Modeling ischemia in vitro: Selective depletion of adenine and guanine nucleotide pools

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Intracellular ATP depletion is a hallmark event in ischemic injury. It has been extensively characterized in models of chemical anoxia in vitro. In contrast, the fate of GTP during ischemia remains unknown. We used LLC-PK proximal tubular cells to measure GTP and ATP changes during anoxia. In 45 min, antimycin A decreased ATP and GTP to 8% and 2% of controls, respectively. Ischemia in vivo resulted in comparable reductions in GTP and ATP. After 2 h of recovery, GTP levels in LLC-PK cells increased to 65% while ATP increased to 29%. We also investigated steady-state models of selective ATP or GTP depletion. Combinations of antimycin A and mycophenolic acid selectively reduced GTP to 51% or 25% of control. Similarly, alanosine selectively reduced ATP to 61% or 26% of control. Selective GTP depletion resulted in significant apoptosis. Selective ATP depletion caused mostly necrosis. These models of ATP or GTP depletion can prove useful in dissecting the relative contribution of the two nucleotides to the ischemic phenotype.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume279
Issue number4 48-4
StatePublished - 2000

Fingerprint

Guanine Nucleotides
Adenine Nucleotides
Guanosine Triphosphate
Ischemia
Adenosine Triphosphate
Antimycin A
alanosine
Mycophenolic Acid
Chemical Models
In Vitro Techniques
Necrosis
Nucleotides
Apoptosis
Phenotype
Recovery
Wounds and Injuries

Keywords

  • Alanosine
  • Apoptosis
  • ATP depletion
  • GTP depletion
  • Mycophenolic acid

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

@article{449950777245446bbe17bf12cc17fff9,
title = "Modeling ischemia in vitro: Selective depletion of adenine and guanine nucleotide pools",
abstract = "Intracellular ATP depletion is a hallmark event in ischemic injury. It has been extensively characterized in models of chemical anoxia in vitro. In contrast, the fate of GTP during ischemia remains unknown. We used LLC-PK proximal tubular cells to measure GTP and ATP changes during anoxia. In 45 min, antimycin A decreased ATP and GTP to 8{\%} and 2{\%} of controls, respectively. Ischemia in vivo resulted in comparable reductions in GTP and ATP. After 2 h of recovery, GTP levels in LLC-PK cells increased to 65{\%} while ATP increased to 29{\%}. We also investigated steady-state models of selective ATP or GTP depletion. Combinations of antimycin A and mycophenolic acid selectively reduced GTP to 51{\%} or 25{\%} of control. Similarly, alanosine selectively reduced ATP to 61{\%} or 26{\%} of control. Selective GTP depletion resulted in significant apoptosis. Selective ATP depletion caused mostly necrosis. These models of ATP or GTP depletion can prove useful in dissecting the relative contribution of the two nucleotides to the ischemic phenotype.",
keywords = "Alanosine, Apoptosis, ATP depletion, GTP depletion, Mycophenolic acid",
author = "Pierre Dagher",
year = "2000",
language = "English",
volume = "279",
journal = "American Journal of Physiology",
issn = "0193-1857",
publisher = "American Physiological Society",
number = "4 48-4",

}

TY - JOUR

T1 - Modeling ischemia in vitro

T2 - Selective depletion of adenine and guanine nucleotide pools

AU - Dagher, Pierre

PY - 2000

Y1 - 2000

N2 - Intracellular ATP depletion is a hallmark event in ischemic injury. It has been extensively characterized in models of chemical anoxia in vitro. In contrast, the fate of GTP during ischemia remains unknown. We used LLC-PK proximal tubular cells to measure GTP and ATP changes during anoxia. In 45 min, antimycin A decreased ATP and GTP to 8% and 2% of controls, respectively. Ischemia in vivo resulted in comparable reductions in GTP and ATP. After 2 h of recovery, GTP levels in LLC-PK cells increased to 65% while ATP increased to 29%. We also investigated steady-state models of selective ATP or GTP depletion. Combinations of antimycin A and mycophenolic acid selectively reduced GTP to 51% or 25% of control. Similarly, alanosine selectively reduced ATP to 61% or 26% of control. Selective GTP depletion resulted in significant apoptosis. Selective ATP depletion caused mostly necrosis. These models of ATP or GTP depletion can prove useful in dissecting the relative contribution of the two nucleotides to the ischemic phenotype.

AB - Intracellular ATP depletion is a hallmark event in ischemic injury. It has been extensively characterized in models of chemical anoxia in vitro. In contrast, the fate of GTP during ischemia remains unknown. We used LLC-PK proximal tubular cells to measure GTP and ATP changes during anoxia. In 45 min, antimycin A decreased ATP and GTP to 8% and 2% of controls, respectively. Ischemia in vivo resulted in comparable reductions in GTP and ATP. After 2 h of recovery, GTP levels in LLC-PK cells increased to 65% while ATP increased to 29%. We also investigated steady-state models of selective ATP or GTP depletion. Combinations of antimycin A and mycophenolic acid selectively reduced GTP to 51% or 25% of control. Similarly, alanosine selectively reduced ATP to 61% or 26% of control. Selective GTP depletion resulted in significant apoptosis. Selective ATP depletion caused mostly necrosis. These models of ATP or GTP depletion can prove useful in dissecting the relative contribution of the two nucleotides to the ischemic phenotype.

KW - Alanosine

KW - Apoptosis

KW - ATP depletion

KW - GTP depletion

KW - Mycophenolic acid

UR - http://www.scopus.com/inward/record.url?scp=0033679661&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033679661&partnerID=8YFLogxK

M3 - Article

C2 - 11003607

AN - SCOPUS:0033679661

VL - 279

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0193-1857

IS - 4 48-4

ER -