Endogenous TRPM4-like channel in Chinese hamster ovary (CHO) cells

Oleg V. Yarishkin; Eun Mi Hwang; Jae Yong Park; Dawon Kang; Jaehee Han; Seong Geun Hong

doi:10.1016/j.bbrc.2008.02.081

Endogenous TRPM4-like channel in Chinese hamster ovary (CHO) cells

Oleg V. Yarishkin, Eun Mi Hwang, Jae Yong Park, Dawon Kang, Jaehee Han, Seong Geun Hong

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

Abstract

Chinese hamster ovary (CHO) cells used in many transfection studies have been found to endogenously express channels permeable to monovalent cations, but not to divalent cations. In the presence of intracellular Ca²⁺, 23-pS channel with a linear current-voltage (I-V) relationship could be frequently observed in inside-out patches but not in cell-attached patches. The open probability was voltage-dependent, which is higher at positive potentials. The channel was dose-dependently activated by relatively high level of Ca²⁺ (EC₅₀ = 1.04 ± 0.08 mM), and sensitively inhibited by 100 μM ATP, ADP, AMP, and 1 mM spermine. However, ruthenium red (2 μM) had no effect. Reverse transcript polymerase chain reaction (RT-PCR) supported the presence of mRNA encoding TRPM4b channel protein. Western blot assay finally confirmed the presence of this channel protein in membrane fraction of CHO cells. These results provide evidence that CHO cells express an endogenous TRPM4b-like channel, and thereby can be used as a tool to study de novo regulation/modulation of TRPM4 channel.

Original language	English
Pages (from-to)	712-717
Number of pages	6
Journal	Biochemical and biophysical research communications
Volume	369
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2008.02.081
Publication status	Published - 2008 May 2
Externally published	Yes

Keywords

CHO cell
Nonselective cation channel
TRPM4b

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2008.02.081

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title = "Endogenous TRPM4-like channel in Chinese hamster ovary (CHO) cells",

abstract = "Chinese hamster ovary (CHO) cells used in many transfection studies have been found to endogenously express channels permeable to monovalent cations, but not to divalent cations. In the presence of intracellular Ca2+, 23-pS channel with a linear current-voltage (I-V) relationship could be frequently observed in inside-out patches but not in cell-attached patches. The open probability was voltage-dependent, which is higher at positive potentials. The channel was dose-dependently activated by relatively high level of Ca2+ (EC50 = 1.04 ± 0.08 mM), and sensitively inhibited by 100 μM ATP, ADP, AMP, and 1 mM spermine. However, ruthenium red (2 μM) had no effect. Reverse transcript polymerase chain reaction (RT-PCR) supported the presence of mRNA encoding TRPM4b channel protein. Western blot assay finally confirmed the presence of this channel protein in membrane fraction of CHO cells. These results provide evidence that CHO cells express an endogenous TRPM4b-like channel, and thereby can be used as a tool to study de novo regulation/modulation of TRPM4 channel.",

keywords = "CHO cell, Nonselective cation channel, TRPM4b",

author = "Yarishkin, {Oleg V.} and Hwang, {Eun Mi} and Park, {Jae Yong} and Dawon Kang and Jaehee Han and Hong, {Seong Geun}",

note = "Funding Information: This work was supported by Grant No. (R13-2005-012-01002-0) from the Basic Research Program of the KOSEF. Oleg V. Yarishkin was supported by Korea Research Foundation Grant (KRF-2004-005-E00008).",

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AU - Yarishkin, Oleg V.

AU - Hwang, Eun Mi

AU - Park, Jae Yong

AU - Kang, Dawon

AU - Han, Jaehee

AU - Hong, Seong Geun

N1 - Funding Information: This work was supported by Grant No. (R13-2005-012-01002-0) from the Basic Research Program of the KOSEF. Oleg V. Yarishkin was supported by Korea Research Foundation Grant (KRF-2004-005-E00008).

PY - 2008/5/2

Y1 - 2008/5/2

N2 - Chinese hamster ovary (CHO) cells used in many transfection studies have been found to endogenously express channels permeable to monovalent cations, but not to divalent cations. In the presence of intracellular Ca2+, 23-pS channel with a linear current-voltage (I-V) relationship could be frequently observed in inside-out patches but not in cell-attached patches. The open probability was voltage-dependent, which is higher at positive potentials. The channel was dose-dependently activated by relatively high level of Ca2+ (EC50 = 1.04 ± 0.08 mM), and sensitively inhibited by 100 μM ATP, ADP, AMP, and 1 mM spermine. However, ruthenium red (2 μM) had no effect. Reverse transcript polymerase chain reaction (RT-PCR) supported the presence of mRNA encoding TRPM4b channel protein. Western blot assay finally confirmed the presence of this channel protein in membrane fraction of CHO cells. These results provide evidence that CHO cells express an endogenous TRPM4b-like channel, and thereby can be used as a tool to study de novo regulation/modulation of TRPM4 channel.

AB - Chinese hamster ovary (CHO) cells used in many transfection studies have been found to endogenously express channels permeable to monovalent cations, but not to divalent cations. In the presence of intracellular Ca2+, 23-pS channel with a linear current-voltage (I-V) relationship could be frequently observed in inside-out patches but not in cell-attached patches. The open probability was voltage-dependent, which is higher at positive potentials. The channel was dose-dependently activated by relatively high level of Ca2+ (EC50 = 1.04 ± 0.08 mM), and sensitively inhibited by 100 μM ATP, ADP, AMP, and 1 mM spermine. However, ruthenium red (2 μM) had no effect. Reverse transcript polymerase chain reaction (RT-PCR) supported the presence of mRNA encoding TRPM4b channel protein. Western blot assay finally confirmed the presence of this channel protein in membrane fraction of CHO cells. These results provide evidence that CHO cells express an endogenous TRPM4b-like channel, and thereby can be used as a tool to study de novo regulation/modulation of TRPM4 channel.

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KW - Nonselective cation channel

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Endogenous TRPM4-like channel in Chinese hamster ovary (CHO) cells

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Keywords

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