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Using SILAC to Study Cell Signaling in Neurons

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Air date: Friday, December 7, 2007, 10:00:00 AM
Time displayed is Eastern Time, Washington DC Local
Views: Total views: 43 * This only includes stats from October 2011 and forward.
Category: Proteomics
Runtime: 01:05:58
Description: The formation and refinement of connections between neurons in the developing brain, and modulation of synaptic strength in the adult brain often rely on the stimulation of receptor tyrosine kinases by their ligands. Much can be learned about how this signaling works by using stable isotope labeling quantitative proteomic methods such as Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and targeted protein isolation strategies. These sample preparation methods are then combined with liquid chromatography-Q-TOF or LTQ-Orbitrap tandem mass spectrometry to identify and compare the relative amounts of proteins in signal transduction complexes from stimulated and nonstimulated cells. I will describe the use of SILAC in our lab to study ephrin signaling in NG108 cell cultures and BDNF signaling in primary cortical neurons. I will also describe SILAC experiments to characterize activity-dependent changes in postsynaptic density composition in primary neuronal cell culture.

http://proteome.nih.gov
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NLM Title: Using SILAC to study cell signaling in neurons [electronic resource] / Thomas A. Neubert.
Author: Neubert, Thomas A.
National Institutes of Health (U.S.). Proteomics Interest Group.
Publisher:
Abstract: (CIT): The formation and refinement of connections between neurons in the developing brain, and modulation of synaptic strength in the adult brain often rely on the stimulation of receptor tyrosine kinases by their ligands. Much can be learned about how this sig. The formation and refinement of connections between neurons in the developing brain, and modulation of synaptic strength in the adult brain often rely on the stimulation of receptor tyrosine kinases by their ligands. Much can be learned about how this signaling works by using stable isotope labeling quantitative proteomic methods such as Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and targeted protein isolation strategies. These sample preparation methods are then combined with liquid chromatography-Q-TOF or LTQ-Orbitrap tandem mass spectrometry to identify and compare the relative amounts of proteins in signal transduction complexes from stimulated and nonstimulated cells. Dr. Neubert will describe the use of SILAC in our lab to study ephrin signaling in NG108 cell cultures and BDNF signaling in primary cortical neurons and also describe SILAC experiments to characterize activity-dependent changes in postsynaptic density composition in primary neuronal cell culture.
Subjects: Amino Acids--metabolism
Isotope Labeling
Neurons--physiology
Signal Transduction--physiology
Publication Types: Webcasts
Rights: This is a work of the United States Government.
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NLM Classification: WL 102.5
NLM ID: 101463202
CIT Live ID: 6276
Permanent link: https://videocast.nih.gov/launch.asp?14188