Voltage-gated proton channels exist in the plasma membrane of human oocytes

AbstractSTUDY QUESTIONDo human oocytes express voltage-gated proton channels?SUMMARY ANSWERHuman oocytes exhibit voltage-gated proton currents.WHAT IS KNOWN ALREADYVoltage-gated proton currents have been reported in human sperm, where they contribute to capacitation and motility. No such studies of human oocytes exist.STUDY DESIGN, SIZE, DURATIONVoltage-clamp studies were undertaken using entire oocytes and vesicles derived from oocytes and in excised patches of membrane from oocytes.PARTICIPANTS/MATERIALS, SETTING, METHODSFrozen, thawed human metaphase II oocytes were obtained from material donated to the gamete repository at the Rush Center for Advanced Reproductive Care. Prior to patch clamping, oocytes were warmed and equilibrated. Formation of an electrically tight seal requires exposing bare oolemma. Sections of the zona pellucida (ZP) were removed using a laser, followed by repeated pipetting, to further separate the oocyte from the ZP. Patch-clamp studies were performed using the whole-cell configuration on oocytes or vesicles derived from oocytes, and using inside-out patches of membrane, under conditions optimized to detect voltage-gated proton currents.MAIN RESULTS AND THE ROLE OF CHANCEProton currents are present at significant levels in human oocytes where they exhibit properties similar to those reported in other human cells, as well as those in heterologous expression systems transfected with theHVCN1 gene that codes for the voltage-gated proton channel.LARGE S...
Source: Human Reproduction - Category: Reproduction Medicine Source Type: research