Poly(ADP-ribose) polymerase (PARP) is an abundant nuclear enzyme which helps to maintain genomic integrity in neurons and numerous other cell types. Inhibition of the enzyme with pharmacological agents or PARP deficiency generated via transgenic paradigms result in reduced injury after cerebral ischemia. Our preliminary studies show that infarction after middle cerebral artery occlusion (MCAO) in PARP null mutants (PARP-/-) is strikingly reduced relative to wild type mice.
Aim 1 will fully examine this apparent neuroprotection observed in PARP deficient mice, characterizing extent and longevity of protection in the intact animal over the days of evolving infarction. We will use magnetic resonance imaging (MRI) to non-invasively examine the injured mouse brain and combine these data with functional behavior evaluation over time and terminal histopathology.
Aim 2 will establish the importance of nitric oxide generation to PARP activation in vivo and subsequent injury during MCAO. The relative consequences of inhibiting PARP versus neuronal or inducible nitric oxide synthase are examined in the mouse MCAO model. Lastly, using a novel strategy or transfecting virus carrying PARP mutant at two distinct catalytic and cleavage sites, we will begin to examine the molecular mechanism of PARP's important in stroke. The role of the C-terminal NAD binding domain in PARP- mediated neuroprotection will be determined by comparison of stroke outcomes in PARP null mice with or without transfection of a mutant form of PARP with inactive catalytic domain. In addition, we will examine cysteine apartase (CASPASE)-PARP interactions of comparison of stroke outcomes in PARP null mice with and without transfection of a mutant form of PARP lacking in active CASPASE 3 cleavage site (D214G) mutant. These experiments will further our understanding of PARP-mediated mechanisms of in vivo ischemic brain injury.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
5P01NS039148-03
Application #
6492587
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
2001-05-01
Project End
2002-04-30
Budget Start
Budget End
Support Year
3
Fiscal Year
2001
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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Wang, Yingfei; Kim, No Soo; Haince, Jean-Francois et al. (2011) Poly(ADP-ribose) (PAR) binding to apoptosis-inducing factor is critical for PAR polymerase-1-dependent cell death (parthanatos). Sci Signal 4:ra20
Andrabi, Shaida A; Kang, Ho Chul; Haince, Jean-François et al. (2011) Iduna protects the brain from glutamate excitotoxicity and stroke by interfering with poly(ADP-ribose) polymer-induced cell death. Nat Med 17:692-9
Kang, Ho Chul; Lee, Yun-Il; Shin, Joo-Ho et al. (2011) Iduna is a poly(ADP-ribose) (PAR)-dependent E3 ubiquitin ligase that regulates DNA damage. Proc Natl Acad Sci U S A 108:14103-8
Li, Xiaoling; Klaus, Judith A; Zhang, Jian et al. (2010) Contributions of poly(ADP-ribose) polymerase-1 and -2 to nuclear translocation of apoptosis-inducing factor and injury from focal cerebral ischemia. J Neurochem 113:1012-22
Xu, Zhenfeng; Zhang, Jian; David, Karen K et al. (2010) Endonuclease G does not play an obligatory role in poly(ADP-ribose) polymerase-dependent cell death after transient focal cerebral ischemia. Am J Physiol Regul Integr Comp Physiol 299:R215-21
Kishimoto, Koji; Li, Rung-Chi; Zhang, Jian et al. (2010) Cytosolic phospholipase A2 alpha amplifies early cyclooxygenase-2 expression, oxidative stress and MAP kinase phosphorylation after cerebral ischemia in mice. J Neuroinflammation 7:42
Wang, Yingfei; Kim, No S; Li, Xiaoling et al. (2009) Calpain activation is not required for AIF translocation in PARP-1-dependent cell death (parthanatos). J Neurochem 110:687-96
Yu, Seong-Woon; Wang, Yingfei; Frydenlund, Didrik S et al. (2009) Outer mitochondrial membrane localization of apoptosis-inducing factor: mechanistic implications for release. ASN Neuro 1:
Wang, Yingfei; Dawson, Valina L; Dawson, Ted M (2009) Poly(ADP-ribose) signals to mitochondrial AIF: a key event in parthanatos. Exp Neurol 218:193-202

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