Proteomic assessment of a cell model of spinal muscular atrophy
| Autor: | Douglas A. Kerr, Dosh Whye, Robert W. Mason, Lisa Glazewski, Wenlan Wang, Kelvin H. Lee, Chia-Yen Wu, Leila Choe |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Proteomics
Pathology Aging Proteome Cellular differentiation SMN1 Mice 0302 clinical medicine Sonic hedgehog Cell Line Transformed Motor Neurons 0303 health sciences General Neuroscience Stem Cells lcsh:QP351-495 Cell Differentiation SMA Cell biology Neuromuscular diseases medicine.anatomical_structure Spinal Cord Stem cell Research Article Morphology medicine.medical_specialty Fibroblast Growth Factor 8 Green Fluorescent Proteins Mice Transgenic Nerve Tissue Proteins Protein degradation Biology Antibodies lcsh:RC321-571 Muscular Atrophy Spinal 03 medical and health sciences Cellular and Molecular Neuroscience medicine Animals lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry 030304 developmental biology Spinal muscular atrophy Motor neuron medicine.disease Embryo Mammalian Survival of Motor Neuron 1 Protein Fibroblast Growth Factors Disease Models Animal lcsh:Neurophysiology and neuropsychology Gene Expression Regulation nervous system biology.protein Proteins--Analysis 030217 neurology & neurosurgery |
| Zdroj: | BMC Neuroscience, Vol 12, Iss 1, p 25 (2011) BMC Neuroscience |
| DOI: | 10.7916/d8c53kkz |
| Popis: | Background Deletion or mutation(s) of the survival motor neuron 1 (SMN1) gene causes spinal muscular atrophy (SMA), a neuromuscular disease characterized by spinal motor neuron death and muscle paralysis. Complete loss of the SMN protein is embryonically lethal, yet reduced levels of this protein result in selective death of motor neurons. Why motor neurons are specifically targeted by SMN deficiency remains to be determined. In this study, embryonic stem (ES) cells derived from a severe SMA mouse model were differentiated into motor neurons in vitro by addition of retinoic acid and sonic hedgehog agonist. Proteomic and western blot analyses were used to probe protein expression alterations in this cell-culture model of SMA that could be relevant to the disease. Results When ES cells were primed with Noggin/fibroblast growth factors (bFGF and FGF-8) in a more robust neural differentiation medium for 2 days before differentiation induction, the efficiency of in vitro motor neuron differentiation was improved from ~25% to ~50%. The differentiated ES cells expressed a pan-neuronal marker (neurofilament) and motor neuron markers (Hb9, Islet-1, and ChAT). Even though SMN-deficient ES cells had marked reduced levels of SMN (~20% of that in control ES cells), the morphology and differentiation efficiency for these cells are comparable to those for control samples. However, proteomics in conjunction with western blot analyses revealed 6 down-regulated and 14 up-regulated proteins with most of them involved in energy metabolism, cell stress-response, protein degradation, and cytoskeleton stability. Some of these activated cellular pathways showed specificity for either undifferentiated or differentiated cells. Increased p21 protein expression indicated that SMA ES cells were responding to cellular stress. Up-regulation of p21 was confirmed in spinal cord tissues from the same SMA mouse model from which the ES cells were derived. Conclusion SMN-deficient ES cells provide a cell-culture model for SMA. SMN deficiency activates cellular stress pathways, causing a dysregulation of energy metabolism, protein degradation, and cytoskeleton stability. |
| Databáze: | OpenAIRE |
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