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Astrocytes from Na+-K+-Cl−cotransporter-null mice exhibit absence of swelling and decrease in EAA release

Su, Gui, Kintner, Douglas B., Flagella, Michael, Shull, Gary E., Sun, Dandan·2002·American Journal of Physiology-Cell Physiology
DOI 10.1152/ajpcell.00538.2001
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Abstract

We reported previously that inhibition of Na+-K+-Cl− cotransporter isoform 1 (NKCC1) by bumetanide abolishes high extracellular K+concentration ([K+]o)-induced swelling and intracellular Cl− accumulation in rat cortical astrocytes. In this report, we extended our study by using cortical astrocytes from NKCC1-deficient (NKCC1−/−) mice. NKCC1 protein and activity were absent in NKCC1−/− astrocytes. [K+]o of 75 mM increased NKCC1 activity approximately fourfold in NKCC1+/+ cells ( P< 0.05) but had no effect in NKCC1−/− astrocytes. Intracellular Cl− was increased by 70% in NKCC1+/+ astrocytes under 75 mM [K+]o ( P < 0.05) but remained unchanged in NKCC1−/− astrocytes. Baseline intracellular Na+ concentration ([Na+]i) in NKCC1+/+ astrocytes was 19.0 ± 0.5 mM, compared with 16.9 ± 0.3 mM [Na+]i in NKCC1−/− astrocytes ( P < 0.05). Relative cell volume of NKCC1+/+ astrocytes increased by 13 ± 2% in 75 mM [K+]o, compared with a value of 1.0 ± 0.5% in NKCC1−/− astrocytes ( P < 0.05). Regulatory volume increase after hypertonic shrinkage was completely impaired in NKCC1−/− astrocytes. High-[K+]o-induced 14 C-labeledd-aspartate release was reduced by ∼30% in NKCC1−/− astrocytes. Our study suggests that stimulation of NKCC1 is required for high-[K+]o-induced swelling, which contributes to glutamate release from astrocytes under high [K+]o.

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Crossref · 10.1152/ajpcell.00538.2001

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Authors
Su, G., Kintner, D. B., Flagella, M., Shull, G. E., & Sun, D.
Affiliations
  • University of Wisconsin–Madison (US)
  • Departments of Neurosurgery and
  • University of Cincinnati (US)
ORCID
Venue
American Journal of Physiology-Cell Physiology
Publisher
American Physiological Society
Published
2002-05-01
Volume
282
Issue
5
Pages
C1147-C1160
ISSN
  • 0363-6143 (ISSN-L)
  • 1522-1563
DOI
10.1152/ajpcell.00538.2001
OpenAlex
W2103230981
Cited by
169
References
42
Citations by year
2025: 2 · 2024: 7 · 2023: 5 · 2022: 7 · 2021: 8 · 2020: 14 · 2019: 9 · 2018: 5 · 2017: 8 · 2016: 5 · 2015: 11 · 2014: 11 · 2013: 5 · 2012: 10
FWCI
2.44
Type
journal article
Language
en
Open access
closed
Locations
  • American Journal of Physiology-Cell Physiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institute of Diabetes and Digestive and Kidney Diseases (R01DK050594, R01DK-50594)
  • National Institute of Neurological Disorders and Stroke (R01-NS-38118, R01NS038118)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Amino Acid Enzymes and Metabolism
Keywords
Na+-K+-2Cl- cotransporter, NKCC1, astrocytes, cell swelling, high extracellular potassium, intracellular sodium, glutamate release, D-aspartate, bumetanide, cortical astrocytes
MeSH terms
Animals, Aspartic Acid, Astrocytes, Bumetanide, Cells, Cultured, Cerebral Cortex, Chlorides, Diuretics, Microscopy, Fluorescence, Potassium, Sodium, 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid, Mice, Knockout, Glutamic Acid, Sodium-Potassium-Chloride Symporters, Cell Size, Mice, Rats

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