Effect of Hydrogen Peroxide on Amino Acid Concentrations in Bovine Retina and Vitreous Humor, Ex Vivo

A. M. LeDay, S. Ganguly, K. H. Kulk̃arni, Alekha K. Dash, Catherine A. Opere, Sunny E. Ohia

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In a previous study, we showed evidence that oxidative stress induced by hydrogen peroxide (H2O2) can inhibit the release of [ 3H]-D-aspartate from the bovine isolated retina, in vitro. The aim of the present study was to investigate the effect of H2O 2 on glutamate and glycine levels in the bovine retina and vitreous humor, ex vivo. Furthermore, we examined whether inhibition of catalase activity with 3-amino-triazole had any effect on the concentrations of these amino acids in the posterior segment of the bovine eye. Whole eye organ cultures were prepared by incubating tissues in oxygenated Krebs solution at 37 °C for 30 min. After incubation, H2O2 (1-100 μM) or sterile distilled water was injected intravitreally into each eye. Thirty minutes after injection, the retina and vitreous humor were removed for analysis of glutamate and glycine by high performance liquid chromatography (HPLC) with fluorescence detection. Exogenously applied H2O 2 (1-100 μM) caused a concentration-related decrease in both glutamate and glycine levels in the bovine retina. Furthermore, while H 2O2 (1-10 μM) caused a concentration-dependent decrease in glycine levels in the vitreous humor, it had no significant effect on glutamate levels. The catalase inhibitor, 3-amino-triazole (10 mM), caused a significant reduction in both glutamate and glycine levels in the bovine retina, ex vivo. Likewise, 3-AT caused an attenuation in both glutamate and glycine concentration in the vitreous humor. We conclude that oxidative stress induced by H2O2 can alter the release and/or availability of amino acids in the posterior segment of bovine eyes.

Original languageEnglish
Pages (from-to)695-701
Number of pages7
JournalMethods and Findings in Experimental and Clinical Pharmacology
Volume25
Issue number9
DOIs
StatePublished - Nov 2003

Fingerprint

Vitreous Body
Glycine
Hydrogen Peroxide
Retina
Glutamic Acid
Amino Acids
Posterior Eye Segment
Triazoles
Catalase
Oxidative Stress
D-Aspartic Acid
Organ Culture Techniques
Fluorescence
High Pressure Liquid Chromatography
Injections
Water

All Science Journal Classification (ASJC) codes

  • Pharmacology (medical)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Effect of Hydrogen Peroxide on Amino Acid Concentrations in Bovine Retina and Vitreous Humor, Ex Vivo. / LeDay, A. M.; Ganguly, S.; Kulk̃arni, K. H.; Dash, Alekha K.; Opere, Catherine A.; Ohia, Sunny E.

In: Methods and Findings in Experimental and Clinical Pharmacology, Vol. 25, No. 9, 11.2003, p. 695-701.

Research output: Contribution to journalArticle

@article{193d5628000e462a8ea6cf272daec818,
title = "Effect of Hydrogen Peroxide on Amino Acid Concentrations in Bovine Retina and Vitreous Humor, Ex Vivo",
abstract = "In a previous study, we showed evidence that oxidative stress induced by hydrogen peroxide (H2O2) can inhibit the release of [ 3H]-D-aspartate from the bovine isolated retina, in vitro. The aim of the present study was to investigate the effect of H2O 2 on glutamate and glycine levels in the bovine retina and vitreous humor, ex vivo. Furthermore, we examined whether inhibition of catalase activity with 3-amino-triazole had any effect on the concentrations of these amino acids in the posterior segment of the bovine eye. Whole eye organ cultures were prepared by incubating tissues in oxygenated Krebs solution at 37 °C for 30 min. After incubation, H2O2 (1-100 μM) or sterile distilled water was injected intravitreally into each eye. Thirty minutes after injection, the retina and vitreous humor were removed for analysis of glutamate and glycine by high performance liquid chromatography (HPLC) with fluorescence detection. Exogenously applied H2O 2 (1-100 μM) caused a concentration-related decrease in both glutamate and glycine levels in the bovine retina. Furthermore, while H 2O2 (1-10 μM) caused a concentration-dependent decrease in glycine levels in the vitreous humor, it had no significant effect on glutamate levels. The catalase inhibitor, 3-amino-triazole (10 mM), caused a significant reduction in both glutamate and glycine levels in the bovine retina, ex vivo. Likewise, 3-AT caused an attenuation in both glutamate and glycine concentration in the vitreous humor. We conclude that oxidative stress induced by H2O2 can alter the release and/or availability of amino acids in the posterior segment of bovine eyes.",
author = "LeDay, {A. M.} and S. Ganguly and Kulk̃arni, {K. H.} and Dash, {Alekha K.} and Opere, {Catherine A.} and Ohia, {Sunny E.}",
year = "2003",
month = "11",
doi = "10.1358/mf.2003.25.9.793336",
language = "English",
volume = "25",
pages = "695--701",
journal = "Methods and Findings in Experimental and Clinical Pharmacology",
issn = "0379-0355",
publisher = "Prous Science",
number = "9",

}

TY - JOUR

T1 - Effect of Hydrogen Peroxide on Amino Acid Concentrations in Bovine Retina and Vitreous Humor, Ex Vivo

AU - LeDay, A. M.

AU - Ganguly, S.

AU - Kulk̃arni, K. H.

AU - Dash, Alekha K.

AU - Opere, Catherine A.

AU - Ohia, Sunny E.

PY - 2003/11

Y1 - 2003/11

N2 - In a previous study, we showed evidence that oxidative stress induced by hydrogen peroxide (H2O2) can inhibit the release of [ 3H]-D-aspartate from the bovine isolated retina, in vitro. The aim of the present study was to investigate the effect of H2O 2 on glutamate and glycine levels in the bovine retina and vitreous humor, ex vivo. Furthermore, we examined whether inhibition of catalase activity with 3-amino-triazole had any effect on the concentrations of these amino acids in the posterior segment of the bovine eye. Whole eye organ cultures were prepared by incubating tissues in oxygenated Krebs solution at 37 °C for 30 min. After incubation, H2O2 (1-100 μM) or sterile distilled water was injected intravitreally into each eye. Thirty minutes after injection, the retina and vitreous humor were removed for analysis of glutamate and glycine by high performance liquid chromatography (HPLC) with fluorescence detection. Exogenously applied H2O 2 (1-100 μM) caused a concentration-related decrease in both glutamate and glycine levels in the bovine retina. Furthermore, while H 2O2 (1-10 μM) caused a concentration-dependent decrease in glycine levels in the vitreous humor, it had no significant effect on glutamate levels. The catalase inhibitor, 3-amino-triazole (10 mM), caused a significant reduction in both glutamate and glycine levels in the bovine retina, ex vivo. Likewise, 3-AT caused an attenuation in both glutamate and glycine concentration in the vitreous humor. We conclude that oxidative stress induced by H2O2 can alter the release and/or availability of amino acids in the posterior segment of bovine eyes.

AB - In a previous study, we showed evidence that oxidative stress induced by hydrogen peroxide (H2O2) can inhibit the release of [ 3H]-D-aspartate from the bovine isolated retina, in vitro. The aim of the present study was to investigate the effect of H2O 2 on glutamate and glycine levels in the bovine retina and vitreous humor, ex vivo. Furthermore, we examined whether inhibition of catalase activity with 3-amino-triazole had any effect on the concentrations of these amino acids in the posterior segment of the bovine eye. Whole eye organ cultures were prepared by incubating tissues in oxygenated Krebs solution at 37 °C for 30 min. After incubation, H2O2 (1-100 μM) or sterile distilled water was injected intravitreally into each eye. Thirty minutes after injection, the retina and vitreous humor were removed for analysis of glutamate and glycine by high performance liquid chromatography (HPLC) with fluorescence detection. Exogenously applied H2O 2 (1-100 μM) caused a concentration-related decrease in both glutamate and glycine levels in the bovine retina. Furthermore, while H 2O2 (1-10 μM) caused a concentration-dependent decrease in glycine levels in the vitreous humor, it had no significant effect on glutamate levels. The catalase inhibitor, 3-amino-triazole (10 mM), caused a significant reduction in both glutamate and glycine levels in the bovine retina, ex vivo. Likewise, 3-AT caused an attenuation in both glutamate and glycine concentration in the vitreous humor. We conclude that oxidative stress induced by H2O2 can alter the release and/or availability of amino acids in the posterior segment of bovine eyes.

UR - http://www.scopus.com/inward/record.url?scp=0347762833&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0347762833&partnerID=8YFLogxK

U2 - 10.1358/mf.2003.25.9.793336

DO - 10.1358/mf.2003.25.9.793336

M3 - Article

VL - 25

SP - 695

EP - 701

JO - Methods and Findings in Experimental and Clinical Pharmacology

JF - Methods and Findings in Experimental and Clinical Pharmacology

SN - 0379-0355

IS - 9

ER -