Abstract
Genetic mutations of the Cl- channel ClC-5 cause Dent's disease in humans. We recently cloned an amphibian ortholog of Xenopus ClC-5 (xClC-5) from the A6 cell line. We now compare the properties and regulation of ClC-5 currents expressed in mammalian (COS-7) cells and Xenopus oocytes. Whole cell currents in COS-7 cells transfected with xClC-5 cDNA had strong outward rectification, Cl- > I- anion sensitivity, and were inhibited at low pH, similar to previous results in oocytes. In oocytes, antisense xClC-5 cRNA injection had no effect on endogenous membrane currents or the heterologous expression of human ClC-5. Activators of cAMP and protein kinase C inhibitors had no significant effects on ClC-5 currents expressed in either COS-7 cells or oocytes, whereas H-89, a cAMP-dependent protein kinase (PKA) inhibitor, and hydrogen peroxide decreased the currents. We conclude that the basic properties of ClC-5 currents were independent of the host cell type used for expression. In addition, ClC-5 channels may be modulated by PKA and reactive oxygen species.
| Original language | English (US) |
|---|---|
| Journal | American Journal of Physiology - Cell Physiology |
| Volume | 280 |
| Issue number | 6 49-6 |
| State | Published - 2001 |
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Keywords
- Dent's disease
- Hydrogen peroxide
- Mammalian COS-7 cells
- Patch clamp
- Xenopus oocytes
ASJC Scopus subject areas
- Clinical Biochemistry
- Cell Biology
- Physiology
- Physiology (medical)
Cite this
Expression and regulation of ClC-5 chloride channels : Effects of antisense and oxidants. / Weng, T. X.; Mo, L.; Hellmich, Helen; Yu, A. S L; Wood, Thomas; Wills, N. K.
In: American Journal of Physiology - Cell Physiology, Vol. 280, No. 6 49-6, 2001.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Expression and regulation of ClC-5 chloride channels
T2 - Effects of antisense and oxidants
AU - Weng, T. X.
AU - Mo, L.
AU - Hellmich, Helen
AU - Yu, A. S L
AU - Wood, Thomas
AU - Wills, N. K.
PY - 2001
Y1 - 2001
N2 - Genetic mutations of the Cl- channel ClC-5 cause Dent's disease in humans. We recently cloned an amphibian ortholog of Xenopus ClC-5 (xClC-5) from the A6 cell line. We now compare the properties and regulation of ClC-5 currents expressed in mammalian (COS-7) cells and Xenopus oocytes. Whole cell currents in COS-7 cells transfected with xClC-5 cDNA had strong outward rectification, Cl- > I- anion sensitivity, and were inhibited at low pH, similar to previous results in oocytes. In oocytes, antisense xClC-5 cRNA injection had no effect on endogenous membrane currents or the heterologous expression of human ClC-5. Activators of cAMP and protein kinase C inhibitors had no significant effects on ClC-5 currents expressed in either COS-7 cells or oocytes, whereas H-89, a cAMP-dependent protein kinase (PKA) inhibitor, and hydrogen peroxide decreased the currents. We conclude that the basic properties of ClC-5 currents were independent of the host cell type used for expression. In addition, ClC-5 channels may be modulated by PKA and reactive oxygen species.
AB - Genetic mutations of the Cl- channel ClC-5 cause Dent's disease in humans. We recently cloned an amphibian ortholog of Xenopus ClC-5 (xClC-5) from the A6 cell line. We now compare the properties and regulation of ClC-5 currents expressed in mammalian (COS-7) cells and Xenopus oocytes. Whole cell currents in COS-7 cells transfected with xClC-5 cDNA had strong outward rectification, Cl- > I- anion sensitivity, and were inhibited at low pH, similar to previous results in oocytes. In oocytes, antisense xClC-5 cRNA injection had no effect on endogenous membrane currents or the heterologous expression of human ClC-5. Activators of cAMP and protein kinase C inhibitors had no significant effects on ClC-5 currents expressed in either COS-7 cells or oocytes, whereas H-89, a cAMP-dependent protein kinase (PKA) inhibitor, and hydrogen peroxide decreased the currents. We conclude that the basic properties of ClC-5 currents were independent of the host cell type used for expression. In addition, ClC-5 channels may be modulated by PKA and reactive oxygen species.
KW - Dent's disease
KW - Hydrogen peroxide
KW - Mammalian COS-7 cells
KW - Patch clamp
KW - Xenopus oocytes
UR - http://www.scopus.com/inward/record.url?scp=0034992183&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034992183&partnerID=8YFLogxK
M3 - Article
VL - 280
JO - American Journal of Physiology - Endocrinology and Metabolism
JF - American Journal of Physiology - Endocrinology and Metabolism
SN - 0193-1849
IS - 6 49-6
ER -