Calpastatin overexpression in the skeletal muscle of mice prevents clenbuterol-induced muscle hypertrophy and phenotypic shift.
Accumulating evidence suggests that the calpain/calpastatin system is involved in skeletal muscle remodelling induced by β(2) -adrenoceptor agonist treatment. In addition to other pathways, the Akt/mammalian target of rapamycin (mTOR) pathway, controlling protein synthesis, and the calcium/calmodulin-dependent protein kinase 2 (CamK2) and AMP-activated protein kinase (AMPK) pathways, recently identified as calpain substrates, could be relevant in β(2) -adrenoceptor agonist-induced skeletal muscle remodelling. In the present study we investigated muscle hypertrophy and phenotypic shifts, as well as the molecular response of components of the Akt/mTOR pathway (i.e. Akt, eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), ribosomal protein S6 (rpS6), CamK2 and AMPK), in response to calpastatin overexpression in the skeletal muscle of mice treated with 1 mg/kg per day clenbuterol for 21 days. Using gene electrotransfer of a calpastatin expression vector into the tibialis anterior of adult mice, we found that calpastatin overexpression attenuates muscle hypertrophy and phenotypic shifts induced by clenbuterol treatment. At the molecular level, calpastatin overexpression markedly decreased calpain activity, but was ineffective in altering the phosphorylation of Akt, 4E-BP1 and rpS6. In contrast, calpastatin overexpression increased the protein expression of both total AMPK and total CamK2. In conclusion, the results support the contention that the calpain/calpastatin system plays a crucial role in skeletal muscle hypertrophy and phenotypic shifts under chronic clenbuterol treatment, with AMPK and CamK2 probably playing a minor role. Moreover, the calpastatin-induced inhibition of hypertrophy under clenbuterol treatment was not related to a decreased mTOR-dependent initiation of protein translation.
National Institute for Agronomical Research (INRA), Muscular Dynamic and Metabolism, University of Montpellier, Montpellier, France. aymerichpz@yahoo.fr
Aymeric
A
Douillard
Olivier
O
Galbes
Gwenaelle
G
Begue
Bernadette
B
Rossano
John
J
Levin
Barbara
B
Vernus
Anne
A
Bonnieu
Robin
R
Candau
Guillaume
G
Py
D000818:Animals; D002135:Calcium-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D002154:Calpain / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002417:Cattle; D002976:Clenbuterol / Q000037:antagonists & inhibitors / Q000633:toxicity*; D005786:Gene Expression Regulation; D006984:Hypertrophy / Q000139:chemically induced / Q000378:metabolism / Q000517:prevention & control; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D010641:Phenotype
D002135:Calcium-Binding Proteins; C033668:calpastatin; D002154:Calpain; D002976:Clenbuterol