Quantitative proteomic analysis of primary neurons reveals diverse changes in synaptic protein content infmr1knockout mice

• DOI: 10.1073/pnas.0804678105
• Publication Date: 2008-10-01T03:04:45Z
• Authors: Lujian Liao; Sung Kyu Park; Tao Xu; Peter Vanderklish; John R. Yates
• Keywords: FMR1; Knockout mouse

Fragile X syndrome (FXS) is a common inherited form of mental retardation that caused, in the vast majority cases, by transcriptional silencing single gene, fmr1 . The encoded protein, FMRP, regulates mRNA translation neuronal dendrites, and it thought changes translation-dependent forms synaptic plasticity lead to many symptoms FXS. However, little known about potentially extensive protein content accompany loss FMRP. Here, we describe development high-throughput quantitative proteomic method identify differences expression between wild-type −/− mouse cortical neurons. based on stable isotope labeling amino acids cell culture (SILAC), which has been used characterize differentially expressed proteins dividing cells, but not terminally differentiated cells because reduced efficiency. To address issue incomplete labeling, developed mathematical normalize ratios relative reference Using this approach, conjunction with multidimensional identification technology (MudPIT), identified >100 are up- or down-regulated. These fall into variety functional categories, including those regulating structure, neurotransmission, dendritic transport, several implicated epilepsy autism, two endophenotypes studies provide insights potential origins abnormalities FXS demonstration methodology can be explore neurological disorders.