Name: Experimentell akvariekonfiguration och videoinspelningsinställning för beteendetestning hos vuxna zebrafiskar
Uploaded: 2023-11-03T12:40:00
Description: Watch this Scientific Journal Video about Experimental Tank Configuration and Video Recording Setup for Behavioral Testing in Adult Zebrafish at JoVE.com

experimental tank configuration and video recording setup for behavioral testing in adult zebrafish

Beteendetestbatteri för att bedöma neurotoxiska effekter hos vuxna zebrafiskar

The zebrafish (Danio rerio) is a popular model vertebrate species for ecotoxicology, drug discovery, and safety pharmacology studies. Its low cost, well-established molecular genetic tools, and conservation of key physiological processes involved in the morphogenesis and maintenance of the nervous system make zebrafish an ideal animal model for neuroscience research, including neurobehavioral toxicology1,2. The main endpoint for evaluating the neurotoxicity of a chemical was, until recently, the presence of neuropathological effects. Lately, however, behavioral endpoints have been incorporated into neurotoxicological screening protocols, and these functional outcomes are now commonly used to identify and determine the potential neurotoxicity of chemicals3,4. Moreover, behavioral endpoints are highly relevant from an ecological point of view, as even a very mild behavioral change in fish could endanger the survival of the animal in natural conditions5.
One of the most used behavioral assays in adult zebrafish research is the novel tank test (NTT), which measures anxiety-like behavior6,7. In this assay, fish are exposed to novelty (fish are placed in an unfamiliar tank), a mild aversive stimulus and their behavioral responses are observed. NTT is used to assess basal locomotor activity, geotaxis, freezing, and erratic movements of fish, principally. Erratic8 is characterized by abrupt changes of direction (zigzagging) and repeated episodes of accelerations (darting). It is an alarm reaction and is usually observed before or after freezing episodes. Freezing behavior corresponds to a complete cessation of the fish&#39;s movements (except for opercular and ocular movements) while on the bottom of the tank, as distinguished from immobility caused by sedation, which causes hypolocomotion, akinesia, and sinking8. Freezing is usually related to a high state of stress and anxiety and is also part of submissive behavior. Complex behaviors are excellent indicators of the state of anxiety of animals. NTT has been shown to be sensitive to pharmacological and genetic manipulation9, making it a valuable tool for studying the neural basis of anxiety and related disorders.
Zebrafish are a highly social species, so we can measure a wide range of social behaviors. The shoaling test (ST) and the social preference test (SPT) are the most used assays to assess social behavior10. The ST measures the tendency of fish to group together11 by quantifying their spatial behavior and movement patterns. ST is useful for studying group dynamics, leadership, social learning, and understanding the social behavior of many fish species12. The SPT in adult zebrafish was adapted from Crawley&#39;s preference for social novelty test for mice13&#160;and quickly became a popular behavioral assay for the study of social interaction in this model species14. These two tests have also been adapted for use in drug screening assays and have shown promise for identifying novel compounds that modulate social behavior15,16.
In general, behavioral assays in adult zebrafish are powerful tools that can provide valuable information on the behavior mechanisms or the neurophenotypes of active compounds and abused drugs17. This protocol details how to implement these behavioral tools7 with basic material resources and how to apply them in toxicity assays to characterize the effects of a wide range of neuroactive compounds. In addition, we will see that the same tests can be applied to assess the neurobehavioral effects of acute exposure to a neuroactive compound (methamphetamine) but also to characterize these effects after chronic exposure to environmental concentrations of a pesticide (glyphosate).

Författare i fokus: Bedömning av miljö- och läkemedelsinducerade beteendeförändringar hos vuxna zebrafiskar 

Neurotoxicitetsbedömning hos vuxna Danio rerio med hjälp av ett batteri av beteendetester i en enda tank

Our main research aims to identify the behavioral effects induced by environmental pollutants and drugs in fish. Also trying to understand the mechanisms leading to these effects. A social and anxiety-like behavior cannot be conveniently addressed using embryos.We conduct studies targeting these behaviors in the adult Zebrafish model. This protocol details how to implement behavioral tools with basic material resources to facilitate the reproducibility of tests and harmonize the results obtained within the scientific community. The assays presented in this protocol have proven to be highly reproducible and sensitive to genetic and pharmacological manipulations, making them variable and noninvasive tools fostering the neural and molecular mechanisms underlying behavior.Once a behavioral effort has been identified, the most challenging part is to determine the specific modes of action leading to the observative efforts. Starting by the molecular initiating event where predictions can provide some clues on the molecular initiating event, we are still far from obtaining accurate predictions. We are currently interested in the determining the phase validity of the chemical models of Parkinson's disease developed in embryonic and adult Zebrafish by using MPTP and rotenone with the same, the kinematic of the gait-like movement or during an explosive movement like the acoustic evoked escape response, will be evaluated.

Watch this Scientific Journal Video about Experimental Tank Configuration and Video Recording Setup for Behavioral Testing in Adult Zebrafish at JoVE.com

Experimentell akvariekonfiguration och videoinspelningsinställning för beteendetestning hos vuxna zebrafiskar

Börja med att montera de fyrkantiga experimenttankarna, kamerorna och datorerna. För individuella beteendeanalyser av zebrafiskar. Fyll tanken med sju liter 28 grader Celsius väl syresatt fiskvatten.Justera tankens position framför kameran för att undvika förvrängda bilder. Använd LED-bakgrundsbelysningen, kontrollera belysningsinställningen och slå på kameran. Öppna sedan kamerahanteraren för att kontrollera tillgängligheten för jiggy-kameran på datorn.Starta jiggy-kamerakontrollprogramvaran och öppna alternativet Kamera. Välj sedan Monokromt läge och justera bildstorleken. Öppna kameraegenskaper och ställ in pixelklockan på max och bildfrekvensen på 30 bildrutor per sekund under Kamera.Justera sedan exponeringen. Nu under Bild, ställ in förstärkningen till noll och justera svartnivåerna. På fliken Storlek justerar du fönstermåtten till det område som behöver graveras.När du är klar stänger du kameraegenskaperna. Skapa sedan en allmän mapp för experimentsessionen för att spara kamerainställningarna och videorna. Klicka sedan på Arkiv, Spara parametrar och till fil för att spara inställningarna.När du har konfigurerat alla kameror öppnar du spela in videosekvens och väljer Skapa för att spara som en ny videofil. Välj maximalt antal bildrutor och ange 10 800 i rutan för bildrutor. Välj sedan Beräkna bildfrekvens.Introducera fiskarna en efter en i botten av experimenttankarna för beteendetester och klicka på Spela in för att starta inspelningen. För stimtestet, för in stimmet i botten av experimenttanken. För testet av social preferens placerar du fisken en efter en i botten av de olika experimentbassängerna.När en chattruta med maximalt antal bildrutor visas på skärmen klickar du på Acceptera. Välj sedan Stäng för att avsluta inspelningen. Slutligen tar du bort de fiskar som registrerats för beteendetester från experimentakvariet och separerar dem från naiva fiskar.