Abstract
The sphingosine kinase inhibitor, dimethylsphingosine, is an important tool for investigating intracellular effects of the putative second messenger compound, sphingosine 1-phosphate. However, the specificity of action of dimethylsphingosine has not been fully investigated. In human SH-SY5Y neuroblastoma cells, dimethylsphingosine (30 μM), produced a 25-fold increase in the EC 50 for methacholine-induced Ca 2+ mobilisation, and reduced the maximum response by 57±5%, suggesting the involvement of sphingosine 1-phosphate production in the Ca 2+ signal. However, dimethylsphingosine also inhibited [ 3H] N-methylscopolamine binding to whole SH-SY5Y cells and reduced methacholine-induced phosphoinositide turnover. Thus, this compound must be used with caution when investigating the role of sphingosine kinase in G-protein coupled receptor-mediated Ca 2+ mobilisation responses.
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