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Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes

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Contribution of KCNQ1 to the regulatory volume decrease in the human mammary epithelial cell line MCF-7.

Brenna L vanTol¹, Sergey Missan, Julie Crack

¹Dept. of Physiology and Biophysics, Dalhousie Univ., Halifax, Nova Scotia B3H 1X5, Canada. elizabeth.cowley@dal.ca).

American Journal of Physiology. Cell Physiology

|June 29, 2007

Summary

The KCNQ1 potassium channel is crucial for regulating cell volume in human mammary epithelial cells (MCF-7) during hyposmotic stress, facilitating regulatory volume decrease.

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Area of Science:

Cell Biology
Physiology
Ion Channel Function

Background:

Cell volume regulation is essential for cellular homeostasis.
Mammary epithelial cells have specific ion transport needs.
The role of KCNQ1 channels in mammary cell volume control was unclear.

Purpose of the Study:

To investigate the role of KCNQ1 channels in hyposmotic stress response in MCF-7 cells.
To determine the localization and function of KCNQ1 in mammary epithelial cell volume regulation.

Main Methods:

MCF-7 cells were subjected to hyposmotic stress.
Pharmacological inhibitors (Ba(2+), quinine, TEA, chromonol 293B, XE991) were used to block K(+) channels.
Dominant-negative KCNQ1 isoforms were employed.
KCNQ1 expression was confirmed via mRNA and protein analysis.
Immunofluorescence localized KCNQ1 in polarized monolayers.
Whole-cell patch-clamp electrophysiology was performed.

Main Results:

MCF-7 cells exhibited regulatory volume decrease (RVD) under hyposmotic conditions.
K(+) channel inhibitors significantly blocked the RVD response.
Specific KCNQ1 inhibitors and dominant-negative isoforms abolished RVD.
KCNQ1 mRNA and protein are expressed in MCF-7 cells.
KCNQ1 localizes to the apical membrane in polarized MCF-7 monolayers.
A hyposmotic-induced, 293B-sensitive K(+) current was detected in MCF-7 cells.
Heterologous expression confirmed KCNQ1/KCNE3 forms a volume-sensitive K(+) current.

Conclusions:

KCNQ1 channels are critical for regulating cell volume in MCF-7 cells during hyposmotic stress.
KCNQ1 plays a significant role in the RVD mechanism of mammary epithelial cells.
KCNQ1 may mediate transepithelial potassium secretion in the mammary epithelium.