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The Journal of Physiology
Article . 2002 . Peer-reviewed
License: Wiley Online Library User Agreement
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Time course and magnitude of the calcium release induced by bright light in salamander rods

Authors: Hugh R, Matthews; Gordon L, Fain;

Time course and magnitude of the calcium release induced by bright light in salamander rods

Abstract

Changes in fluorescence were measured with an argon ion laser from the outer segments of isolated salamander rods containing the Ca2+‐sensitive fluorescent dye fluo‐5F. When the outer segments were exposed to a 0Ca2+/0Na+ solution designed to minimise surface membrane Ca2+ fluxes, exposure to intense light from the laser evoked a slow increase in fluorescence, reflecting a light‐induced rise in outer segment [Ca2+]i. The time course of this slow fluorescence rise could be fitted with the sum of two asymptotic exponential functions of approximately equal amplitude, having time constants of approximately 200 ms and 5.7 s. When rods were exposed to saturating background light to reduce outer segment [Ca2+]i before laser illumination, the relative amplitude of the two exponentials was altered so as to reduce the contribution from the one with the shorter time constant. Examination of the initial time course of fluorescence when recording at high temporal resolution revealed a further rapid rise with a time constant of 1–2 ms, which could be observed even from rods in Ringer solution. This initial rapid rise could be abolished by pre‐exposing the rod to bleaching illumination, whether the bleach was given in Ringer solution or in 0Ca2+/0Na+ solution. It would therefore appear that the rapid rise in fluorescence is generated in some way by the bleaching of the photopigment. Unlike the slower components of fluorescence increase, the rapid initial rise was virtually unaffected in waveform or amplitude when rods were pre‐exposed in Ringer solution to light which was bright enough to suppress completely the circulating current but which bleached a negligible fraction of the photopigment. Furthermore, pre‐incubation with the AM ester of the Ca2+ chelator BAPTA, although completely abolishing the slower components of fluorescence increase, had virtually no effect on the rapid rise. These results indicate that the rapid component, though triggered by rhodopsin bleaching, does not reflect an increase in outer segment [Ca2+]i. Neither the rapid nor the slower components of fluorescence increase were affected by exposure of the outer segment to 10 μm of the membrane‐permeant compound N,N,N′,N′‐tetrakis(2‐pyridyl‐methyl)ethylenediamine (TPEN), which chelates heavy metals such as Zn2+, or 100 μm 2‐aminoethoxydiphenylborate (2‐APB), a membrane‐permeant blocker of IP3 receptors. These results appear to exclude a role for changes in heavy metal concentration or Ca2+ release via IP3 receptors in the light‐induced increases in dye fluorescence. Estimates of absolute Ca2+ concentration and of rod buffering capacity suggest that the slower components of fluorescence increase represent the release of around 10–50 μmoles Ca2+ per litre cytoplasmic volume from bound or sequestered stores after bleaching.

Related Organizations
Keywords

Boron Compounds, Time Factors, Light, In Vitro Techniques, Ethylenediamines, Ambystoma, Fluorescence, Retinal Rod Photoreceptor Cells, Animals, Calcium, Egtazic Acid, Chelating Agents

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
15
Average
Top 10%
Top 10%
bronze