Generation of immortalized murine forebrain cell lines expressing an alpha isoform of Ca2+/calmodulin-dependent protein kinase II.
Immortalized hybrid cells were generated by the somatic fusion of the cells from the forebrain of embryonic mouse with N18TG2 neuroblastoma cells. Three monoclonal hybrid cell lines, designated NF26, NF81, and NF83 (neuroblastoma forebrain hybrid cells), expressing an alpha isoform of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) were isolated, and their expression was demonstrated by immunoblotting and immunocytochemistry using a monoclonal antibody specific to the a isoform of the enzyme. The kinase activity of the hybrid cells was 2- to 3-fold higher than that of the parent neuroblastoma line N18TG2 cells. The neuronal origin of these lines was shown by their immunoreactivity to neurofilament protein, a neuron specific marker. Lines NF26, NF81, and NF83 are the first cell lines to express the gene of the alpha isoform of CaM kinase II in the brain.
Fujii
M
M
Department of Biochemistry, Faculty of Pharmaceutical Sciences, University of Tokushima, Shomachi, Japan.
Nomura
T
T
Kumatoriya
K
K
Yoshimura
Y
Y
Yamauchi
T
T
D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism; D002999:Clone Cells; D015971:Gene Expression Regulation, Enzymologic; D006822:Hybrid Cells; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism*; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000201:enzymology; D016548:Prosencephalon / Q000166:cytology / Q000201:enzymology*; D051381:Rats; D014407:Tumor Cells, Cultured
D007527:Isoenzymes; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases