Lipopolysaccharide stimulates BK channel activity in bladder umbrella cells

American Journal of Physiology. Cell Physiology
Ming LuToby C Chai

Abstract

Bladder urothelium plays an active role in response to bacterial infection. There is little known about the electrophysiological activity in urothelial cells in this process. We used a nonenzymatic method to isolate bladder urothelial tissue and to patch clamp umbrella cells in situ. A 200 pS conductance potassium (K+) channel was detected from female C57BL6 mice. Of 58 total patches, 17.2% patches displayed the 200 pS K+ conductance channel. This K+ conductance channel showed Ca2+ sensitivity and voltage dependence. Specific big-conductance potassium channel (BK) inhibitors (paxilline, iberiotoxin) blocked the 200 pS K+ conductance channel activity. RT-PCR and immunoblot confirmed BK channel pore-forming α-subunit (BK-α) mRNA and protein in urothelium. Immunohistochemistry also showed the BK-α located in urothelium. The above data provided evidence that the 200 pS K+ conductance channel was a BK channel. Lipopolysaccharide (LPS), a component of uropathogenic Escherichia coli, was used to investigate the role of BK channel in the pathogenesis of urinary tract infection. BK channel activity as NPo increased threefold within 30 min of exposure to LPS. mRNAs for LPS receptors (TLR4, CD14, MD-2) were expressed in the urothelium but...Continue Reading

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Citations

Mar 22, 2019·Nature Protocols·Ming LuToby C Chai
Oct 23, 2018·Biology of Sex Differences·Benjamin AbelsonMargot S Damaser
Mar 20, 2020·Physiological Reviews·Marianela G DalghiGerard Apodaca
Nov 21, 2020·American Journal of Respiratory Cell and Molecular Biology·Tatiana ZyrianovaAndreas Schwingshackl

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Methods Mentioned

BETA
reverse transcription PCR
PCR
dissection
electrophoresis

Software Mentioned

Clampfit
pCLAMP10
Image Lab
GraphPad Prism

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