In Project 2, primary cortical neuronal, astroglial and mixed neuronal-astroglial cultures will be used to investigate whether proteins that show increased expression in an in vivo model of ischemic preconditioning and tolerance (Project 1) are cytoprotective in vitro. The hypothesis is that proteins showing increased expression in animal models of ischemic tolerance in vivo (Project 1) can be screened using cell culture models of ischemia and tolerance in vitro (Project 2) to identify proteins that will, in turn, protect neurons, astroglia or both from ischemia in vivo (Project 3).
The specific aims are to [1] determine which candidate cytoprotective proteins show increased expression in a cerebral cortical neuronal, astroglial or mixed neuronal-astroglial cell-culture model of tolerance to oxygen and glucose deprivation (OGD); [2] investigate which candidate cytoprotective proteins protect cultured cortical neurons, astroglia or both from otherwise lethal OGD, by determining the effect on cell viability of increasing their expression or (for secreted proteins) their extracellular concentration; [3] investigate which candidate cytoprotective proteins protect cultured cortical neurons, astroglia or both from otherwise lethal OGD, by determining the effect on cell viability of reducing their expression or inhibiting their function; and [4] investigate the mechanism of action of candidate cytoprotective proteins identified in Project I and confirmed in Specific Aims 1-3 of Project 2. The most promising candidate cytoprotective proteins that emerge from Project 2 will then be tested for protective effects in vivo in Project 3. The methods used will include primary culture of rat cerebral cortical neurons and astroglia; in vitro models of cerebral ischemia, preconditioning and tolerance; cell viability assays; Western blotting; immunocytochemistry; transfection with adeno-associated virus vectors; and transgenic and gene knockout mice. The broad, long-term objective is to identify endogenous protective mechanisms in ischemia and to exploit these in the treatment of stroke.
| Simon, Roger P (2016) Epigenetic modulation of gene expression governs the brain's response to injury. Neurosci Lett 625:16-9 |
| Simon, Roger P; Meller, Robert; Zhou, An et al. (2012) Can genes modify stroke outcome and by what mechanisms? Stroke 43:286-91 |
| Stevens, Susan L; Leung, Philberta Y; Vartanian, Keri B et al. (2011) Multiple preconditioning paradigms converge on interferon regulatory factor-dependent signaling to promote tolerance to ischemic brain injury. J Neurosci 31:8456-63 |
| Stapels, Martha; Piper, Chelsea; Yang, Tao et al. (2010) Polycomb group proteins as epigenetic mediators of neuroprotection in ischemic tolerance. Sci Signal 3:ra15 |
| Marsh, B J; Williams-Karnesky, R L; Stenzel-Poore, M P (2009) Toll-like receptor signaling in endogenous neuroprotection and stroke. Neuroscience 158:1007-20 |
| West, G Alexander; Golshani, Kiarash J; Doyle, Kristian P et al. (2009) A new model of cortical stroke in the rhesus macaque. J Cereb Blood Flow Metab 29:1175-86 |
| Stevens, Susan L; Ciesielski, Thomas M P; Marsh, Brenda J et al. (2008) Toll-like receptor 9: a new target of ischemic preconditioning in the brain. J Cereb Blood Flow Metab 28:1040-7 |
| Doyle, Kristian P; Yang, Tao; Lessov, Nikola S et al. (2008) Nasal administration of osteopontin peptide mimetics confers neuroprotection in stroke. J Cereb Blood Flow Metab 28:1235-48 |
| Khan, Adil A; Mao, Xiao Ou; Banwait, Surita et al. (2008) Regulation of hypoxic neuronal death signaling by neuroglobin. FASEB J 22:1737-47 |
| Pignataro, Giuseppe; Maysami, Samaneh; Studer, Francesca E et al. (2008) Downregulation of hippocampal adenosine kinase after focal ischemia as potential endogenous neuroprotective mechanism. J Cereb Blood Flow Metab 28:17-23 |
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