Anxiety Model In Adult Zebrafish

One of the central questions in biological psychiatry is how genes, molecular pathways, and patterns of connectivity in the brain produce and modulate anxiety behavior. The zebrafish (Danio rerio) has emerged as a useful new model for studying the behavioral and molecular mechanisms of brain disorders.

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ANXIETY MODEL IN ADULT ZEBRAFISH 2

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Introduction:

One of the central questions in biological psychiatry is how genes, molecular pathways, and patterns of connectivity in the brain produce and modulate anxiety behavior. 

Through the use of numerous behavioral paradigms, genetic/pharmacological screens, and neuroimaging, animals have been extensively used to model anxiety pathogenesis.

The zebrafish (Danio rerio) has emerged as a useful new model for studying the behavioral and molecular mechanisms of brain disorders.

Mode of Induction:

  • Modeling of stress and anxiety in adult zebrafish (Danio rerio) is increasingly utilized in neuroscience research and central nervous system (CNS) drug discovery.
  • Chronic unpredictable stress-like behavior induces anxiety-like behavior in young zebrafish.
  • Representing the most commonly used zebrafish anxiety models, the Predator avoidance (PA) focuses on zebrafish response to potentially threatening stimuli, whereas the light-dark test (LDT) is based on fish scototaxis (innate preference for dark vs. bright areas).

Analyzing Parameters for Anxiety Model:

1) Predator avoidance

The following endpoints are typically recorded:

  • The latency to enter the same arm as the predator (s),
  • Time spent in the same arm as the predator (s), 
  • Time spent in the arms without the predator (s) number of transitions (entries) into the same arm as the predator
  • Number of transitions (entries) into the arms without the predator.

2) Light-dark box

The following endpoints are typically recorded in this test: 

  • Latency to enter the white half (s),
  • Time spent in the white half (s), 
  • The number of entries to the white half of the apparatus.

3) Histopathology of Brain:

The following endpoints are typically recorded in this test: 

  • Swelling of neural tissues and cells.
  • The concentration of inflammatory cells in the brain.
  • Darkening of neural tissues and cells.

4) Tissue Collection, Protein Preparation, and Differential Proteomic Analysis:

  • Total brain proteins were extracted following homogenization, precipitation and solubilization.
  • Protein was quantified based on the Amido black assay method against BSA
  • Differential proteomic analyses between the control and test (CUS) samples were performed using 2-dimensional gel electrophoresis (2DE) and mass spectrometric analysis.

5) SDS-PAGE & Immunoblotting

6) Gene Expression Analysis by Real-time PCR

Conclusion:

This simple stress-induced anxiety and related affective disorder model can be used for the routine screening of compounds positive in cell-based assays for CNS drug discovery, as well as for in vivo validation of the lead molecules for their efficacy in various neurological and psychiatric disorders.
A better understanding of stress-induced alterations in the brain proteome profile might facilitate the identification of new molecular targets for CNS drug development.

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