Exposure of C2C12 muscle cells to hypertonic stress induced an increase in cell content of creatine transporter mRNA and of creatine transport activity, which peaked after about 24 h incubation at 0.45 osmol (kg H(2)O)(-1). This induction of transport activity was prevented by addition of either cycloheximide, to inhibit protein synthesis, or of actinomycin D, to inhibit RNA synthesis. Creatine uptake by these cells is largely Na(+) dependent and kinetic analysis revealed that its increase under hypertonic conditions resulted from an increase in V(max) of the Na(+)-dependent component, with no significant change in the K(m) value of about 75 mumol l(-1). Quantitative real-time PCR revealed a more than threefold increase in the expression of creatine transporter mRNA in cells exposed to hypertonicity. Creatine supplementation significantly enhanced survival of C2C12 cells incubated under hypertonic conditions and its effect was similar to that obtained with the well known compatible osmolytes, betaine, taurine and myo-inositol. This effect seemed not to be linked to the energy status of the C2C12 cells because hypertonic incubation caused a decrease in their ATP content, with or without the addition of creatine at 20 mmol l(-1) to the medium. This induction of creatine transport activity by hypertonicity is not confined to muscle cells: a similar induction was shown in porcine endothelial cells.
Creatine as a compatible osmolyte in muscle cells exposed to hypertonic stress / Alfieri, R; Bonelli, M; Cavazzoni, A; Brigotti, Maurizio; Fumarola, C; Sestili, Piero; Mozzoni, P; DE PALMA, Giuseppe; Mutti, A; Carnicelli, D; Vacondio, F; Silva, C; Borghetti, A. F; Wheeler, K. P; Petronini, P.. - In: THE JOURNAL OF PHYSIOLOGY. - ISSN 0022-3751. - 576:(2006), pp. 391-401. [10.1113/jphysiol.2006.115006]
Creatine as a compatible osmolyte in muscle cells exposed to hypertonic stress
Alfieri, R;Bonelli, M;Cavazzoni, A;BRIGOTTI, MAURIZIO;Fumarola, C;SESTILI, PIERO;Mozzoni, P;DE PALMA, Giuseppe;Mutti, A;Vacondio, F;Silva, C;Borghetti, A. F;Wheeler, K. P;Petronini, P.
2006-01-01
Abstract
Exposure of C2C12 muscle cells to hypertonic stress induced an increase in cell content of creatine transporter mRNA and of creatine transport activity, which peaked after about 24 h incubation at 0.45 osmol (kg H(2)O)(-1). This induction of transport activity was prevented by addition of either cycloheximide, to inhibit protein synthesis, or of actinomycin D, to inhibit RNA synthesis. Creatine uptake by these cells is largely Na(+) dependent and kinetic analysis revealed that its increase under hypertonic conditions resulted from an increase in V(max) of the Na(+)-dependent component, with no significant change in the K(m) value of about 75 mumol l(-1). Quantitative real-time PCR revealed a more than threefold increase in the expression of creatine transporter mRNA in cells exposed to hypertonicity. Creatine supplementation significantly enhanced survival of C2C12 cells incubated under hypertonic conditions and its effect was similar to that obtained with the well known compatible osmolytes, betaine, taurine and myo-inositol. This effect seemed not to be linked to the energy status of the C2C12 cells because hypertonic incubation caused a decrease in their ATP content, with or without the addition of creatine at 20 mmol l(-1) to the medium. This induction of creatine transport activity by hypertonicity is not confined to muscle cells: a similar induction was shown in porcine endothelial cells.File | Dimensione | Formato | |
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