Real-time imaging of ATP release induced by mechanical stretch in human airway smooth muscle cells

Norihiro Takahara, Satoru Ito, Kishio Furuya, Keiji Naruse, Hiromichi Aso, Masashi Kondo, Masahiro Sokabe, Yoshinori Hasegawa

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Airway smooth muscle (ASM) cells within the airway walls are continually exposed to mechanical stimuli, and exhibit various functions in response to these mechanical stresses. ATP acts as an extracellular mediator in the airway. Moreover, extracellular ATP is considered to play an important role in the pathophysiology of asthma and chronic obstructive pulmonary disease. However, it is not known whether ASM cells are cellular sources of ATP secretion in the airway. We therefore investigated whether mechanical stretch induces ATP release from ASM cells. Mechanical stretch was applied to primary human ASM cells cultured on a silicone chamber coated with type I collagen using a stretching apparatus. Concentrations of ATP in cell culture supernatants measured by luciferin-luciferase bioluminescence were significantly elevated by cyclic stretch (12 and 20% strain). We further visualized the stretch-induced ATP release from the cells in real time using a luminescence imaging system, while acquiring differential interference contrast cell images with infrared optics. Immediately after a single uniaxial stretch for 1 second, strong ATP signals were produced by a certain population of cells and spread to surrounding spaces. The cyclic stretch-induced ATP release was significantly reduced by inhibitors of Ca2+-dependent vesicular exocytosis, 1,2-bis(o-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid tetraacetoxymethyl ester, monensin, N-ethylmaleimide, and bafilomycin. In contrast, the stretch-induced ATP release was not inhibited by a hemichannel blocker, carbenoxolone, or blockade of transient receptor potential vanilloid 4 by short interfering RNA transfection or ruthenium red. These findings reveal a novel property of ASM cells: mechanically induced ATP release may be a cellular source of ATP in the airway.

Original languageEnglish
Pages (from-to)772-782
Number of pages11
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume51
Issue number6
DOIs
Publication statusPublished - 01-12-2014
Externally publishedYes

Fingerprint

Smooth Muscle Myocytes
Muscle
Adenosine Triphosphate
Cells
Imaging techniques
Carbenoxolone
Bioluminescence
TRPV Cation Channels
Ruthenium Red
Monensin
Pulmonary diseases
Mechanical Stress
Ethylmaleimide
Ethane
Exocytosis
Silicones
Collagen Type I
Luminescence
Luciferases
Cell culture

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Takahara, Norihiro ; Ito, Satoru ; Furuya, Kishio ; Naruse, Keiji ; Aso, Hiromichi ; Kondo, Masashi ; Sokabe, Masahiro ; Hasegawa, Yoshinori. / Real-time imaging of ATP release induced by mechanical stretch in human airway smooth muscle cells. In: American Journal of Respiratory Cell and Molecular Biology. 2014 ; Vol. 51, No. 6. pp. 772-782.
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Real-time imaging of ATP release induced by mechanical stretch in human airway smooth muscle cells. / Takahara, Norihiro; Ito, Satoru; Furuya, Kishio; Naruse, Keiji; Aso, Hiromichi; Kondo, Masashi; Sokabe, Masahiro; Hasegawa, Yoshinori.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 51, No. 6, 01.12.2014, p. 772-782.

Research output: Contribution to journalArticle

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