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PMID: 20826460 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

Regulation of bile canalicular network formation and maintenance by AMP-activated protein kinase and LKB1.

Journal of cell science ·Vol. 123 ·No. Pt 19 ·2010-10-01 ·Pages 3294-302

Fu D, Wakabayashi Y, Ido Y, Lippincott-Schwartz J, Arias IM

Abstract

AMP-activated protein kinase (AMPK), a cellular metabolic sensor, is essential in energy regulation and metabolism. Hepatocyte polarization during liver development and regeneration parallels increased metabolism. The current study investigates the effects of AMPK and its upstream activator LKB1 on polarity and bile canalicular network formation and maintenance in collagen sandwich cultures of rat hepatocytes. Immunostaining for the apical protein ABCB1 and the tight junction marker occludin demonstrated that canalicular network formation is sequential and is associated with activation of AMPK and LKB1. AMPK and LKB1 activators accelerated canalicular network formation. Inhibition of AMPK or LKB1 by dominant-negative AMPK or kinase-dead LKB1 constructs blocked canalicular network formation. AICAR and 2-deoxyglucose, which activate AMPK, circumvented the inhibitory effect of kinase-dead LKB1 on canalicular formation, indicating that AMPK directly affects canalicular network formation. After the canalicular network was formed, inhibition of AMPK and LKB1 by dominant-negative AMPK or kinase-dead LKB1 constructs resulted in loss of canalicular network, indicating that AMPK and LKB1 also participate in network maintenance. In addition, activation of AMPK and LKB1 prevented low-Ca(2+)-mediated disruption of the canalicular network and tight junctions. These studies reveal that AMPK and its upstream kinase, LKB1, regulate canalicular network formation and maintenance.

MeSH Terms
AMP-Activated Protein Kinase Kinases Animals Bile Canaliculi/growth & development,metabolism Cell Polarity/genetics Cells, Cultured Cloning, Molecular Enzyme Activation/genetics Hepatocytes/metabolism,pathology Male Mutant Proteins/genetics,metabolism Organ Culture Techniques Organogenesis/genetics Protein Kinases/genetics,metabolism Protein Serine-Threonine Kinases/genetics,metabolism Rats Rats, Sprague-Dawley
Chemicals
Mutant Proteins Protein Kinases Protein Serine-Threonine Kinases Stk11 protein, rat AMP-Activated Protein Kinase Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fu Dong
Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institute of Health, Bethesda, MD 20892, USA.
Wakabayashi Yoshiyuki
Ido Yasuo
Lippincott-Schwartz Jennifer
Arias Irwin M
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Article Info
Journal
Journal of cell science
Abbr.
J Cell Sci
ISSN
1477-9137
DOI
10.1242/jcs.068098
Published
2010-10-01
Epub
2010-00-07
Pages
3294-302
Language
English
Region
England
NLM ID
0052457
PMCID
PMC2939801
Subset
IM
Grants
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1242/jcs.068098 PMC Full Text View journal details
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