Mechanism of peroxide-induced potentiation of sympathetic neurotransmission in bovine irides

Role of extracellular calcium

Catherine A. Opere, Danish Opere, Sunny E. Ohia

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Hydrogen peroxide (H2O2) and enzymes that regulate its metabolism are present in tissues of the anterior segment of the eye. We have previously shown that in vitro, H2O2 can enhance sympathetic neurotransmission in irides from several mammalian species. In the present study, we investigated the role of extracellular calcium in H2O2-induced potentiation of sympathetic neurotransmission in the bovine isolated iris. Isolated bovine hemiirides were incubated in a bicarbonate-buffered, carbogen-gassed Krebs buffer solution containing [3H]-norepinephrine ([3H]NE) for 60 min. After incubation, tissues were prepared for studies of [3H]NE release using the superfusion method. Release of [3H]NE was elicited by consecutive trains of electrical field stimulation. Removal of calcium from the buffer solution attenuated field-stimulated [3H]NE overflow in isolated, superfused bovine irides without affecting basal tritium efflux. H2O2 (1 mM) enhanced evoked [3H]NE release to the same extent in tissues exposed to buffer solutions containing normal calcium (1.3 mM) as in those containing low calcium (0.13 mM) or zero calcium. However, in the presence of zero-calcium buffer solution containing the chelator, EDTA (1 mM), H2O2 (1 mM) caused a gradual and sustained increase in basal tritium efflux. In buffer solutions containing high calcium (1.95 mM), the magnitude of H2O2-induced increase in field-stimulated [3H]NE release was significantly (P <0.05) attenuated. Although the neuronal calcium channel antagonist ω-conotoxin (20 nM) inhibited [3H]NE by 25%, it had no effect on H2O2 (1 mM)-induced potentiation of evoked [3H]NE overflow. We conclude that while trace amounts of extracellular calcium are necessary for H2O2-induced enhancement of sympathetic neurotransmission, increasing extracellular (buffer) calcium concentration impaired peroxide-induced enhancement of [3H]NE release. Furthermore, voltage-activated calcium channels may not be directly involved in peroxide-induced alteration of adrenergic neurosecretion in bovine isolated irides.

Original languageEnglish
Pages (from-to)283-292
Number of pages10
JournalFree Radical Research
Volume28
Issue number3
StatePublished - 1998

Fingerprint

Peroxides
Iris
Synaptic Transmission
Calcium
Buffers
Tritium
Tissue
Conotoxins
Neurosecretion
Anterior Eye Segment
Calcium Channel Blockers
Calcium Channels
Chelating Agents
Bicarbonates
Metabolism
Edetic Acid
Adrenergic Agents
Hydrogen Peroxide
Electric Stimulation
Norepinephrine

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Mechanism of peroxide-induced potentiation of sympathetic neurotransmission in bovine irides : Role of extracellular calcium. / Opere, Catherine A.; Opere, Danish; Ohia, Sunny E.

In: Free Radical Research, Vol. 28, No. 3, 1998, p. 283-292.

Research output: Contribution to journalArticle

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abstract = "Hydrogen peroxide (H2O2) and enzymes that regulate its metabolism are present in tissues of the anterior segment of the eye. We have previously shown that in vitro, H2O2 can enhance sympathetic neurotransmission in irides from several mammalian species. In the present study, we investigated the role of extracellular calcium in H2O2-induced potentiation of sympathetic neurotransmission in the bovine isolated iris. Isolated bovine hemiirides were incubated in a bicarbonate-buffered, carbogen-gassed Krebs buffer solution containing [3H]-norepinephrine ([3H]NE) for 60 min. After incubation, tissues were prepared for studies of [3H]NE release using the superfusion method. Release of [3H]NE was elicited by consecutive trains of electrical field stimulation. Removal of calcium from the buffer solution attenuated field-stimulated [3H]NE overflow in isolated, superfused bovine irides without affecting basal tritium efflux. H2O2 (1 mM) enhanced evoked [3H]NE release to the same extent in tissues exposed to buffer solutions containing normal calcium (1.3 mM) as in those containing low calcium (0.13 mM) or zero calcium. However, in the presence of zero-calcium buffer solution containing the chelator, EDTA (1 mM), H2O2 (1 mM) caused a gradual and sustained increase in basal tritium efflux. In buffer solutions containing high calcium (1.95 mM), the magnitude of H2O2-induced increase in field-stimulated [3H]NE release was significantly (P <0.05) attenuated. Although the neuronal calcium channel antagonist ω-conotoxin (20 nM) inhibited [3H]NE by 25{\%}, it had no effect on H2O2 (1 mM)-induced potentiation of evoked [3H]NE overflow. We conclude that while trace amounts of extracellular calcium are necessary for H2O2-induced enhancement of sympathetic neurotransmission, increasing extracellular (buffer) calcium concentration impaired peroxide-induced enhancement of [3H]NE release. Furthermore, voltage-activated calcium channels may not be directly involved in peroxide-induced alteration of adrenergic neurosecretion in bovine isolated irides.",
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