Changes of nucleotide patterns in liver, muscle and blood during the growth of Ehrlich ascites cells: application of the reversed-phase and ion-pair reversed-phase high-performance liquid chromatography with radial compression column

Grune, Tilman; Siems, Werner; Gerber, Gerhard; Tikhonov, Yuri V.; Pimenov, A M; Toguzov, R T

doi:10.1016/0378-4347(91)80276-i

Cited by 23 publications

(4 citation statements)

References 8 publications

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“…Because CMP is likely to increase during anoxia, this could lead to overestimation of GTP. In addition, nucleotides are notoriously difficult to resolve with reverse-phase protocols in the absence of ion pairing (10). Furthermore, perchloric acid extraction can interfere with peak resolution due to residual perchlorate ions, a problem avoided by using acetonitrile extraction (1,29).…”

Section: Discussionmentioning

confidence: 99%

“…The column used was a 4-m Nova-Pak C 18 cartridge (100 mm by 8 mm ID), equipped with a radial compression chamber (Waters, Milford, MA). The buffer consisted of 20% acetonitrile, 10 mM ammonium phosphate, and 2 mM PIC Reagent A ion-pairing reagent (Waters) and was run isocratically at 2 ml/min (10). Samples were diluted in half, and the injection volume was 100 l. A HP Chemstation model 1100 was used (Hewlett-Packard, Wilmington, DE), and the ultraviolet (UV) detector was set at 254 nm.…”

Section: Methodsmentioning

confidence: 99%

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

Dagher

2000

American Journal of Physiology-Cell Physiology

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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

Dagher

2000

American Journal of Physiology-Cell Physiology

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“…The column used to perform the HPLC analysis was a 4 mm Nova-Pack C 18 cartridge (100 Â 8 mm inner diameter) equipped with a radial compression chamber (Waters, Milford, MA, USA). The buffer consisted of 20% acetonitrile, 10 mM ammonium phosphate, and 2 mM PIC-A ion-pairing reagent (Waters) and was run isocratically at 1 min/ml (40). Waters HPLC model 510 was used (Waters, Milford, MA, USA), and the ultraviolet (UV) detector model 440 (Waters) was set at 254 nm.…”

Section: Nucleotide Extraction and Hplc Analysismentioning

confidence: 99%

Specific progressive cAMP reduction implicates energy deficit in presymptomatic Huntington's disease knock-in mice

Ginés

Seong

Fossale

et al. 2003

Human Molecular Genetics

253

130

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Defects in gene transcription and mitochondrial function have been implicated in the dominant disease process that leads to the loss of striatal neurons in Huntington's disease (HD). Here we have used precise genetic HD mouse and striatal cell models to investigate the hypothesis that decreased cAMP responsive element (CRE)-mediated gene transcription may reflect impaired energy metabolism. We found that reduced CRE-signaling in Hdh(Q111) striatum, monitored by brain derived neurotrophic factor and phospho-CRE binding protein (CREB), predated inclusion formation. Furthermore, cAMP levels in Hdh(Q111) striatum declined from an early age (10 weeks), and cAMP was significantly decreased in HD postmortem brain and lymphoblastoid cells, attesting to a chronic deficit in man. Reduced CRE-signaling in cultured STHdh(Q111) striatal cells was associated with cytosolic CREB binding protein that mirrored diminished cAMP synthesis. Moreover, mutant cells exhibited mitochondrial respiratory chain impairment, evidenced by decreased ATP and ATP/ADP ratio, impaired MTT conversion and heightened sensitivity to 3-nitropropionic acid. Thus, our findings strongly suggest that impaired ATP synthesis and diminished cAMP levels amplify the early HD disease cascade by decreasing CRE-regulated gene transcription and altering energy dependent processes essential to neuronal cell survival.

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“…Nucleotides were separated via HPLC by utilizing a 4-m Nova-Pack C18 cartridge (100 mm by 8 mm inner diameter) equipped with a radial compression chamber (Waters, Millford, MA). The buffer consisted of 20% acetonitrile, 10 mM ammonium phosphate, and 2 mM PIC-A ion pairing reagent (Waters) and was run isocratically at 2 ml/min (12). Samples were diluted by onehalf, and the injection volume was 100 l. An HP Chemstation model no.…”

Section: Reagents and Antibodiesmentioning

confidence: 99%

Rho-kinase regulates myosin II activation in MDCK cells during recovery after ATP depletion

Sutton

Mang

Atkinson

2001

American Journal of Physiology-Renal Physiology

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Alterations in the actin cytoskeleton of renal tubular epithelial cells during periods of ischemic injury and recovery have important consequences for normal cell and kidney function. Myosin II has been demonstrated to be an important effector in organizing basal actin structures in some cell types. ATP depletion in vitro has been demonstrated to recapitulate alterations of the actin cytoskeleton in renal tubular epithelial cells observed during renal ischemia in vivo. We utilized this reversible cell culture model of ischemia to examine the correlation of the activation state and cellular distribution of myosin II with disruption of actin stress fibers in Madin-Darby canine kidney (MDCK) cells during ATP depletion and recovery from ATP depletion. We found that myosin II inactivation occurs rapidly and precedes dissociation of myosin II from actin stress fibers during ATP depletion. Myosin II activation temporally correlates with colocalization of myosin II to reorganizing stress fibers during recovery from ATP depletion. Furthermore, myosin activation and actin stress fiber formation were found to be Rho-associated Ser/Thr protein kinase dependent during recovery from ATP depletion.

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Changes of nucleotide patterns in liver, muscle and blood during the growth of Ehrlich ascites cells: application of the reversed-phase and ion-pair reversed-phase high-performance liquid chromatography with radial compression column

Cited by 23 publications

References 8 publications

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

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

Specific progressive cAMP reduction implicates energy deficit in presymptomatic Huntington's disease knock-in mice

Rho-kinase regulates myosin II activation in MDCK cells during recovery after ATP depletion

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