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J. Biol. Chem., Vol. 278, Issue 25, 22316-22324, June 20, 2003
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Induction of T-type Calcium Channel Gene Expression by Chronic Hypoxia*
From the Laboratorio de Investigaciones Biomédicas, Departamento de Fisiología and Hospital Universitario Virgen del Rocío, Universidad de Sevilla, E-41013 Seville, Spain
Cellular responses to hypoxia can be acute or chronic. Acute responses mainly depend on oxygen-sensitive ion channels, whereas chronic responses rely on the hypoxia-inducible transcription factors (HIFs), which up-regulate the expression of enzymes, transporters, and growth factors. It is unknown whether the expression of genes coding for ion channels is also influenced by hypoxia. We report here that the 
1H gene of T-type voltage-gated calcium channels is highly induced by lowering oxygen tension in PC12 cells. Accumulation of 1H mRNA in response to hypoxia is time- and dose-dependent and paralleled by an increase in the density of T-type calcium channel current recorded in patch clamped cells. HIF appears to be involved in the response to hypoxia, since cobalt chloride, desferrioxamine, and dimethyloxalylglycine, compounds that mimic HIF-regulated gene expression, replicate the hypoxic effect. Moreover, functional inhibition of HIF-2 protein accumulation using antisense HIF-2 oligonucleotides reverses the effect of hypoxia on T-type Ca2+ channel expression. Importantly, regulation by oxygen tension is specific for T-type calcium channels, since it is not observed with the L-, N-, and P/Q-channel types. These findings show for the first time that hypoxia induces an ion channel gene via a HIF-dependent mechanism and define a new role for the T-type calcium channels as regulators of cellular excitability and calcium influx under chronic hypoxia.
Received for publication, December 10, 2002 , and in revised form, April 4, 2003.
* This work was supported by grants from Fondo de Investigación Sanitaria, the Andalusian Government, and the Ramón Areces Foundation (to M. D. C. and J. L.-B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
These authors contributed equally to this work.
¶ Present address: Servicio de Otorrinolaringología, Hospital Universitario Central de Asturias, C/Celestino Villamil s/n E-33006, Oviedo, Spain.
Recipient of the "Ayuda a la investigación 2000" of the Juan March Foundation. To whom correspondence should be addressed: Laboratorio de Investigaciones Biomédicas, Edificio de Laboratorios, 2a Planta, Hospital Universitario Virgen del Rocío, Avenida Manuel Siurot s/n, E-41013, Seville, Spain. Tel.: 34-955-012648 or 34-955-013157; Fax: 34-954-617301; E-mail: lbarneo{at}us.es.
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