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 language | English |
|---|---|
| Pages (from-to) | 283-292 |
| Number of pages | 10 |
| Journal | Free Radical Research |
| Volume | 28 |
| Issue number | 3 |
| State | Published - 1998 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
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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 journal › Article
}
TY - JOUR
T1 - Mechanism of peroxide-induced potentiation of sympathetic neurotransmission in bovine irides
T2 - Role of extracellular calcium
AU - Opere, Catherine A.
AU - Opere, Danish
AU - Ohia, Sunny E.
PY - 1998
Y1 - 1998
N2 - 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.
AB - 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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M3 - Article
VL - 28
SP - 283
EP - 292
JO - Free Radical Research
JF - Free Radical Research
SN - 1071-5762
IS - 3
ER -