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Characterizing the role of the RNA-binding protein RBM3 in neuroprotection and neuronal cell differentiation

English title Characterizing the role of the RNA-binding protein RBM3 in neuroprotection and neuronal cell differentiation
Applicant Wellmann Sven
Number 141007
Funding scheme Project funding (Div. I-III)
Research institution Abteilung Neonatologie Universitäts-Kinderspital beider Basel
Institution of higher education University of Basel - BS
Main discipline Neurophysiology and Brain Research
Start/End 01.07.2012 - 31.08.2015
Approved amount 302'958.00
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All Disciplines (2)

Discipline
Neurophysiology and Brain Research
Molecular Biology

Keywords (3)

Hypothermia; Neuroprotection; Neurodifferentiation

Lay Summary (English)

Lead
Lay summary
Neonatal asphyxia with subsequent brain damage, known as hypoxic-ischemic encephalopathy, occurs in 0.5 - 2 % of all term births despite ongoing advances in obstetrical and neonatal care. Whole body hypothermia of 33°C for three days is the only therapy with proven efficacy to reduce neurodevelopmental impairment. Whereas hypothermia reduces global protein synthesis and cell metabolism, low temperature induces a small subset of proteins that includes the RNA-binding protein RBM3. This protein plays a role in hibernation, the seasonally entrained and profound phenotypic transition to conserve energy in winter used by a wide range of mammalian species. We have just recently demonstrated a direct link between RBM3 and hypothermia-induced neuroprotection in isolated cells and in organotypic brain slice cultures. We hypothesize a critical role for RBM3 in neuronal cell growth at least in recovery from damage and are investigating this by use of various in-vitro and in-vivo models including RBM3 transgenic mice.
Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

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Associated projects

Number Title Start Funding scheme
163305 Cold-inducible RBM3 in neuroprotection and neurogenesis 01.10.2015 Project funding (Div. I-III)
124101 From cortex to classroom: enhancing brain development for premature infants 01.10.2009 SPUM

Abstract

Background: Induced hypothermia is the only therapy with proven efficacy to reduce brain damage after perinatal asphyxia. While hypothermia reduces global protein synthesis and cell metabolism, low temperature induces a small subset of proteins that includes the RNA-binding protein RBM3. We have just recently demonstrated a direct link between RBM3 and hypothermia-induced neuroprotection in isolated cells and in organotypic brain slice cultures (OBSCs). Experiments carried out to elucidate the protective mechanisms in our laboratory point to a previously not described critical physical interaction between nuclear proteins with RBM3. Working hypotheses: (1) RBM3 is important in neuroprotection afforded by induced hypothermia.(2) RBM3 acts by recruiting specific proteins (protein-protein interaction) and by stabilization of distinct anti-apoptotic mRNA species.(3) RBM3 plays a critical role in neuronal cell growth at least in recovery from damage. Specific aims, objectives: (1)to verify interaction of RBM3 with putative interacting proteins.(2) to study brain damage, challenged by oxygen-glucose deprivation and excitotoxicity, together with hypothermia on cell integrity, RBM3 and putative RBM3 interactors in OBSCs.(3)to determine functions of RBM3 and interacting proteins in neuronal cell differentiation.(4) to compare postnatal neurodevelopment between RBM3-/- and wild-type mice.(5) to determine brain damage and hypothermia-induced neuroprotection in OBSCs obtained from RBM3 -/- mice. Experimental methods: (1) Inducible Tet-on-HEK293 and Tet-on-PC12 cells; OBSCs of control and RBM3-/- mice(2) siRNA mediated gene silencing, co-immunoprecipitation, immunoblotting, quantitative RT-PCR, flow cytometry to determine ratios of viable, apoptotic and necrotic cells as well as cell cycle; MTT, LDH assays, cDNA-cloning and gene expression.(3) Colocalization by fluorescence immunohistochemistry and confocal laser microscopy.(4) Nissl stain, BrdU-labeling, staining of various neuronal markers and cortical layers.Expected value of project: To enhance the understanding of the biological role of RBM3, with a special focus on enhanced neuronal cell survival mediated by induced hypothermia.
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