Document Detail


Imaging and characterization of stretch-induced ATP release from alveolar A549 cells.
MedLine Citation:
PMID:  23247110     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Abstract  Mechano-transduction at cellular and tissue levels often involves ATP release and activation of the purinergic signalling cascade. In the lungs, stretch is an important physical stimulus but its impact on ATP release, the underlying release mechanisms and transduction pathways are poorly understood. Here, we investigated the effect of unidirectional stretch on ATP release from human alveolar A549 cells by real-time luciferin-luciferase bioluminescence imaging coupled with simultaneous infrared imaging, to monitor the extent of cell stretch and to identify ATP releasing cells. In subconfluent (<90%) cell cultures, single 1 s stretch (10-40%)-induced transient ATP release from a small fraction (1.5%) of cells that grew in number dose-dependently with increasing extent of stretch. ATP concentration in the proximity (150 μm) of releasing cells often exceeded 10 μm, sufficient for autocrine/paracrine purinoreceptor stimulation of neighbouring cells. ATP release responses were insensitive to the putative ATP channel blockers carbenoxolone and 5-nitro-2-(3-phenylpropyl-amino) benzoic acid, but were inhibited by N-ethylmaleimide and bafilomycin. In confluent cell cultures, the maximal fraction of responding cells dropped to <0.2%, but was enhanced several-fold in the wound/scratch area after it was repopulated by new cells during the healing process. Fluo8 fluorescence experiments revealed two types of stretch-induced intracellular Ca(2+) responses, rapid sustained Ca(2+) elevations in a limited number of cells and delayed secondary responses in neighbouring cells, seen as Ca(2+) waves whose propagation was consistent with extracellular diffusion of released ATP. Our experiments revealed that a single >10% stretch was sufficient to initiate intercellular purinergic signalling in alveolar cells, which may contribute to the regulation of surfactant secretion and wound healing.
Authors:
Ryszard Grygorczyk; Kishio Furuya; Masahiro Sokabe
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Video-Audio Media     Date:  2012-12-17
Journal Detail:
Title:  The Journal of physiology     Volume:  591     ISSN:  1469-7793     ISO Abbreviation:  J. Physiol. (Lond.)     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-03-04     Completed Date:  2013-08-21     Revised Date:  2014-03-07    
Medline Journal Info:
Nlm Unique ID:  0266262     Medline TA:  J Physiol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1195-215     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphate / metabolism*
Autocrine Communication* / drug effects
Calcium Signaling
Cell Line, Tumor
Cell Proliferation
Diffusion
Genes, Reporter
Humans
Image Processing, Computer-Assisted
Infrared Rays
Luminescent Measurements
Mechanotransduction, Cellular* / drug effects
Microscopy, Fluorescence
Paracrine Communication* / drug effects
Pneumocytes / drug effects,  metabolism*,  pathology
Pulmonary Stretch Receptors / metabolism*
Pulmonary Surfactant-Associated Proteins / metabolism
Purinergic P2 Receptor Antagonists / pharmacology
Receptors, Purinergic P2 / drug effects,  metabolism
Time Factors
Transfection
Wound Healing
Grant Support
ID/Acronym/Agency:
//Canadian Institutes of Health Research
Chemical
Reg. No./Substance:
0/Pulmonary Surfactant-Associated Proteins; 0/Purinergic P2 Receptor Antagonists; 0/Receptors, Purinergic P2; 8L70Q75FXE/Adenosine Triphosphate
Comments/Corrections

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