Quantification of the synaptosomal proteome of the rat cerebellum during post-natal development.
Large-scale proteomic analysis of the mammalian brain has been successfully performed with mass spectrometry techniques, such as Multidimensional Protein Identification Technology (MudPIT), to identify hundreds to thousands of proteins. Strategies to efficiently quantify protein expression levels in the brain in a large-scale fashion, however, are lacking. Here, we demonstrate a novel quantification strategy for brain proteomics called SILAM (Stable Isotope Labeling in Mammals). We utilized a (15)N metabolically labeled rat brain as an internal standard to perform quantitative MudPIT analysis on the synaptosomal fraction of the cerebellum during post-natal development. We quantified the protein expression level of 1138 proteins in four developmental time points, and 196 protein alterations were determined to be statistically significant. Over 50% of the developmental changes observed have been previously reported using other protein quantification techniques, and we also identified proteins as potential novel regulators of neurodevelopment. We report the first large-scale proteomic analysis of synaptic development in the cerebellum, and we demonstrate a useful quantitative strategy for studying animal models of neurological disease.
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Daniel B
DB
McClatchy
Lujian
L
Liao
Sung Kyu
SK
Park
John D
JD
Venable
John R
JR
Yates
D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002531:Cerebellum / Q000737:chemistry* / Q000166:cytology / Q000254:growth & development*; D007553:Isotope Labeling; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009587:Nitrogen Isotopes; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012984:Software; D013574:Synaptosomes / Q000737:chemistry*; D053719:Tandem Mass Spectrometry; D013997:Time Factors
D009419:Nerve Tissue Proteins; D009587:Nitrogen Isotopes; D020543:Proteome