The TMEM16A channel mediates the fast polyspermy block in Xenopus laevis

The Journal of General Physiology
Katherine L WozniakAnne E Carlson

Abstract

In externally fertilizing animals, such as sea urchins and frogs, prolonged depolarization of the egg immediately after fertilization inhibits the entry of additional sperm-a phenomenon known as the fast block to polyspermy. In the African clawed frog Xenopus laevis, this depolarization is driven by Ca2+-activated Cl- efflux. Although the prominent Ca2+-activated Cl- currents generated in immature X. laevis oocytes are mediated by X. laevis transmembrane protein 16a (xTMEM16A) channels, little is known about the channels that contribute to the fast block in mature eggs. Moreover, the gamete undergoes a gross transformation as it develops from an immature oocyte into a fertilization-competent egg. Here, we report the results of our approach to identify the Ca2+-activated Cl- channel that triggers the fast block. By querying published proteomic and RNA-sequencing data, we identify two Ca2+-activated Cl- channels expressed in fertilization-competent X. laevis eggs: xTMEM16A and X. laevis bestrophin 2A (xBEST2A). By exogenously expressing xTMEM16A and xBEST2A in axolotl cells lacking endogenous Ca2+-activated currents, we characterize the effect of inhibitors on currents mediated by these channels. None of the inhibitors tested blo...Continue Reading

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Citations

Aug 8, 2018·The Journal of General Physiology·Laurinda A Jaffe
Sep 14, 2019·Proceedings of the National Academy of Sciences of the United States of America·Kuai YuH Criss Hartzell
Aug 21, 2019·Cold Spring Harbor Perspectives in Biology·Takuya WakaiRafael A Fissore
Aug 1, 2020·PLoS Biology·Katherine L WozniakAnne E Carlson
Aug 14, 2019·The Journal of General Physiology·Matthew C Trudeau
Jul 18, 2018·The Journal of General Physiology·Katherine L WozniakAnne E Carlson
Jul 4, 2019·The Journal of Biological Chemistry·Maiwase TemboAnne E Carlson
May 16, 2019·Molecular Reproduction and Development·Katherine L Wozniak, Anne E Carlson
Mar 20, 2019·The Journal of General Physiology·Maiwase Tembo, Anne E Carlson
Aug 29, 2019·Proceedings of the National Academy of Sciences of the United States of America·Anne E Carlson
Dec 30, 2020·Cell·Pablo Ávalos PradoGuillaume Sandoz
Mar 21, 2021·Journal of Molecular Biology·Valeria KalienkovaCristina Paulino
Oct 8, 2021·Molecular Reproduction and Development·Mami WatabeYasuhiro Iwao

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Datasets Mentioned

BETA
SRX1287707

Methods Mentioned

BETA
dissection
PCR
RNA-seq

Software Mentioned

Excel
Photoshop
LAS AF
MONNA
Igor
pClamp
ImageJ
LAS

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