Investigation of mechanisms involved in perinatal asphyxia-induced cerebrovascular dysfunction is important because fetal and neonatal asphyxia can cause of perinatal brain damage and failure of blood flow regulation is a critical component. Carbon monoxide (CO) from astrocyte heme oxygenase (HO) is important in regulation of newborn cerebral circulation and preliminary data suggest HO products can provide cerebrovascular and astrocytic protection from asphyxia-induced damage. The research proposed pursues the unifying hypothesis that, in newborn brain, asphyxia disrupts astrocyte CO signaling and cerebral arteriolar smooth muscle (VSM) Ca2+ signaling and/or BKCa channel activity causing cerebrovascular dysfunction, while elevation of cytoprotective CO and bilirubin limit the dysfunction. To test this hypothesis, four specific hypotheses will be tested using a novel asphyxia model in newborn pigs: 1. Asphyxia injures astrocytes causing loss of CO-mediated dilation of pial arterioles, 2. Asphyxia depresses cerebral VSM CO-induced Ca2+ sparks, BKCa channel activity, and dilation, 3. Reactive oxygen species (ROS) are major contributors to asphyxia-induced astrocyte and VSM injury, and 4. Cytoprotective CO and bilirubin reduce asphyxia-induced ROS and post-asphyxia astrocyte and VSM dysfunction. Techniques will be used that allow investigation of intact newborn cerebral microcirculation in vivo, isolated pressurized cerebral arterioles, and freshly isolated astrocytes, microvessels and smooth muscle cells. Such research is unique by studying intact cerebral circulation, production of CO by intact brain and isolated astrocytes and microvessels, and investigating, at the cellular level, mechanisms responsible cerebral vascular dysfunction. In addition, experimental interventions that may attenuate cerebrovascular malfunction caused by asphyxia are examined. Cranial windows allow observation of microcirculation, collection of cortical CSF and topical application of agonists, precursors and inhibitors. CO production in vivo and in vitro is identified and quantified by gas chromatography-mass spectrometry. Astrocytes, cerebral arterioles, and cerebral arteriole smooth muscle cells isolated from control and post-asphyxia brains allow study of ROS, CO production, arteriolar reactivity, and vascular smooth muscle function. Global cytosolic Ca2+ and Ca2+ sparks in brain slices, intact arterioles and VSM will be studied using fluorescent indicator technology with a dual excitation, single emission system and laser scanning confocal microscopy, respectively. Patch clamp techniques will be used to examine BKCa channel activity. These studies are important because disorders of the cerebral circulation in the newborn period are major causes of sickness and death and can result in lifelong disabilities in survivors.

Public Health Relevance

Asphyxia may cause perinatal brain injury that leads to lifelong disabilities in survivors. The specific focus of this project is on the endogenous mechanisms that prevent loss of cerebral vascular function following asphyxia. Cerebral vascular dysfunction is often an important component in the development of neonatal brain injury. Understanding mechanisms of asphyxia-related loss of cerebral blood flow regulation is essential in improving the long-term neurological outcome of human newborns.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
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Pregnancy and Neonatology Study Section (PN)
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Reid, Diane M
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University of Tennessee Health Science Center
Schools of Medicine
United States
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Pourcyrous, Massroor; Chilakala, Sandeep; Elabiad, Mohamad T et al. (2018) Does prolonged severe hypercapnia interfere with normal cerebrovascular function in piglets? Pediatr Res 84:290-295
Liu, Jianxiong; Pourcyrous, Massroor; Fedinec, Alex L et al. (2017) Preventing harmful effects of epileptic seizures on cerebrovascular functions in newborn pigs: does sex matter? Pediatr Res 82:881-887
Harsono, Mimily; Pourcyrous, Massroor; Jolly, Elliott J et al. (2016) Selective head cooling during neonatal seizures prevents postictal cerebral vascular dysfunction without reducing epileptiform activity. Am J Physiol Heart Circ Physiol 311:H1202-H1213
Chang, Jennifer; Fedinec, Alexander L; Kuntamallappanavar, Guruprasad et al. (2016) Endothelial Nitric Oxide Mediates Caffeine Antagonism of Alcohol-Induced Cerebral Artery Constriction. J Pharmacol Exp Ther 356:106-15
Liu, Jianxiong; Fedinec, Alexander L; Leffler, Charles W et al. (2015) Enteral supplements of a carbon monoxide donor CORM-A1 protect against cerebrovascular dysfunction caused by neonatal seizures. J Cereb Blood Flow Metab 35:193-9
Pourcyrous, Massroor; Basuroy, Shyamali; Tcheranova, Dilyara et al. (2015) Brain-derived circulating endothelial cells in peripheral blood of newborn infants with seizures: a potential biomarker for cerebrovascular injury. Physiol Rep 3:
Nnorom, Chukwuma C; Davis, Corinne; Fedinec, Alexander L et al. (2014) Contributions of KATP and KCa channels to cerebral arteriolar dilation to hypercapnia in neonatal brain. Physiol Rep 2:
Bukiya, Anna; Dopico, Alejandro M; Leffler, Charles W et al. (2014) Dietary cholesterol protects against alcohol-induced cerebral artery constriction. Alcohol Clin Exp Res 38:1216-26
Basuroy, Shyamali; Leffler, Charles W; Parfenova, Helena (2013) CORM-A1 prevents blood-brain barrier dysfunction caused by ionotropic glutamate receptor-mediated endothelial oxidative stress and apoptosis. Am J Physiol Cell Physiol 304:C1105-15
Bukiya, Anna N; McMillan, Jacob E; Fedinec, Alexander L et al. (2013) Cerebrovascular dilation via selective targeting of the cholane steroid-recognition site in the BK channel ?1-subunit by a novel nonsteroidal agent. Mol Pharmacol 83:1030-44

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