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Ion channels from the mouse sperm plasma membrane in planar lipid bilayers

Published online by Cambridge University Press:  26 September 2008

Pedro Labarca
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
Otilia Zapata
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
Carmen Beltrán
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
Alberto Darszon*
Affiliation:
Institute de Biotecnologia, Universidad Nacional Auténoma de México, and Centro de Estudios Cientificos de Santiago and Faculty of Sciences, Universidad de Chile.
*
Dr Alberto Darszon Darszon, Instituto de Biotecnologia, Apartado postal 510–3, Cuernavaca, Morelos, Mexico. Fax: 52-73-172388.

Summary

Fusion of purified mouse sperm plasma membranes to planar lipid bilayers resulted in the insertion of three ion channel types. They could be discerned on the basis of their selectivity, conductance, gating and voltage-dependent properties. The presence of a previously reported large, Ca2+-selective channel was confirmed. Here, it is reported that the Ca21-selective channel from mouse sperm plasmamembrane displayed a pNa+/Pk+ = 1.6 ± 0.2(n=4) and was blocked by micromolar concentrations of ruthenium red. Fusion yielded also a cation-selective channel (PNa+/Pk+ = 2.5±0.3, n=3) with a main open conductance substate of 103 pS and a smaller open substate of 51 PS(600mM K+cis/100 mM Na+trans). The channel inserted into bilayers in two stable fashions: a high-activity mode (open probability = 0.57 ± 0.02, n=3), and a low activity mode (open probability <1%, n=4). In high mode, the channel displayed bursting kinetics and burst length was voltage independent. In addition, a perfectly anion-selective channel, with a slope conductance of 83 PS (600KCI cis/100KCI trans), was identified. It displayed a high, nearly constant open probability (∼0.90)in the 0 to –80 mV range.

Type
Article
Copyright
Copyright © Cambridge University Press 1995

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