Cellular Mechanism Underlying Hydrogen Sulfide Mediated Epithelial K⁺ Secretion in Rat Epididymis

As a novel gasotransmitter, hydrogen sulfide (H₂S) elicits various physiological actions including smooth muscle relaxation and promotion of transepithelial ion transport. However, the pro-secretory function of H₂S in the male reproductive system remains largely unclear. The aim of this study is to elucidate the possible roles of H₂S in modulating rat epididymal intraluminal ionic microenvironment essential for sperm storage. The results revealed that endogenous H₂S-generating enzymes cystathionine β-synthetase (CBS) and cystathionine γ-lyase (CSE) were both expressed in rat epididymis. CBS located predominantly in epithelial cells whilst CSE expressed primarily in smooth muscle cells. The relative expression level of CBS and CSE escalated from caput to cauda regions of epididymis, which was paralleled to the progressively increasing production of endogenous H₂S. The effect of H₂S on epididymal epithelial ion transportation was investigated using short-circuit current (I _SC), measurement of intracellular ion concentration and in vivo rat epididymal microperfusion. Our data showed that H₂S induced transepithelial K⁺ secretion via adenosine triphosphate-sensitive K⁺ (K_ATP) channel and large conductance Ca²⁺-activated K⁺ (BK_Ca) channel. Transient receptor potential vanilloid 4 (TRPV4) channel-mediated Ca²⁺ influx was implicated in the activation of BK_Ca channel. In vivo studies further demonstrated that H₂S promoted K⁺ secretion in rat epididymal epithelium. Inhibition of endogenous H₂S synthesis caused a significant decrease in K⁺ concentration of cauda epididymal intraluminal fluid. Moreover, our data demonstrated that high extracellular K⁺ concentration actively depressed the motility of cauda epididymal sperm in a pH-independent manner. Collectively, the present study demonstrated that H₂S was vital to the formation of high K⁺ concentration in epididymal intraluminal fluid by promoting the transepithelial K⁺ secretion, which might contribute to the maintenance of the cauda epididymal sperm in quiescent dormant state before ejaculation.