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A hyperpolarization-activated ion current of amphibian oocytes

  • Ion Channels, Receptors and Transporters
  • Published:
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Abstract

A comparative analysis of a hyperpolarization-activated ion current present in amphibian oocytes was performed using the two-electrode voltage-clamp technique in Xenopus laevis, Xenopus tropicalis, and Ambystoma mexicanum. This current appears to be driven mainly by Cl− ions, is independent of Ca2+, and is made evident by applying extremely negative voltage pulses; it shows a slow activating phase and little or no desensitization. The pharmacological profile of the current is complex. The different channel blocker used for Cl−, K+, Na+ and Ca2+ conductances, exhibited various degrees of inhibition depending of the species. The profiles illustrate the intricacy of the components that give rise to this current. During X. laevis oogenesis, the hyperpolarization-activated current is present at all stages of oocytes tested (II–VI), and the amplitude of the current increases from about 50 nA in stage I to more than 1 μA in stage VI; nevertheless, there was no apparent modification of the kinetics. Our results suggest that the hyperpolarization-activated current is present both in order Anura and Urodela oocytes. However, the electrophysiological and pharmacological characteristics are quite perplexing and seem to suggest a mixture of ionic conductances that includes the activation of both anionic and cationic channels, most probably transiently opened due to the extreme hyperpolarizion of the plasma membrane. As a possible mechanism for the generation of the current, a kinetic model which fits the data suggests the opening of pores in the plasma membrane whose ion selectivity is dependent on the extracellular Cl− concentration. The extreme voltage conditions could induce the opening of otherwise latent pores in plasma membrane proteins (i.e., carriers), resembling the ´slippage´ events already described for some carriers. These observations should be valuable for other groups trying to express cloned, voltage-dependent ion channels in oocytes of amphibian in which hyperpolarizing voltage pulses are applied to activate the channels.

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Abbreviations

4-AP:

4-Aminopyridine

9-AC:

Anthracene-9-carboxylic acid

BAPTA:

1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid

DIDS:

4,4′-Diisothiocyanatostilbene, 2,2′-disulfonic acid disodium salt hydrate

DMSO:

Dimethyl sulfoxide

HEPES:

N-2-Hydroxyethylpiperazine-N′-2-ethanosulfonic acid

MES:

2-(N-Morpholino)ethanesulfonic acid

MS-222:

3-Aminobenzoic acid methyl ester

NFA:

Niflumic acid

NPPB:

5-Nitro-2(3-phenylpropylamino)benzoic acid

TEA:

Tetraethylammonium

TTX:

Tetrodotoxin

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Acknowledgments

We thank E. Ruiz Alcibar and E. Espino Saldaña for technical support. This work was supported by grants from CONACYT (101851), PAPIIT-UNAM (IN202609 and IN205308). DBSS received a fellowship from CONACYT and was supported by Posgrado en Ciencias Biomédicas-UNAM. Dr. D. D. Pless edited the original manuscript and we are grateful for her critical reviews.

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  1. L. D. Ochoa-de la Paz

    Present address: Departamento de Investigación Biomédica, Facultad de Medicina, Universidad Autónoma de Querétaro, Querétaro, Qro, CP 76176, Mexico

Authors and Affiliations

  1. Departamento de Neurobiología Celular y Molecular, Laboratorio de Neurobiología Molecular y Celular, Instituto de Neurobiología, Campus UNAM Juriquilla, Querétaro, Qro, CP 76230, Mexico

    L. D. Ochoa-de la Paz, D. B. Salazar-Soto, J. P. Reyes, R. Miledi & A. Martinez-Torres

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  1. L. D. Ochoa-de la Paz
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  2. D. B. Salazar-Soto
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Correspondence to A. Martinez-Torres.

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Ochoa-de la Paz, L.D., Salazar-Soto, D.B., Reyes, J.P. et al. A hyperpolarization-activated ion current of amphibian oocytes. Pflugers Arch - Eur J Physiol 465, 1087–1099 (2013). https://doi.org/10.1007/s00424-013-1231-2

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  • DOI: https://doi.org/10.1007/s00424-013-1231-2

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