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Related Experiment Video

Updated: Aug 20, 2025

Light/dark Transition Test for Mice
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Exploring the light/dark box test: Protocols and implications for neuroscience research.

Rodrigo Campos-Cardoso1, Lívea Dornela Godoy2, Willian Lazarini-Lopes1

  • 1Departamento de Neurociências e Ciências do Comportamento, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Avenida Bandeirantes, 3900, Cidade Universitária, 14040-901 Ribeirão Preto, SP, Brazil; Instituto de Neurociências e Comportamento (INeC), Avenida Bandeirantes, 3900, Ribeirão Preto, SP 14049-901, Brazil.

Journal of Neuroscience Methods
|November 21, 2022
PubMed
Summary

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Light intensity in the light-dark box test significantly impacts rodent anxiety-like behavior. Researchers recommend a neutral 65 lux condition for accurate drug and stressor effect evaluation in psychiatric disorder research.

Area of Science:

  • Neuroscience
  • Behavioral Science
  • Psychopharmacology

Background:

  • Advances in understanding psychiatric disorders rely on animal models and neurobiological research.
  • Rodent anxiety-like behaviors are studied using neuroethological tests like the light-dark box (LDB).
  • LDB test protocols can influence behavioral outcomes and hypothalamic-pituitary-adrenal (HPA) axis activation.

Purpose of the Study:

  • To investigate how different light intensities and prior stress influence rodent behavior in the LDB test.
  • To determine optimal conditions for evaluating anxiolytic/anxiogenic effects in the LDB.
  • To understand factors affecting exploratory and risk assessment behaviors in rodents.

Main Methods:

  • Adult male Wistar rats were exposed to the LDB under varying illumination (25/0, 65/0, 330/0 lux).
Keywords:
AnxietyHPA axisLight intensityLight-dark boxNeuroethologyRisk assessmentStress

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  • Initial compartment positioning and prior stress (Elevated Plus Maze or restraint stress) were varied.
  • Behavior was recorded over 15 minutes, analyzed in 5-minute blocks.

    • Main Results:

    • Exploration of the lit compartment decreased with higher light intensity.
    • Prior stress or dark compartment entry reduced lit compartment exploration.
    • Low light intensity increased exploration; high intensity decreased it. No significant changes in serum corticosterone were observed.

    Conclusions:

    • Light intensity and test duration critically affect LDB results, potentially confounding anxiolytic/anxiogenic assessments.
    • A 65/0 lux condition appears neutral for evaluating drug effects or prior aversive stimuli.
    • Understanding these behavioral factors is vital for advancing psychiatric disorder research and treatment.