Abstract

nA+-k+-2Cl-cotransporters (NKCC) are important in renal salt reabsorption and secretion by reabsorptive and secretory epithelia. The NKCC also function in maintenance and regulation of cell volume in both epithelial and non-epithelial cells. However, the role of the NKCC in the central nervous system (CNS) have not been defined. The long-term goal of this research is to understand function and regulation of the NKCC in the CNS under physiological as well as pathological conditions such as ischemia. Our pilot studies described in this proposal suggest that the NKCC are essential in maintenance of K+ homeostasis in astrocytes and they also appear to be involved in glutamate (Glu)-mediated neurotoxicity. There are four Specific Aims to test our hpotheses.
Aim 1 : Characterize high [K+]o-mediated stimulation of the NKCC activity in primary astrocyte cultures. The objectives are: 1). to verify that high [K+]o causes stimulation of the NKCC; 2). to determine whether the high [K+]o mediated stimulation of the NKCC is due to an increase in intracellular [Ca++].
Aim 2 : Determine whether high [K+]o- mediated stimulation of the NKCC causes astrocyte swelling and non-vesicular release of Glu. The objectives are to investigate: 1). whether the NKCC contributes to accumulation of intracellular Na+, C1 and cell swelling under [K+]o; 2). whether inhibition of the NKCC activity could significantly increase [3H]-L-Glu uptake, while blocking of high [K+]-evoked release of Glu from astrocytes.
Aim 3 : Investigate whether stimulation of the NKCC contributes to Glu-mediated neuronal toxicity. The objectives are to investigate: 1). whether the NKCC contributes to Glu- mediated increase in [C1-]i and neuronal swelling; 2). whether inhibition of the NKCC activity could protect cells from Glu- induced damage.
Aim 4 : Determine whether ischemia-induced cell damage is reduced when the NKCC activity is transiently inhibited by a local administration of bumetanide in brain. This study will be performed in an in vivo cerebral ischemic model.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS038118-02
Application #
6351873
Study Section
Special Emphasis Panel (ZRG1-BDCN-3 (01))
Program Officer
Behar, Toby
Project Start
2000-02-01
Project End
2003-01-31
Budget Start
2001-02-01
Budget End
2002-01-31
Support Year
2
Fiscal Year
2001
Total Cost
$191,618
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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