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Purinergic receptors and regulatory volume decrease in seabream (Sparus aurata) hepatocytes: a videometric study.

Agata Torre1, Francesca Trischitta2, Caterina Faggio1

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Fish Physiology and Biochemistry
|May 16, 2012
PubMed
Summary

Isolated fish liver cells (hepatocytes) can regulate their volume (RVD) under hypotonic stress. Extracellular adenosine triphosphate (ATP) plays a key role in this process, acting via P2 receptors.

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Area of Science:

  • Cellular Physiology
  • Marine Biology
  • Biochemistry

Background:

  • Hepatocytes of Sparus aurata exhibit regulatory volume decrease (RVD) in response to hypotonic stress.
  • Previous studies indicated the cell's capacity for RVD, but the underlying mechanisms remain incompletely understood.
  • The role of adenosine triphosphate (ATP) in cellular volume regulation requires further investigation.

Purpose of the Study:

  • To investigate the involvement of extracellular adenosine triphosphate (ATP) in the regulatory volume decrease (RVD) of Sparus aurata hepatocytes.
  • To elucidate the specific purinergic receptors (P1 and P2) mediating ATP's effects on RVD.
  • To identify potential mechanisms of regulated ATP release from hepatocytes.

Main Methods:

  • Videometric analysis of isolated Sparus aurata hepatocytes under hypotonic conditions.
  • Pharmacological manipulation using ATP scavengers (apyrase), receptor agonists (ATPγS, adenosine), and antagonists (suramin, 8-PT).
  • Assessment of ATP release mechanisms using specific inhibitors, including verapamil.

Main Results:

  • Apyrase significantly inhibited the RVD response, confirming the involvement of extracellular ATP.
  • ATPγS (P2 receptor agonist) stimulated RVD, while suramin (P2 antagonist) inhibited it, indicating P2 receptor mediation.
  • Adenosine (P1 receptor agonist) and 8-PT (P1 antagonist) showed opposing effects, suggesting complex P1 receptor involvement.
  • ATP release appears to be verapamil-sensitive, implicating the P-glycoprotein transporter.

Conclusions:

  • Extracellular ATP is crucial for the regulatory volume decrease (RVD) response in Sparus aurata hepatocytes.
  • The stimulatory effect of ATP on RVD is primarily mediated through P2 purinergic receptors.
  • The regulated release of ATP from these cells may involve the P-glycoprotein transporter.