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Truncation of murine CaV1.2 at Asp 1904 increases CaV1.3 expression in embryonic atrial cardiomyocytes

  • Ion channels, Receptors and Transporters
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Abstract

Cardiac CaV1.2 channels play a critical role in cardiac function. It has been proposed that the carboxyl-terminal intracellular tail of the CaV1.2 channel is the target of Ca2+-dependent and Ca2+-independent regulation of the channel. Recent studies on C-terminal truncated forms of the CaV1.2 channel reported neonatal death, reduced CaV1.2 current, and failure of β-adrenergic stimulation of these channels in ventricular cardiomyocytes (CMs). Here, we used atrial CMs at embryonic day 18.5 that expressed a C-terminal truncated form of the CaV1.2 channel (Stop/Stop). Surprisingly, the atrial CMs showed robust L-type Ca2+ currents which could be stimulated by forskolin, an activator of adenylyl cyclase. These currents exhibited a left-ward shift in the voltage-dependent activation curve and a reduced sensitivity to the Ca2+ channel blocker isradipine as compared to currents in wild-type atrial CMs. RT-PCR analysis revealed normal levels of mRNA for the CaV1.2 channel but a twofold increase in the level of mRNA for the CaV1.3 channel in the Stop/Stop atrium as compared to wild-type atrium. A Western blot analysis indicated an increase of CaV1.3 protein in the Stop/Stop atrium. We suggest that, in contrast to Stop/Stop ventricular CMs, Stop/Stop atrial CMs can compensate the functional loss of the truncated CaV1.2 channel with an upregulation of the CaV1.3 channel.

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Abbreviations

CMs:

Cardiomyocytes

GAPDH:

Glycerinaldehyd-3-phosphat-dehydrogenase

HPRT:

Hypoxanthine-guanine phosphoribosyltransferase

E18.5:

Embryonic day 18.5

AKAP:

A kinase anchoring protein

PKA:

cAMP-dependent protein kinase

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Authors and Affiliations

  1. Forschergruppe 923, Institut für Pharmakologie und Toxikologie, TU München, Biedersteiner Str. 29, 80802, München, Germany

    Jie Ding, Katrin Domes, Franz Hofmann & Jörg W Wegener

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  1. Jie Ding
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  2. Katrin Domes
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  3. Franz Hofmann
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  4. Jörg W Wegener
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Correspondence to Jörg W Wegener.

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Authors Jie Ding and Katrin Domes contributed equally to the manuscript.

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Supplemental Fig. 1

Expression analysis of L-type Ca2+ channels in the ventricular and atrial tissue from WT and Stop/Stop mice by RT-PCR. Representative ethidium bromide-stained agarose gel of amplicons generated with primers against mRNA of CaV1.2b (a) and CaV2.1 (b) in ventricular (V) and atrial (A) tissue from WT and Stop/Stop mice. The size of the amplicons is indicated on the left (base pair). Primers against mRNA for GAPDH or HPRT were used as control. In a, lung tissue from WT mice was used as a control for the smooth muscle specificity of the primers against mRNA of CaV1.2b, and H2O indicates a control for the specificity of the amplification process. c, d Semiquantitative analysis of the amplification products. Columns represent means ± SEM. Numbers in the columns indicate the number of experiments. Statistical analysis was performed by Student t test revealing no statistically significant difference between the data sets (JPEG 69 kb)

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Ding, J., Domes, K., Hofmann, F. et al. Truncation of murine CaV1.2 at Asp 1904 increases CaV1.3 expression in embryonic atrial cardiomyocytes. Pflugers Arch - Eur J Physiol 465, 955–964 (2013). https://doi.org/10.1007/s00424-012-1212-x

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