Characterization and expression of SLO1 potassium channels during spermatogenesis inEriocheir sinensis

Abstract

SLO1 potassium channels are pivotal to many aspects of spermatogenic cell. Experiments were conducted to assess physiology and function of SLO1 potassium channels in different developmental stages of spermatogenic cell in Eriocheir sinensis. First, the expression of SLO1 protein was examined via Western blot, RT-PCR, immunohistochemistry and immunofluorescence assays. The results showed that the expression of the SLO1 protein was not uniform in the spermatogenic cells of E. sinensis. Second, whole-cell patch clamp technique was used to record the potassium current of spermatogenic cells and to analyse the electrophysiological characteristics of the potassium channels with the aid of inhibitors. It is proved that the potassium current in E. sinensis germ cells is associated with intracellular Ca2+, and the calcium-activated potassium channel mediated by SLO1 protein is mainly large-conductance Ca2+-activated K+ channels (BKCa). Based on these researches, the cDNA of SLO1 from testis was cloned and sequenced. The SLO1 protein contained domains bound to calcium ions, and the spatial structure formed by its tetramers constituted potassium channels. Phylogenetic analysis revealed that SLO1 was much closer to Scylla paramamosain than other examined species. Finally, iberiotoxin (IbTX) and CdCl2 were used to inhibit the acrosome reaction (AR) induced by A23187 and to explore the role of SLO1 potassium channels in the AR of E. sinensis. The experimental results showed that SLO1 potassium channels were expressed in E. sinensis spermatogenic cells and played an important role in the AR of crab sperm (SP).