Visualization of G3BP Stress Granules Dynamics in Live Primary Cells
| Autor: | Martin, S., Chebli, K., Tazi, J. |
|---|---|
| Přispěvatelé: | Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Arsenites
Transgene Recombinant Fusion Proteins General Chemical Engineering Green Fluorescent Proteins Biology Cytoplasmic Granules Transfection SH3 domain General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Mice 0302 clinical medicine Stress granule stomatognathic system Stress Physiological Animals [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology RNA Messenger Poly-ADP-Ribose Binding Proteins 030304 developmental biology Genetics Neurons 0303 health sciences General Immunology and Microbiology Binding protein General Neuroscience Granule (cell biology) DNA Helicases Fibroblasts Embryonic stem cell Cell biology Cellular Biology RNA Recognition Motif Proteins Ribonucleoproteins Carrier Proteins 030217 neurology & neurosurgery Immunostaining RNA Helicases |
| Zdroj: | Journal of visualized experiments : JoVE Journal of visualized experiments : JoVE, JoVE, 2013, in press. ⟨10.3791/51197⟩ Journal of visualized experiments : JoVE, 2013, in press. ⟨10.3791/51197⟩ |
| ISSN: | 1940-087X |
| DOI: | 10.3791/51197⟩ |
| Popis: | SGs can be visualized in cells by immunostaining of specific protein components or polyA+ mRNAs. SGs are highly dynamic and the study of their assembly and fate is important to understand the cellular response to stress. The deficiency in key factors of SGs like G3BP (RasGAP SH3 domain Binding Protein) leads to developmental defects in mice and alterations of the Central Nervous System. To study the dynamics of SGs in cells from an organism, one can culture primary cells and follow the localization of a transfected tagged component of SGs. We describe time-lapse experiment to observe G3BP1-containing SGs in Mouse Embryonic Fibroblasts (MEFs). This technique can also be used to study G3BP-containing SGs in live neurons, which is crucial as it was recently shown that these SGs are formed at the onset of neurodegenerative diseases like Alzheimer's disease. This approach can be adapted to any other cellular body and granule protein component, and performed with transgenic animals, allowing the live study of granules dynamics for example in the absence of a specific factor of these granules. |
| Databáze: | OpenAIRE |
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