Exp Mol Med.  2009 Aug;41(8):601-610. 10.3858/emm.2009.41.8.066.

Neuronal activation increases the density of eukaryotic translation initiation factor 4E mRNA clusters in dendrites of cultured hippocampal neurons

Affiliations
  • 1Department of Anatomy, College of Medicine, Dongguk University, Gyeongju 780-714, Korea. moonis@dongguk.ac.kr
  • 2Department of Biochemistry, College of Medicine, Inje University, Pusan 614-735, Korea.
  • 3Department of Physiology and Neurobiology, University of Connecticut, CT 06269, USA.

Abstract

Activity-dependent dendritic translation in CNS neurons is important for the synapse-specific provision of proteins that may be necessary for strengthening of synaptic connections. A major rate-limiting factor during protein synthesis is the availability of eukaryotic translation initiation factor 4E (eIF4E), an mRNA 5'-cap-binding protein. In this study we show by fluorescence in situ hybridization (FISH) that the mRNA for eIF4E is present in the dendrites of cultured rat hippocampal neurons. Under basal culture conditions, 58.7 +/- 11.6% of the eIF4E mRNA clusters localize with or immediately adjacent to PSD-95 clusters. Neuronal activation with KCl (60 mM, 10 min) very significantly increases the number of eIF4E mRNA clusters in dendrites by 50.1 and 74.5% at 2 and 6 h after treatment, respectively. In addition, the proportion of eIF4E mRNA clusters that localize with PSD-95 increases to 74.4 +/- 7.7% and 77.8 +/- 7.6% of the eIF4E clusters at 2 and 6 h after KCl treatment, respectively. Our results demonstrate the presence of eIF4E mRNA in dendrites and an activity-dependent increase of these clusters at synaptic sites. This provides a potential mechanism by which protein translation at synapses may be enhanced in response to synaptic stimulation.

Keyword

dendrites; eukaryotic initiation factor-4E; immunohistochemistry; in situ hybridization, fluorescence; potassium chloride; protein biosynthesis; synapses

MeSH Terms

Animals
Cells, Cultured
Dendrites/*metabolism
Eukaryotic Initiation Factor-4E/genetics/*metabolism
Hippocampus/*cytology
Immunohistochemistry
In Situ Hybridization, Fluorescence
Microscopy, Confocal
Neurons/cytology/*physiology
Potassium Chloride/pharmacology
Protein Biosynthesis
RNA, Messenger/genetics/*metabolism
Rats
Rats, Sprague-Dawley
Synapses
*Up-Regulation
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