Publication:
P2Y receptor regulation of K2p channels that facilitate K + secretion by human mammary epithelial cells

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Issued Date

2018

Resource Type

Article

File Type

application/pdf

ISSN

3636143

DOI

10.1152/ajpcell.00342.2016

Other identifier(s)

2-s2.0-85061338059

Rights Holder(s)

Scopus

Bibliographic Citation

American Journal of Physiology - Cell Physiology. Vol 314, No.5 (2018), p.C627-C639

Suggested Citation

Srisomboon Y., Zaidman N.A., Maniak P.J., Deachapunya C., O’Grady S.M. P2Y receptor regulation of K2p channels that facilitate K + secretion by human mammary epithelial cells. American Journal of Physiology - Cell Physiology. Vol 314, No.5 (2018), p.C627-C639. doi:10.1152/ajpcell.00342.2016 Retrieved from: https://hdl.handle.net/20.500.14740/3954

Title

P2Y receptor regulation of K2p channels that facilitate K + secretion by human mammary epithelial cells

Author(s)

Srisomboon Y.
 Zaidman N.A.
 Maniak P.J.
 Deachapunya C.
 O’Grady S.M.

Abstract

The objective of this study was to determine the molecular identity of ion channels involved in K + secretion by the mammary epithelium and to examine their regulation by purinoceptor agonists. Apical membrane voltage-clamp experiments were performed on human mammary epithelial cells where the basolateral membrane was exposed to the pore-forming antibiotic amphotericin B dissolved in a solution with intracellular-like ionic composition. Addition of the Na + channel inhibitor benzamil reduced the basal current, consistent with inhibition of Na + uptake across the apical membrane, whereas the K Ca 3.1 channel blocker TRAM-34 produced an increase in current resulting from inhibition of basal K + efflux. Treatment with two-pore potassium (K2P) channel blockers quinidine, bupivacaine and a selective TASK1/TASK3 inhibitor (PK-THPP) all produced concentration-dependent inhibition of apical K + efflux. qRT-PCR experiments detected mRNA expression for nine K2P channel subtypes. Western blot analysis of biotinylated apical membranes and confocal immunocytochemistry revealed that at least five K2P subtypes (TWIK1, TREK1, TREK2, TASK1, and TASK3) are expressed in the apical membrane. Apical UTP also increased the current, but pretreatment with the PKC inhibitor GF109203X blocked the response. Similarly, direct activation of PKC with phorbol 12-myristate 13-acetate produced a similar increase in current as observed with UTP. These results support the conclusion that the basal level of K + secretion involves constitutive activity of apical K Ca 3.1 channels and multiple K2P channel subtypes. Apical UTP evoked a transient increase in K Ca 3.1 channel activity, but over time caused persistent inhibition of K2P channel function leading to an overall decrease in K + secretion. © 2018 the American Physiological Society.

Subject(s)

Epithelial sodium channel
 KCa3.1 channel
 Messenger RNA
 Potassium channel
 Protein kinase C
 Purinergic P2Y receptor
 TASK1 channel
 TASK3 channel
 TREK1 channel
 TREK2 channel
 TWIK1 channel
 Two pore potassium channel
 Unclassified drug
 Epithelial sodium channel
 Potassium
 Potassium channel blocking agent
 Protein kinase C
 Purinergic P2Y receptor
 Purinergic P2Y receptor agonist
 Sodium
 Tandem pore domain potassium channel
 Uridine triphosphate
 Apical membrane
 Article
 Breast epithelium cell
 Cellular distribution
 Controlled study
 Human
 Human cell
 Immunocytochemistry
 Lipid raft
 Membrane current
 Potassium transport
 Priority journal
 Reverse transcription polymerase chain reaction
 Sodium absorption
 Sodium transport
 Voltage clamp technique
 Antagonists and inhibitors
 Cytology
 Drug effect
 Epithelium cell
 Female
 Genetics
 Mammary gland
 Membrane potential
 Metabolism
 Secretory pathway
 Transformed cell line
 Cell Line, Transformed
 Epithelial Cells
 Epithelial Sodium Channels
 Female
 Humans
 Mammary Glands, Human
 Membrane Potentials
 Potassium
 Potassium Channel Blockers
 Potassium Channels, Tandem Pore Domain
 Protein Kinase C
 Purinergic P2Y Receptor Agonists
 Receptors, Purinergic P2Y
 Secretory Pathway
 Sodium
 Uridine Triphosphate

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URI

https://hdl.handle.net/20.500.14740/3954

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Scopus: Year 1983-2021

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