PMID- 10800934 OWN - NLM STAT- MEDLINE DCOM- 20000525 LR - 20190630 IS - 0022-3042 (Print) IS - 0022-3042 (Linking) VI - 74 IP - 5 DP - 2000 May TI - Increase of brain-derived neurotrophic factor gene expression in NG108-15 cells by the nuclear isoforms of Ca2+/ calmodulin-dependent protein kinase II. PG - 1913-22 AB - We have reported that the delta3 isoform of Ca2+/ calmodulin-dependent protein kinase II (CaM kinase II) is abundant in the nucleus in cerebellar granule cells. To examine the possibility that the nuclear isoforms of CaM kinase II are involved in the expression of brain-derived neurotrophic factor (BDNF), we transiently overexpressed the delta3 isoform in NG108-15 cells. The quantitative RT-PCR analysis revealed that rat cerebellum and NG108-15 cells expressed the exon IV-containing mRNA of BDNF (exon IV-BDNF mRNA) more than the exon III-BDNF mRNA. Treatment of NG108-15 cells with Bay K 8644 increased both exon III- and exon IV-BDNF mRNAs, and overexpression of the 83 isoform potentiated the expression of the exon IV-BDNF mRNA. The potentiation was not observed in the cells that were overexpressed with either the 61 isoform, a nonnuclear isoform, or the inactive mutant of the delta3 isoform. We constructed the luciferase reporter gene following the promoter upstream of exon IV and confirmed that overexpression of the delta3 isoform increased luciferase gene expression. Double-immunostaining of NG108-15 cells with the antibodies to CaM kinase II and BDNF clearly showed that BDNF was highly expressed in the cells that were overexpressed with the delta3 isoform or the alphaB isoform, another nuclear isoform of CaM kinase II. These results suggest that the nuclear isoforms of CaM kinase II are involved in the expression of BDNF. FAU - Takeuchi, Y AU - Takeuchi Y AD - Department of Pharmacology, Kumamoto University School of Medicine, Japan. FAU - Yamamoto, H AU - Yamamoto H FAU - Miyakawa, T AU - Miyakawa T FAU - Miyamoto, E AU - Miyamoto E LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - England TA - J Neurochem JT - Journal of neurochemistry JID - 2985190R RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (RNA, Messenger) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 2) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinases) SB - IM MH - Animals MH - Brain-Derived Neurotrophic Factor/*genetics/metabolism MH - Calcium-Calmodulin-Dependent Protein Kinase Type 2 MH - Calcium-Calmodulin-Dependent Protein Kinases/genetics/metabolism/*physiology MH - Cell Line MH - Cell Nucleus/*enzymology MH - Cerebellum/metabolism MH - Cerebral Cortex/metabolism MH - *Gene Expression Regulation, Enzymologic MH - Immunologic Techniques MH - RNA, Messenger/metabolism MH - Rats MH - Staining and Labeling MH - Transfection EDAT- 2000/05/09 09:00 MHDA- 2000/06/08 09:00 CRDT- 2000/05/09 09:00 PHST- 2000/05/09 09:00 [pubmed] PHST- 2000/06/08 09:00 [medline] PHST- 2000/05/09 09:00 [entrez] AID - 10.1046/j.1471-4159.2000.0741913.x [doi] PST - ppublish SO - J Neurochem. 2000 May;74(5):1913-22. doi: 10.1046/j.1471-4159.2000.0741913.x.