Voltage-gated proton channels exist in the plasma membrane of human oocytes.
Voltage-gated proton channels exist in the plasma membrane of human oocytes.
复制标题
电压门控质子通道存在于人类卵母细胞的质膜中。
DOI：
10.1093/humrep/dez178
复制
发表时间：
2019
期刊：
影响因子：
--
通讯作者：
DeCoursey,TE
中科院分区：
文献类型：
--
作者：
Smith,RYa;Morgan,D;Sharma,L;Cherny,VV;Tidswell,N;Molo,MW;DeCoursey,TE
研究问题人类卵母细胞是否表达电压门控质子通道？总结答案人类卵母细胞表现出电压门控质子电流。已知的信息已报道人类精子中存在电压门控质子电流，它们有助于获能和运动。不存在这样的人类卵母细胞研究。研究设计、大小、持续时间使用整个卵母细胞和源自卵母细胞的囊泡以及从卵母细胞切除的膜片进行电压钳研究。参与者/材料、设置、方法冷冻、解冻的人类中期 II 卵母细胞是从捐赠给拉什高级中心配子储存库的材料中获得的 生殖护理。在膜片钳之前，将卵母细胞加热并平衡。形成电密封需要暴露裸露的油膜。使用激光去除透明带 (ZP) 切片，然后重复移液，以进一步将卵母细胞与 ZP 分离。在优化检测电压门控质子电流的条件下，使用卵母细胞或源自卵母细胞的囊泡的全细胞结构，并使用由内而外的膜片进行膜片钳研究。主要结果和机会的作用质子电流在人类卵母细胞中以显着水平存在，它们表现出与其他人类细胞以及转染的异源表达系统中报道的特性相似的特性。 编码电压门控质子通道的 HVCN1 基因。大规模数据/限制，注意原因人类卵母细胞是大细胞，这限制了我们控制细胞内溶液的能力。不能排除通过玻璃化冷冻保存和随后的升温对 HVCN1（HVCN1 基因产物）特性的微妙影响。研究结果的更广泛影响现在可以考虑人类卵母细胞中电压门控质子通道的可能功能。研究资助/竞争利益（S）NIH R35GM126902（TED），Bears Care （DM）。无利益冲突。试用注册号/A。
STUDY 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 theHVCN1gene that codes for the voltage-gated proton channel.LARGE SCALE DATAN/ALIMITATIONS, REASONS FOR CAUTIONHuman oocytes are large cells, which limits our ability to control the intracellular solution. Subtle effects of cryopreservation by vitrification and subsequent warming on properties of HVCN1, theHVCN1gene product, cannot be ruled out.WIDER IMPLICATIONS OF THE FINDINGSPossible functions for voltage-gated proton channels in human oocytes may now be contemplated.STUDY FUNDING/COMPETING INTEREST(S)NIH R35GM126902 (TED), Bears Care (DM). No competing interests.TRIAL REGISTRATION NUMBERN/A.