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Expression of the muscular dystrophy-associated caveolin-3P104L mutant in adult mouse skeletal muscle specifically alters the Ca2+ channel function of the dihydropyridine receptor

  • Ion Channels, Receptors and Transporters
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Abstract

Caveolins are plasma-membrane-associated proteins potentially involved in a variety of signalling pathways. Different mutations in CAV3, the gene encoding for the muscle-specific isoform caveolin-3 (Cav-3), lead to muscle diseases, but the underlying molecular mechanisms remain largely unknown. Here, we explored the functional consequences of a Cav-3 mutation (P104L) inducing the 1C type limb-girdle muscular dystrophy (LGMD 1C) in human on intracellular Ca2+ regulation of adult skeletal muscle fibres. A YFP-tagged human Cav-3P104L mutant was expressed in vivo in muscle fibres from mouse. Western blot analysis revealed that expression of this mutant led to an ∼80% drop of the level of endogenous Cav-3. The L-type Ca2+ current density was found largely reduced in fibres expressing the Cav-3P104L mutant, with no change in the voltage dependence of activation and inactivation. Interestingly, the maximal density of intramembrane charge movement was unaltered in the Cav-3P104L-expressing fibres, suggesting no change in the total amount of functional voltage-sensing dihydropyridine receptors (DHPRs). Also, there was no obvious alteration in the properties of voltage-activated Ca2+ transients in the Cav-3P104L-expressing fibres. Although the actual role of the Ca2+ channel function of the DHPR is not clearly established in adult skeletal muscle, its specific alteration by the Cav-3P104L mutant suggests that it may be involved in the physiopathology of LGMD 1C.

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Abbreviations

Cav-3:

Caveoline-3

DHPR:

Dihydropyridines receptor

RyR:

Ryanodine receptor

FDB:

Flexor digitorum brevis

YFP:

Yellow fluorescent protein

VGCC:

Voltage-gated calcium channel

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Acknowledgements

We are very grateful to Dr. R.G. Parton (University of Queensland, Australia) for the gift of the wild-type, P104L and C71W caveolin cDNAs. This work was supported by grants from the Centre National de la Recherche Scientifique (CNRS), Agence Nationale de la Recherche (#05-MRAR-001-01), University Lyon 1 and Association Française contre les Myopathies.

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Authors and Affiliations

  1. Physiologie Intégrative Cellulaire et Moléculaire, Université Claude Bernard—Lyon 1, UMR CNRS 5123, Bât. Raphaël Dubois, 43 boulevard du 11 novembre 1918, 69622, Villeurbanne Cedex, France

    Norbert Weiss, Harold Couchoux, Claude Legrand, Christine Berthier, Bruno Allard & Vincent Jacquemond

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  1. Norbert Weiss
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  2. Harold Couchoux
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  3. Claude Legrand
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  4. Christine Berthier
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Correspondence to Vincent Jacquemond.

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Fig. S1

Over-expression of Cav-3P104L-YFP does not alter action potential properties of the skeletal muscle fibre. a Representative records of action potentials from a control fibre (top panel) and from a Cav-3P104L-YFP-expressing fibre (bottom panel). b–d Corresponding mean values for APs amplitude, maximum rate of rise (dV/dt)max and duration measured at 25%, 50% and 75% of repolarisation. There is no significant difference between APs recorded from control (n = 9) and Cav-3P104L-YFP-expressing fibres (n = 11) (GIF 231 KB).

Fig. S1

High resolution image file (TIFF 3.48 MB).

Fig. S2

Indo-1 calcium transients elicited by depolarisation to +10 mV of increasing duration in control and Cav-3C71W-YFP-expressing fibres. a Indo-1 saturation signals in response to successive depolarisations of 5-, 10- and 20-ms duration to +10 mV from a holding potential of −80 mV in a control (left panel) and in Cav-3C71W-YFP-expressing fibre (right panel). b–e Corresponding mean values for the initial resting [Ca2+], peak change in [Ca2+], time constant τ of [Ca2+] decay and final [Ca2+] level, respectively. No significant difference in any parameter was detected between control and Cav-3C71W-expressing fibres (GIF 129 KB).

Fig. S2

High resolution image file (TIFF 4.20 MB).

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Weiss, N., Couchoux, H., Legrand, C. et al. Expression of the muscular dystrophy-associated caveolin-3P104L mutant in adult mouse skeletal muscle specifically alters the Ca2+ channel function of the dihydropyridine receptor. Pflugers Arch - Eur J Physiol 457, 361–375 (2008). https://doi.org/10.1007/s00424-008-0528-z

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