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The Journal of Neuroscience, June 1, 2001, 21(11):3797-3805
NMDA Receptor Stimulation and Brain-Derived Neurotrophic Factor Upregulate Homer 1a mRNA via the Mitogen-Activated Protein Kinase Cascade in Cultured Cerebellar Granule Cells
Masaaki Sato, Kazunori Suzuki, and Shigetada Nakanishi Department of Biological Sciences, Faculty of Medicine and Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501, Japan
In three alternative splice variants of Homer 1 transcripts, Homer 1a mRNA has been shown to be upregulated selectively and rapidly by neural stimulation and represents a member of the immediate early gene (IEG) family. We investigated the mechanism underlying Homer 1a mRNA induction in cerebellar granule cell culture. All Homer 1 variants were expressed in cultured granule cells as analyzed by RNA blotting and immunochemical characterization. Glutamate stimulation of granule cells selectively upregulated Homer 1a mRNA via NMDA receptor-mediated influx of extracellular calcium. The induction of Homer 1a mRNA was much slower (peaked at 4 hr) and sustained longer than that of the typical IEG c-fos mRNA. Actinomycin D and cycloheximide experiments have revealed that, despite the presence of the mRNA-destabilizing AU-rich motif, transcriptional activation is a main determinant for selective Homer 1a mRNA induction. Inhibitor analysis as well as immunochemical characterization has indicated that the MEK (MAPK/ERK kinase)-ERK (extracellular signal-regulated kinase) cascade plays an indispensable role in glutamate-stimulated induction of Homer 1a mRNA. Consistent with this observation, brain-derived neurotrophic factor, which is known to activate the ERK cascade, similarly upregulated Homer 1a mRNA. These results demonstrate that MAPK (mitogen-activated protein kinase) is a key mediator that links distinct extracellular stimuli to the transcriptional activation of Homer 1a mRNA.
Key words: granule cell culture; Homer 1; NMDA receptor; brain-derived neurotrophic factor; calcium signaling; MAP kinase; transcriptional activation
Copyright © 2001 Society for Neuroscience 0270-6474/01/21113797-09$05.00/0
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