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Rigorous Validation of Paw Preference Using Three Complementary Behavioral Assays in Sprague Dawley Rats.

Dipesh Pokharel1,2,3, Khoi Le3, Dilshan Beligala4

  • 1Department of Neurosciences and Psychiatry, The University of Toledo College of Medicine and Life Sciences, Toledo, OH, USA, 43614.

Biorxiv : the Preprint Server for Biology
|February 23, 2026
PubMed
Summary
This summary is machine-generated.

Three common paw preference tests in Sprague Dawley rats show reliable and consistent results. Paw preference is stable across age and sex, making these tests valuable for preclinical research on motor function.

Keywords:
Collins TestFunctional cerebral asymmetriesHandednessHemispheric dominanceLimb dominancePawedness Trait TestStaircase Test

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Area of Science:

  • Neuroscience and Animal Behavior
  • Rodent Models in Research
  • Behavioral Neuroscience

Background:

  • Paw preference, a measure of lateralization, is crucial for assessing motor function in rodents.
  • Understanding paw preference consistency is vital for reliable preclinical research.
  • Existing behavioral tests for paw preference require rigorous validation.

Purpose of the Study:

  • To evaluate the consistency and reliability of three common paw preference tests: Collins, Staircase, and Pawedness Trait Test.
  • To assess the influence of age and sex on paw preference in Sprague Dawley rats.
  • To establish validated behavioral assays for preclinical studies.

Main Methods:

  • Thirty Sprague Dawley rats underwent the Collins, Staircase, and Pawedness Trait Tests.
  • Paw usage was recorded, and laterality index was calculated for each test.
  • Statistical analyses included ANOVA and Cohen's Kappa to assess inter-test agreement and variations.

Main Results:

  • All three tests provided comparable measures of paw preference (ANOVA, p = 0.801).
  • Strong inter-test agreement was observed (Fleiss' kappa, κ = 0.761).
  • Paw preference showed no significant variation across different ages or sexes.

Conclusions:

  • The Collins, Staircase, and Pawedness Trait Tests are consistent and reliable for assessing paw preference in rats.
  • These validated tests can be utilized in preclinical research for motor asymmetry and neurodegenerative diseases.
  • The findings support the use of these assays for studying conditions like stroke and cerebral palsy.