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Study of protein and RNA in dendritic spines using multi‐isotope imaging mass spectrometry
Author(s) -
Brismar H.,
Aperia A.,
Westin L.,
Moy J.,
Wang M.,
Guillermier C.,
Poczatek J. C.,
Lechene C.
Publication year - 2014
Publication title -
surface and interface analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.52
H-Index - 90
eISSN - 1096-9918
pISSN - 0142-2421
DOI - 10.1002/sia.5617
Subject(s) - dendritic spine , rna , chemistry , microbiology and biotechnology , spine (molecular biology) , protein biosynthesis , organelle , synapse , biology , biophysics , biochemistry , neuroscience , gene , hippocampal formation
The classical view of neuronal protein synthesis is that proteins are made in the cell body and then transported to their functional sites in the dendrites and the dendritic spines. Indirect evidence, however, suggests that protein synthesis can directly occur in the distal dendrites, far from the cell body. We are developing protocols for dual labeling of RNA and proteins using 15 N‐uridine and 18 O‐ or 13 C‐leucine pulse chase in cultured neurons to identify and localize both protein synthesis and fate of newly synthesized proteins. Pilot experiments show discrete localization of both RNA and newly synthesized proteins in dendrites, close to dendritic spines. We have for the first time directly imaged and measured the production of proteins at the subcellular level in the neuronal dendrites, close to the functional sites, the dendritic spines. This will open a powerful way to study neural growth and synapse plasticity in health and disease. Copyright © 2014 John Wiley & Sons, Ltd.