Role of Ca(2+) fluctuations in L6 myotubes in the regulation of the hexokinase II gene.

Halseth AE, O'Doherty RM, Printz RL, Bracy DP, Granner DK, Wasserman DH
J Appl Physiol (1985). 2000 88 (2): 669-73

PMID: 10658036 · DOI:10.1152/jappl.2000.88.2.669

Expression of the hexokinase (HK) II gene in skeletal muscle is upregulated by electrically stimulated muscle contraction and moderate-intensity exercise. However, the molecular mechanism by which this occurs is unknown. Alterations in intracellular Ca(2+) homeostasis accompany contraction and regulate gene expression in contracting skeletal muscle. Therefore, as a first step in understanding the exercise-induced increase in HK II, the ability of Ca(2+) to increase HK II mRNA was investigated in cultured skeletal muscle cells, namely L6 myotubes. Exposure of cells to the ionophore A-23187 resulted in an approximately threefold increase in HK II mRNA. Treatment of cells with the extracellular Ca(2+) chelator EGTA did not alter HK II mRNA, nor was it able to prevent the A-23187-induced increase. Treatment of cells with the intracellular Ca(2+) chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA-AM) also resulted in an approximately threefold increase in HK II mRNA in the absence of ionophore, which was similar to the increase in HK II mRNA induced by the combination of BAPTA-AM and A-23187. In summary, a rise in intracellular Ca(2+) is not necessary for the A-23187-induced increase in HK II mRNA, and increases in HK II mRNA occur in response to treatments that decrease intracellular Ca(2+) stores. Depletion of intracellular Ca(2+) stores may be one mechanism by which muscle contraction increases HK II mRNA.

MeSH Terms (13)

Animals Calcimycin Calcium Cell Line Chelating Agents Dose-Response Relationship, Drug Egtazic Acid Extracellular Space Gene Expression Regulation, Enzymologic Hexokinase Isoenzymes Muscle, Skeletal RNA, Messenger

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