This is the second revision of a competitive renewal application to study basic mechanisms of hypoxic brain damage in newborn pigs. Experiments were designed to investigate specific cellular mechanisms of brain cell injury in the piglet by correlating the degree of hypoxia with alterations in the activity and structure of the N-methyl-D-aspartate (NMDA) receptor-ion channel complex and tissue histopathologic changes. The investigators hypothesize that the severity of hypoxia will have a specific impact on the function and possibly the structure of the NMDA receptor, which is a unique aspect of the grant application. The determination of the degree of hypoxia in vivo will be achieved by the continuous monitoring of brain high energy phosphate reserves using 31-P nuclear magnetic resonance spectroscopy as well as the determination of cerebral cortical oxygen pressure by oxygen- dependent quenching of phosphorescence. Measurements of cell membrane lipid peroxidation, sodium/potassium- ATPase activity and the presence of oxygen free radicals will be used as indices of brain cell injury. Light microscopy will be performed to assess histopathologic changes at 48 hours of recovery from hypoxia. All experiments will be conducted in newborn pigs to investigate: 1) the relationship between quantitative tissue hypoxia and the structure of the recognition and modulatory sites and the activation of the receptor/ion channel complex; 2) the effect of hypoxia on NMDA-mediated influx of calcium into synaptoneurosomes; 3) the relationship between the occurrence of lipid peroxidation and alterations in the recognition and modulatory sites of the NMDA receptor; 4) the relationship between quantitative tissue hypoxia and the release of specific monoamines and amino acid neurotransmitters; and 5) the effect of increased release of amino acid and monoamine neurotransmitters during hypoxia on the activity of brain cell sodium/potassium-ATPase and alterations in NMDA activity.Additional experiments will investigate the potentiating effect of hypoglycemia during hypoxia on both NMDA receptor activity and cell membrane function. The proposed experiments will be performed using well- established techniques for the determination of ligand binding and enzyme kinetics, microdialysis, HPLC, histopathologic methods and others. It is proposed that the findings of the experiments will provide new insights into brain cell membrane function and will lead to a better understanding of basic mechanisms contributing to hypoxic neuronal injury in the newborn human infant.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD020337-13
Application #
2643521
Study Section
Human Embryology and Development Subcommittee 1 (HED)
Project Start
1985-09-01
Project End
1998-11-30
Budget Start
1997-12-01
Budget End
1998-11-30
Support Year
13
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Allegheny University of Health Sciences
Department
Pediatrics
Type
Schools of Medicine
DUNS #
City
Philadelphia
State
PA
Country
United States
Zip Code
19129
Kratimenos, Panagiotis; Koutroulis, Ioannis; Jain, Amit et al. (2018) Effect of Concurrent Src Kinase Inhibition with Short-Duration Hypothermia on Ca2+/Calmodulin Kinase IV Activity and Neuropathology after Hypoxia-Ischemia in the Newborn Swine Brain. Neonatology 113:37-43
Delivoria-Papadopoulos, Maria (2012) Mechanism of caspase-9 activation during hypoxia in the cerebral cortex of newborn piglets: the role of Src kinase. Neurosci Lett 523:19-23
Delivoria-Papadopoulos, Maria; Ashraf, Qazi M; Mishra, Om Prakash (2011) Mechanism of CaM kinase IV activation during hypoxia in neuronal nuclei of the cerebral cortex of newborn piglets: the role of Src kinase. Neurochem Res 36:1512-9
Delivoria-Papadopoulos, Maria; Ashraf, Qazi M; Mishra, Om Prakash (2011) Brain tissue energy dependence of CaM kinase IV cascade activation during hypoxia in the cerebral cortex of newborn piglets. Neurosci Lett 491:113-7
Delivoria-Papadopoulos, Maria; Mishra, Om Prakash (2010) Mechanism of post-translational modification by tyrosine phosphorylation of apoptotic proteins during hypoxia in the cerebral cortex of newborn piglets. Neurochem Res 35:76-84
Mishra, Om Prakash; Ashraf, Qazi M; Delivoria-Papadopoulos, Maria (2010) Hypoxia-induced activation of epidermal growth factor receptor (EGFR) kinase in the cerebral cortex of newborn piglets: the role of nitric oxide. Neurochem Res 35:1471-7
Mishra, Om P; Delivoria-Papadopoulos, Maria (2010) Mechanism of tyrosine phosphorylation of procaspase-9 and Apaf-1 in cytosolic fractions of the cerebral cortex of newborn piglets during hypoxia. Neurosci Lett 480:35-9
Mudduluru, Manjula; Zubrow, Alan B; Ashraf, Q M et al. (2010) Tyrosine phosphorylation of apoptotic proteins during hyperoxia in mitochondria of the cerebral cortex of newborn piglets. Neurochem Res 35:1003-9
Mishra, Om Prakash; Ashraf, Qazi M; Delivoria-Papadopoulos, Maria (2010) Mechanism of increased tyrosine (Tyr(99)) phosphorylation of calmodulin during hypoxia in the cerebral cortex of newborn piglets: the role of nNOS-derived nitric oxide. Neurochem Res 35:67-75
Mishra, Om Prakash; Ashraf, Qazi M; Delivoria-Papadopoulos, Maria (2009) NO-mediated activation of Src kinase during hypoxia in the cerebral cortex of newborn piglets. Neurosci Lett 460:61-5

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