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Region-directed phototransfection reveals the functional significance of a dendritically synthesized transcription factor
Journal article   Peer reviewed

Region-directed phototransfection reveals the functional significance of a dendritically synthesized transcription factor

Lindy E Barrett, Jai-Yoon Sul, Hajime Takano, Elisabeth J Van Bockstaele, Philip G Haydon and James H Eberwine
Nature methods, v 3(6), pp 455-460
01 Jun 2006
PMID: 16721379

Abstract

Active Transport, Cell Nucleus - physiology Animals Apoptosis - physiology Cells, Cultured Dendrites - metabolism Dendrites - ultrastructure ets-Domain Protein Elk-1 - genetics ets-Domain Protein Elk-1 - metabolism Hippocampus - cytology Hippocampus - metabolism Photochemistry - methods Rats RNA, Messenger - administration & dosage RNA, Messenger - genetics Transcription Factors - genetics Transcription Factors - metabolism Transfection - methods
Multiple nuclear transcription factors including E-26-like protein 1 (Elk-1) have been found in neuronal dendrites, yet the functional significance of such localization has not yet been explained. Here we use a focal transfection procedure, 'phototransfection', to introduce Elk1 mRNA into specific regions of live, intact primary rat neurons. Introduction and translation of Elk1 mRNA in dendrites produced cell death, whereas introduction and translation of Elk1 mRNA in cell bodies did not produce cell death. Elk-1 translated in dendrites was transported to the nucleus, and cell death depended upon transcription, supporting the dendritic imprinting hypothesis and highlighting the importance of the dendritic environment on protein function. Our demonstration of the utility of phototransfection for spatially controlled introduction of mRNAs opens the broader opportunity to use this method to introduce selected quantities of small molecules into discrete regions of live cells to assess their biological functions.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biochemical Research Methods
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