Role of Ca2+ fluctuations in L6 myotubes in the regulation of the hexokinase II gene

Expression of the hexokinase (HK) II gene in skeletal muscle is upregulated by electrically stimulated muscle contraction and moderate-intensity exerc ise. However, the molecular mechanism by which this occurs is unknown. Alte rations in intracellular Ca2+ homeostasis accompany contraction and regulat e gene expression in contracting skeletal muscle. Therefore, as a first ste p in understanding the exercise-induced increase in HK II, the ability of C a2+ to increase HK II mRNA was investigated in cultured skeletal muscle cel ls, namely L6 myotubes. Exposure of cells to the ionophore A-23187 resulted in an approximately threefold increase in HK II mRNA. Treatment of cells w ith the extracellular Ca2+ chelator EGTA did not alter HK II mRNA, nor was it able to prevent the A-23187-induced increase. Treatment of cells with th e intracellular Ca2+ chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetra acetic acid tetra(acetoxymethyl) ester (BAPTA-AM) also resulted in an appro ximately threefold increase in HK II mRNA in the absence of ionophore, whic h was similar to the increase in HK II mRNA induced by the combination of B APTA-AM and A-23187. In summary, a rise in intracellular Ca2+ is not necess ary for the A-23187-induced increase in HK II mRNA, and increases in HK II mRNA occur in response to treatments that decrease intracellular Ca2+ store s. Depletion of intracellular Ca2+ stores may be one mechanism by which mus cle contraction increases HK II mRNA.