PMID- 18256212
OWN - NLM
STAT- MEDLINE
DCOM- 20080821
LR  - 20211020
IS  - 1535-9484 (Electronic)
IS  - 1535-9476 (Print)
IS  - 1535-9476 (Linking)
VI  - 7
IP  - 6
DP  - 2008 Jun
TI  - Stable isotopic labeling by amino acids in cultured primary neurons: application 
      to brain-derived neurotrophic factor-dependent phosphotyrosine-associated 
      signaling.
PG  - 1067-76
LID - 10.1074/mcp.M700387-MCP200 [doi]
AB  - Cultured primary neurons are a well established model for the study of neuronal 
      function in vitro. Here we demonstrated that stable isotope labeling by amino 
      acids in cell culture (SILAC) can be applied to a differentiated, non-dividing 
      cell type such as primary neurons, and we applied this technique to assess 
      changes in the neuronal phosphotyrosine proteome in response to stimulation by 
      brain-derived neurotrophic factor (BDNF), an important molecule for the 
      development and regulation of neuronal connections. We found that 13 proteins had 
      SILAC ratios above 1.50 or below 0.67 in phosphotyrosine immunoprecipitations 
      comparing BDNF-treated and control samples, and an additional 18 proteins had 
      ratios above 1.25 or below 0.80. These proteins include TrkB, the receptor 
      tyrosine kinase for BDNF, and others such as hepatocyte growth factor-regulated 
      tyrosine kinase substrate and signal-transducing adaptor molecule, which are 
      proteins known to regulate intracellular trafficking of receptor tyrosine 
      kinases. These results demonstrate that the combination of primary neuronal cell 
      culture and SILAC can be a powerful tool for the study of the proteomes of 
      neuronal molecular and cellular dynamics.
FAU - Spellman, Daniel S
AU  - Spellman DS
AD  - Department of Pharmacology, Skirball Institute of Biomolecular Medicine, New York 
      University School of Medicine, New York, New York 10016, USA.
FAU - Deinhardt, Katrin
AU  - Deinhardt K
FAU - Darie, Costel C
AU  - Darie CC
FAU - Chao, Moses V
AU  - Chao MV
FAU - Neubert, Thomas A
AU  - Neubert TA
LA  - eng
GR  - P01 HD023315/HD/NICHD NIH HHS/United States
GR  - RR017990/RR/NCRR NIH HHS/United States
GR  - NS21072/NS/NINDS NIH HHS/United States
GR  - R01 NS021072/NS/NINDS NIH HHS/United States
GR  - HD23315/HD/NICHD NIH HHS/United States
GR  - P30 NS050276/NS/NINDS NIH HHS/United States
GR  - R56 NS021072/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20080206
PL  - United States
TA  - Mol Cell Proteomics
JT  - Molecular & cellular proteomics : MCP
JID - 101125647
RN  - 0 (Amino Acids)
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Peptides)
RN  - 21820-51-9 (Phosphotyrosine)
RN  - EC 2.7.10.1 (Receptor, trkB)
SB  - IM
MH  - Amino Acids/*chemistry
MH  - Animals
MH  - Brain-Derived Neurotrophic Factor/*metabolism
MH  - Cell Culture Techniques
MH  - Cells, Cultured
MH  - Cerebral Cortex/metabolism
MH  - Hippocampus/metabolism
MH  - Models, Biological
MH  - Neurons/*cytology/metabolism
MH  - Peptides/chemistry
MH  - Phosphotyrosine/*chemistry
MH  - Protein Transport
MH  - Rats
MH  - Receptor, trkB/metabolism
MH  - Signal Transduction
PMC - PMC2424194
EDAT- 2008/02/08 09:00
MHDA- 2008/08/22 09:00
PMCR- 2009/06/01
CRDT- 2008/02/08 09:00
PHST- 2008/02/08 09:00 [pubmed]
PHST- 2008/08/22 09:00 [medline]
PHST- 2008/02/08 09:00 [entrez]
PHST- 2009/06/01 00:00 [pmc-release]
AID - S1535-9476(20)31349-9 [pii]
AID - M700387-MCP200 [pii]
AID - 10.1074/mcp.M700387-MCP200 [doi]
PST - ppublish
SO  - Mol Cell Proteomics. 2008 Jun;7(6):1067-76. doi: 10.1074/mcp.M700387-MCP200. Epub 
      2008 Feb 6.