Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Mechanically induced ATP release from human osteoblastic cells.

M Romanello1, B Pani, M Bicego

  • 1Dipartimento di Biochimica, Biofisica, e Chimica delle Macromolecole, Università di Trieste, via Licio Giorgieri 1, Trieste, I-34127, Italy.

Biochemical and Biophysical Research Communications
|December 14, 2001
PubMed
Summary

Extracellular ATP release in HOBIT osteoblastic cells is mechanosensitive but not via connexin hemichannels. Mechanical stimuli enhance ATP efflux, which triggers intracellular calcium signals, indicating a physiological response.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The impact of intra-operative factors in otosclerosis outcomes: retrospective study in a tertiary centre.

Acta otorhinolaryngologica Italica : organo ufficiale della Societa italiana di otorinolaringologia e chirurgia cervico-facciale·2019
Same author

Connexin32 in the Peripheral Nervous System: Functional Analysis of Mutations Associated with X-linked Charcot-Marie-Tooth Syndrome and Implications for the Pathophysiology of the Disease.

Annals of the New York Academy of Sciences·2017
Same author

The strategies to reduce iron deficiency in blood donors randomized trial: design, enrolment and early retention.

Vox sanguinis·2014
Same author

Role of inflammation in non-allergic rhinitis.

Rhinology·2014
Same author

The child-rearing attitudes of the parents of (male and female) schizophrenics.

Indian journal of psychiatry·2011
Same author

Rehabilitative treatment in peripheral artery disease: protocol application and follow-up.

Minerva cardioangiologica·2010

Area of Science:

  • Cell Biology
  • Biochemistry
  • Physiology

Background:

  • Extracellular adenosine triphosphate (ATP) acts as a crucial autocrine/paracrine signaling molecule.
  • Cellular release of ATP is often mechanosensitive, yet the underlying molecular mechanisms remain largely undefined.
  • Proposed mechanisms include vesicular release, conductive channels, and ATP-binding cassette (ABC) transporters.

Purpose of the Study:

  • To investigate the molecular mechanism of extracellular ATP release in human HOBIT osteoblastic cells.
  • To determine the role of connexin hemichannels in ATP release under basal and stimulated conditions.
  • To elucidate the physiological consequences of mechanically induced ATP release.

Main Methods:

  • Luciferin-luciferase bioluminescence assay to quantify extracellular ATP.

Related Experiment Videos

  • Mechanical stimulation via medium displacement and hypotonic stress.
  • HOBIT cells were transfected to overexpress connexin43.
  • Treatment with cytochalasin D and manipulation of cyclic adenosine monophosphate (cAMP) levels.
  • Measurement of intracellular calcium (Ca2+) transients using fura-2 fluorescence.

    • Main Results:

    • Connexin hemichannels, specifically connexin43, were not significantly involved in ATP release in HOBIT cells.
    • Mechanical stimulation (medium displacement, hypotonic stress) significantly increased ATP efflux.
    • Cytochalasin D did not affect basal or stimulated ATP release.
    • Elevated cAMP levels consistently reduced ATP efflux under both basal and stimulated conditions.
    • Released ATP, induced by mechanical stimuli, elicited intracellular Ca2+ transients, confirming a physiological response.

    Conclusions:

    • Connexin hemichannels are not the primary pathway for mechanical ATP release in HOBIT osteoblastic cells.
    • Mechanical stress triggers significant ATP release in these cells, independent of actin cytoskeleton integrity (cytochalasin D).
    • Modulation of intracellular cAMP levels can regulate extracellular ATP release.
    • Mechanically induced extracellular ATP release activates downstream intracellular calcium signaling pathways in HOBIT cells.