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Characterization of Ca <sup>2+</sup> ‐Based Action Potentials in Horizontal Cells in the Goldfish ( <i>Carassius auratus</i> ) Retina

2019· article· en· W3176889704 on OpenAlexafffundabout
Michael William Country, Michael G. Jonz

Bibliographic record

VenueThe FASEB Journal · 2019
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicRetinal Development and Disorders
Canadian institutionsUniversity of Ottawa
FundersNatural Sciences and Engineering Research Council of Canada
KeywordsRyanodine receptorMembrane potentialRetinaBiophysicsDantroleneChemistryReceptor potentialIntracellularDepolarizationElectrophysiologyInhibitory postsynaptic potentialCalcium imagingAgonistCalciumBiologyReceptorEndocrinologyNeuroscienceBiochemistry

Abstract

fetched live from OpenAlex

About 65 years ago, the first intracellular recordings ever to be made in the retina were made in horizontal cells (HCs) of goldfish ( Carassius auratus ). These recordings were some of the first examples of graded potentials (where membrane potential changes in proportion with a stimulus), and it was assumed that outer retinal neurons did not present action potentials (APs). HCs are interneurons which provide inhibitory feedback to photoreceptors, and in vision they are responsible for improving contrast, edge detection, and color opponency. Since these first recordings, Ca 2+ ‐based APs and spontaneous increases in intracellular Ca 2+ concentration ([Ca 2+ ] i ) have been found in fish HCs, although they are poorly understood and their role in vision (if any) is unknown. We characterized spontaneous, Ca 2+ ‐based APs in isolated goldfish HCs by measuring changes in [Ca 2+ ] i and membrane potential ( V m ). Using Fura‐2 Ca 2+ imaging, we found spontaneous Ca 2+ activity characterized by transient elevation in [Ca 2+ ] i of short (seconds) and long (minutes) duration, in 157/177 cells (89%). These APs were similar in amplitude to Ca 2+ responses evoked by glutamate. APs were reversibly eliminated in Ca 2+ ‐free solutions (n=18), and were abolished by the L‐type Ca 2+ channel blocker, nifedipine (100 μM; n=5). APs were also dependent on intracellular Ca 2+ stores: they were abolished by the ryanodine receptor antagonists, ryanodine (20 μM; in 7/8 cells) and dantrolene (50 μM; n=7). The ryanodine receptor agonist, caffeine (n=10), increased AP frequency 2.8‐fold (p<0.0002) and reduced median duration (29.3 s to 18.7 s), time to peak (12.3 s to 7.8 s), and area under the curve (by 59.6%). When caffeine was co‐applied with the store‐operated channel antagonist, 2‐aminoethyldiphenyl borate (2‐APB), frequency was unaffected, further confirming a role for stores. We tracked changes in V m with whole‐cell current‐clamp recording, and with the voltage‐sensitive dye, FluoVolt. Phenotypes of APs in FluoVolt (n=6) and current‐clamp (48/57 cells; 84%) experiments displayed a sharp rise to peak, followed by a slow decline for the duration of the transient, and a steep return to baseline. In current‐clamp experiments, APs were dependent on L‐type Ca 2+ channel activity. APs were blocked by Co 2+ (5 mM; n=5) and nifedipine (100 μM; n=5), and were amplified and prolonged by the L‐type channel‐permeant ion, Ba 2+ (15 mM; n=6). In addition, APs were abolished in 6/9 cells in Ca 2+ ‐free solution. Collectively, our data suggest that activation of voltage‐dependent L‐type Ca 2+ channels leads to Ca 2+ influx, depolarization of V m , and additional Ca 2+ release from stores via ryanodine receptors. Understanding this phenomenon is a first step to elucidating a possible role for APs in vision or retinal physiology. This work also challenges the long‐held belief that outer retinal neurons do not present APs, and may lead to new paradigms about how visual information is encoded in the vertebrate retina. Support or Funding Information We acknowledge the support of the Natural Sciences and Engineering Research Council of Canada (NSERC), grant 342303. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

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How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.002
Threshold uncertainty score0.004

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.007
GPT teacher head0.218
Teacher spread0.211 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations0
Published2019
Admission routes3
Has abstractyes

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