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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

We present a behavioral chamber designed to assess cognitive performance. We provide data showing that once acquired, zebrafish remember the task 8 weeks later. We also show that hyperglycemic zebrafish have altered cognitive performance, indicating that this paradigm is applicable to studies assessing cognition and memory.

Abstract

Neurodegenerative diseases are age-dependent, debilitating, and incurable. Recent reports have also correlated hyperglycemia with changes in memory and/or cognitive impairment. We have modified and developed a three-chamber choice cognitive task similar to that used with rodents for use with hyperglycemic zebrafish. The testing chamber consists of a centrally located starting chamber and two choice compartments on either side, with a shoal of conspecifics used as the reward. We provide data showing that once acquired, zebrafish remember the task at least 8 weeks later. Our data indicate that zebrafish respond robustly to this reward, and we have identified cognitive deficits in hyperglycemic fish after 4 weeks of treatment. This behavioral assay may also be applicable to other studies related to cognition and memory.

Introduction

Neurodegenerative diseases are age-dependent, debilitating, and incurable. These diseases are increasing in prevalence, resulting in an urgent need to improve upon and develop new therapeutic strategies. The onset and presentation of each disease is unique, as some affect language, motor, and autonomic brain regions, while others cause learning deficits and memory loss1. Most notably, cognitive deficits and/or impairment are the most prevalent complications across all neurodegenerative diseases2. In hopes of shedding light on the underlying mechanisms involved in these neurodegenerative diseases, the use of many differen....

Protocol

All experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC) at American University (protocol # 1606, 19-02).

1. Animals

  1. Animal rearing and maintenance
    1. Obtain adult wild-type zebrafish (Danio rerio) aged 4–11 months as embryos and rear them in-house.
    2. Maintain the fish in an aquatic rack system at 28–29 °C on a 14-h light:10-h dark photoperiod.
    3. Feed the fish twice per day with commerc.......

Representative Results

Acclimation to the behavior chamber involves three days of training: 2 days of group acclimation followed by 1 day of individual acclimation. However, because we could not distinguish individual zebrafish from one another, we were only able to collect data during individual acclimation. At this time, experimental animals (n = 30), conditioned using a shoal-based reward, took an average of 125.11 s to reach their first decision (Figure 2A) and an average of 725.34 s (12 min) to complete the e.......

Discussion

Although there has been tremendous growth in the amount and variety of neuroscience research performed using zebrafish in the past 15 years24, behavioral assays are lacking in this species compared to mammalian model systems11,25,26. Here, we show that a three-chamber choice task developed for use with rodents can be adapted to assess the acquisition and reversal of a visual discrimination learning in zeb.......

Acknowledgements

We thank Sabrina Jones for her assistance adapting a rodent three-chamber choice paradigm to the zebrafish model and Jeremy Popowitz and Allison Murk for their help on behavior collection days, assistance with running trials, animal care, and tank set-up. Special thanks also to James M. Forbes (Mechanical Engineer) for his assistance with the 3-chamber choice tank design and construction.

Funding: VPC and TLD received a joint Faculty Research Support grant (FRSG) from American University College of Arts and Sciences. CJR received support from American University College of Arts and Sciences Graduate Student Support.

....

Materials

NameCompanyCatalog NumberComments
Champion Sports Stopwatch Timer Set: Waterproof, Handheld Digital Clock Sport Stopwatches with Large Display for Kids or Coach - Bright Colored 6 PackAmazonN/Ahttps://www.amazon.com/Champion-Sports-910SET-Stopwatch-Timer/dp/B001CD9LJK/ref=sr_1_17?dchild=1&keywords=stopwatch+for+sports&qid=1597081570&sr=8-17
Recommend two of different colors; one for choice latency and one for time to completion
Coofficer Extra Large Binder Clips 2-Inch (24 Pack), Big Paper Clamps for Office Supplies, BlackAmazonN/Ahttps://www.amazon.com/Coofficer-Binder-2-Inch-Clamps-Supplies/dp/B07C94YCR5/ref=sr_1_3_sspa?dchild=1&keywords=large+binder+clips&qid=1597081521&sr=8-3-spons&psc=1&spLa=ZW5jcnlwdGVkUXVhbGlmaWVyPUExUENWUTRZVjlIWEVPJmVuY3J5cHRlZElkPUEwNDQ5NDU0MlpSREkwTFlLSThVQiZlbmNyeXB0ZWRBZElkPUEwMTg5NDI3MllRV1EzOUdWTVpSOCZ3aWRnZXROYW1lPXNwX2F0ZiZhY3Rpb249Y2xpY2tSZWRpcmVjdCZkb05vdExvZ0NsaWNrPXRydWU=
Marineland® Silicone Aquarium SealantPetsmartItem #2431002
PVC (Polyvinyl Chloride) Sheet, Opaque Gray, Standard Tolerance, UL 94/ASTM D1784, 0.125" Thickness, 12" Width, 24" LengthAmazonN/Ahttps://www.amazon.com/Polyvinyl-Chloride-Standard-Tolerance-Thickness/dp/B000MAMGEQ/ref=sr_1_2?dchild=1&keywords=grey+PVC+sheet&qid=1597081440&sr=8-2
Steelworks 1/4-in W x 8-ft L Mill Finished Aluminum Weldable Trim U-shaped ChannelLowesItem #55979Model #11377https://www.lowes.com/pd/Steelworks-1-4-in-W-x-8-ft-L-Mill-Finished-Aluminum-Weldable-Trim-Channel/3058181
Tetra 10 Gallon Fish tankPetsmartItem #5271256
Top Fin Fine Mesh Fish Net (3 in)PetsmartItem #5175115

References

  1. Gitler, A. D., Dhillon, P., Shorter, J. Neurodegenerative disease: models, mechanisms, and a new hope. Disease Models & Mechanisms. 10, 499-502 (2017).
  2. Perry, R. J., Watson, P., Hodges, J. R.

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