Bruton's tyrosine kinase activity and inositol 1,4,5-trisphosphate production are not altered in DT40 lymphoma B cells exposed to power line frequency magnetic fields.
Exposure of wild-type DT40 lymphoma B cells or Bruton's tyrosine kinase (BTK)-deficient DT40 cells reconstituted with the human btk gene to a 1-gauss 60-Hz electromagnetic field (EMF) has been reported to rapidly increase inositol 1,4,5-trisphosphate (Ins 1,4, 5-P3) production (1,2). Here we have used BTK-deficient DT40 B cells reconstituted with the human btk gene to evaluate the reproducibility of these findings. An experimental design with blinded exposures and anti-IgM treatment to induce Ins 1,4,5-P3 production as a positive control, showed no significant effect of a 1-gauss 60-Hz EMF on Ins 1,4,5-P3 production. Because recent work has shown that the activation of BTK was required for EMF-responsiveness (2), we also evaluated the reproducibility of this finding in wild-type DT40 cells. BTK was activated in a dose- and time-dependent manner by treatment with the tyrosine phosphatase inhibitor pervanadate. However, the ability to detect BTK activation, as measured by increased autophosphorylation by immune complex kinase assay, was dependent on the kinase buffer. Using cells from the original investigators, no evidence was obtained to support the hypothesis that exposure to a 1-gauss 60-Hz EMF had a causal effect on protein-tyrosine kinase activities affecting Ins 1,4,5-P3 production.
Miller
S C
SC
Signal Transduction Laboratory, Pharmaceutical Discovery Division, SRI International, Menlo Park, California 94025, USA. smiller@unix.sri.com
Furniss
M J
MJ
D000077329:Agammaglobulinaemia Tyrosine Kinase; D000906:Antibodies / Q000276:immunology; D004574:Electromagnetic Fields; D004789:Enzyme Activation; D004790:Enzyme Induction; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis*; D016393:Lymphoma, B-Cell / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014638:Vanadates / Q000494:pharmacology
D000906:Antibodies; D007075:Immunoglobulin M; C076715:pervanadate; D014638:Vanadates; D015544:Inositol 1,4,5-Trisphosphate; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase; C000625949:BTK protein, human