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Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
Published on: June 17, 2015
The mRNA for elongation factor 1alpha is localized in dendrites and translated in response to treatments that induce
Fen Huang1, Jennifer K Chotiner, Oswald Steward
1Reeve-Irvine Research Center, Department of Anatomy and Neurobiology, University of California, Irvine, California 92697, USA.
Abstract:
There is increasing evidence that long-lasting forms of activity-dependent synaptic plasticity, such as long-term potentiation (LTP) and long-term depression (LTD), require local synthesis of proteins within dendrites. Identifying novel dendritic mRNAs and determining how their distribution and translation is regulated is a high priority. We demonstrate here that the mRNA for the elongation factor 1 alpha (EF1alpha) is present in vivo in the dendrites of neurons that exhibit LTP and LTD, and that its translation is locally regulated. The subcellular distribution of EF1alpha mRNA differs from any of the dendritic mRNAs that have been described previously. In the hippocampus, the mRNA is highly expressed in cell bodies and is also concentrated in the zone of termination of commissural/associational afferents in the inner molecular layer, suggesting that mRNA localization is in some way related to the distribution of different types of synapses. Nevertheless, the localization of EF1alpha mRNA is not altered by prolonged periods of synaptic activation that are sufficient to cause a dramatic redistribution of Arc mRNA. Local application of the metabotropic glutamate receptor agonist (R,S)-3,5-dihydroxyphenylglycine (DHPG) led to dramatic increases in immunostaining for EF1alpha protein in dendrites, and treatment of hippocampal slices with DHPG, which is known to induce LTD, led to increases in EF1alpha protein levels. Both responses were blocked by the protein synthesis inhibitor anisomycin. In contrast, stimulation of the perforant path using patterns of stimulation that induce LTP caused rapid increases of immunostaining for EF1alpha protein in the activated dendritic lamina, but these increases were not blocked by anisomycin or rapamycin. The findings suggest that local synthesis of EF1alpha protein may be important for the synaptic mechanisms that underlie protein synthesis-dependent LTD.
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