Abstract

This proposal is a competing renewal application of our current project to study a role of Na+-K+-Cl- cotransporter isoform1 (NKCC1) in cerebral ischemic damage. The long-term goal of the research is to understand ischemia-induced cell death and develop a more effective approach to ischemia treatment. NKCC1 is important in regulation of intracellular Na+ and Cl-, cell volume, and K+ uptake in the central nervous system (CNS). In the initial funding period, we have investigated NKCC1 activity under several conditions that are associated with ischemic insults. We found the NKCC1 activity in cultured cortical astrocytes was significantly stimulated under high extracellular K+ ([K+]0). Pharmacological inhibition or genetic ablation of NKCC1 abolished high [K+]0-induced astrocyte swelling and decreased glutamate release. In cultured neurons, activation of both ionotropic and mGluR group l glutamate receptors stimulated NKCC1 activity in a Ca++-dependent manner. Infarct volume and cerebral edema were significantly reduced by bumetanide, a potent inhibitor for NKCC1. Lour preliminary study revealed a neuroprotection in NKCC1 knockout mice following focal ischemia. These data strongly suggest that NKCC1 has an important role in cerebral ischemic damage. However, the cellular mechanisms underlying the role of NKCC1 in ischemic cell damage have not been fully understood. We hypothesize that NKCC1 contributes to perturbation in ion homeostasis and necrotic ischemic cell death. We will test the hypothesis by following Specific Aims:
Aim 1 : Determine the role of NKCC1 in intracellular Na+ and Cl- overload, swelling, and the swelling-mediated glutamate release from cortical astrocytes in an in vitro oxygen and glucose deprivation (OGD) model of ischemia.
Aim 2 : Investigate the contribution of NKCC1 to OGD-mediated ischemic cell death in cortical neurons.
Aim 3 : Investigate whether genetic ablation of NKCC1 reduced brain damage (gray and white matter) in NKCC1-/- mice following transient focal ischemia.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS038118-04A1
Application #
6682184
Study Section
Special Emphasis Panel (ZRG1-BDCN-3 (01))
Program Officer
Michel, Mary E
Project Start
2000-02-01
Project End
2007-04-30
Budget Start
2003-07-15
Budget End
2004-04-30
Support Year
4
Fiscal Year
2003
Total Cost
$266,364
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Neurosurgery
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Zhao, Hanshu; Nepomuceno, Rachel; Gao, Xin et al. (2017) Deletion of the WNK3-SPAK kinase complex in mice improves radiographic and clinical outcomes in malignant cerebral edema after ischemic stroke. J Cereb Blood Flow Metab 37:550-563
Bhuiyan, Mohammad Iqbal H; Song, Shanshan; Yuan, Hui et al. (2017) WNK-Cab39-NKCC1 signaling increases the susceptibility to ischemic brain damage in hypertensive rats. J Cereb Blood Flow Metab 37:2780-2794
Yin, Yan; Sun, George; Li, Eric et al. (2017) ER stress and impaired autophagy flux in neuronal degeneration and brain injury. Ageing Res Rev 34:3-14
Boscia, Francesca; Begum, Gulnaz; Pignataro, Giuseppe et al. (2016) Glial Na(+) -dependent ion transporters in pathophysiological conditions. Glia 64:1677-97
Xu, Q; Deng, F; Xing, Z et al. (2016) Long non-coding RNA C2dat1 regulates CaMKII? expression to promote neuronal survival through the NF-?B signaling pathway following cerebral ischemia. Cell Death Dis 7:e2173
Roy, Ankita; Goodman, Joshua H; Begum, Gulnaz et al. (2015) Generation of WNK1 knockout cell lines by CRISPR/Cas-mediated genome editing. Am J Physiol Renal Physiol 308:F366-76
Cong, Damin; Zhu, Wen; Kuo, John S et al. (2015) Ion transporters in brain tumors. Curr Med Chem 22:1171-81
Zonouzi, Marzieh; Scafidi, Joseph; Li, Peijun et al. (2015) GABAergic regulation of cerebellar NG2 cell development is altered in perinatal white matter injury. Nat Neurosci 18:674-82
Adragna, Norma C; Ravilla, Nagendra B; Lauf, Peter K et al. (2015) Regulated phosphorylation of the K-Cl cotransporter KCC3 is a molecular switch of intracellular potassium content and cell volume homeostasis. Front Cell Neurosci 9:255
Begum, Gulnaz; Yuan, Hui; Kahle, Kristopher T et al. (2015) Inhibition of WNK3 Kinase Signaling Reduces Brain Damage and Accelerates Neurological Recovery After Stroke. Stroke 46:1956-1965

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