited by the K+ channel blockers quinine and 4-aminopyridine (4-AP) but not by tetraethylammonium, Ba2+ or Gd3+, or the specific potassium channel blockers charybdotoxin, margatoxin, dendrotoxin, apamin, glybenclamide or clofilium. Quinine and 4-AP also altered ejaculated sperm kinematics as reported in human ejaculated spermatozoa. Conclusions: Quinine- and 4-AP-sensitive (implying K+) and NPPB-sensitive (implying Cl-) channels are involved in RVD of primate sperm, which develop this volume regulatory ability in the epididymis. © European Society of Human Reproduction and Embryology 2004; all rights reserved.