© 2001 by European Society of Cardiology
Copyright © 2000, European Society of Cardiology
Transient down-regulation of L-type Ca2+ channel and dystrophin expression after balloon injury in rat aortic cells
aIGH, CNRS UPR 1142, 141 rue de la Cardonille, 34396 Montpellier Cedex 5, France
bCRBM, CNRS UPR 1086, Université de Montpellier I, 1919 route de Mende, 34293 Montpellier Cedex 5, France
cUniversité de Paris VI, service de médecine interne, l'Hôtel-Dieu, 75181 Paris Cedex 04, France
dINSERM U 128, IFR 24, 1919 route de Mende, 34293 Montpellier Cedex 5, France
* Corresponding author. Correspondence address: INSERM U-390, Hôpital Arnaud de Villeneuve, F-34295 Montpellier Cedex 5, France. Tel.: +33-467-415-244; fax: +33-467-415-242 srichard{at}montp.inserm.fr
Objective: Migration and proliferation of arterial smooth muscle cells are critical responses during restenosis after balloon angioplasty. We investigated the changes in the expression of Ca2+ channels and dystrophin, two determinants of contraction, after balloon injury of rat aortas. Methods: Proliferation and migration of aortic myocytes were triggered in vivo by the passage of an inflated balloon catheter in the aortas of 12-week-old male Wistar rats. We used the whole-cell patch clamp technique to investigate Ba2+ currents (IBa) through Ca2+ channels in single cells freshly isolated from media and neointima at various times after injury (days 2, 7, 15, 30 and 45). Results: No T-type Ca2+ channel current was recorded in any cell at any time. In contrast, a dihydropyridine (DHP)-sensitive L-type IBawas recorded consistently in the media of intact aorta. After aortic injury, IBa decreased dramatically (at days 2 and 7) but recovered over time to reach normal amplitude on days 30 and 45. In the neointima, IBa was absent on day 15 but also increased gradually over time as observed at days 30 and 45. The use of a specific antibody directed against the L-type Ca2+ channel 
1C subunit showed, both by immunostaining and by Western blotting, no expression of the Ca2+ channel protein on day 15. Parallel immunodetection of dystrophin showed that this marker of the contractile phenotype of SMCs was also not detectable at this stage in neointimal cells. Both proteins were re-expressed at days 45 and 63. Balloon injury induces a transient down-regulation of IBa in arterial cells. Conclusions: Cell dedifferentiation and proliferation in vivo abolish the expression of L-type Ca2+ channels and dystrophin in neointimal cells. These changes may be critical in the regulation of Ca2+ homeostasis and, thereby, contraction of the arterial SMCs during restenosis following angioplasty.
KEYWORDS Ca2+: calcium; [Ca2+]i, cytosolic free calcium concentration; DHP, dihydropyridine; SMCs, smooth muscle cells; FITC, fluorescein isothiocyanate; SDS, sodium dodecyl sulfate; HEPES, N-2-hydroxyethylpiperazine-N'-2-ethane sulfonic acid h-CaD: heavy-molecular-weight caldesmon; l-CaD, low-molecular-weight caldesmon; PMSF, phenyl methyl sulfonyl fluoride; CsOH, caesium hydroxide; TBS–T, Tris–HCl buffered saline–Tween; EGTA, ethylene glycol-bis (β-aminoethylether)N,N,N',N'-tetraacetic acid; BSA, bovine serum albumin; ATP, adenosine-5'-triphosphate; GTP, guanosine-5'-triphosphate
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