Monitoring Fine and Associative Motor Learning in Mice Using the Erasmus Ladder

Summary

This article presents a protocol that allows a non-invasive and automated assessment of fine motor performance, as well as adaptive and associative motor learning upon challenges, using a device called the Erasmus Ladder. Task difficulty can be titrated to detect motor impairment ranging from major to subtle degrees.

Abstract

Behavior is shaped by actions, and actions necessitate motor skills such as strength, coordination, and learning. None of the behaviors essential for sustaining life would be possible without the ability to transition from one position to another. Unfortunately, motor skills can be compromised in a wide array of diseases. Therefore, investigating the mechanisms of motor functions at the cellular, molecular, and circuit levels, as well as understanding the symptoms, causes, and progression of motor disorders, is crucial for developing effective treatments. Mouse models are frequently employed for this purpose.

This article describes a protocol that allows the monitoring of various aspects of motor performance and learning in mice using an automated tool called the Erasmus Ladder. The assay involves two phases: an initial phase where mice are trained to navigate a horizontal ladder built of irregular rungs ("fine motor learning"), and a second phase where an obstacle is presented in the path of the moving animal. The perturbation can be unexpected ("challenged motor learning") or preceded by an auditory tone ("associative motor learning"). The task is easy to conduct and is fully supported by automated software.

This report shows how different readouts from the test, when analyzed with sensitive statistical methods, allow fine monitoring of mouse motor skills using a small cohort of mice. We propose that the method will be highly sensitive to evaluate motor adaptations driven by environmental modifications as well as early-stage subtle motor deficits in mutant mice with compromised motor functions.

Introduction

A variety of tests have been developed to assess motor phenotypes in mice. Each test gives information on a specific aspect of motor behavior¹. For example, the open field test informs on general locomotion and anxiety state; the rotarod and walking beam tests on coordination and balance; footprint analysis is about gait; the treadmill or running wheel on forced or voluntary physical exercise; and the complex wheel is about motor skill learning. To analyze mouse motor phenotypes, investigators must perform these tests sequentially, which involves a lot of time and effort and often several animal cohorts. If there is information at the cellular ....

Protocol

In the current study, adult (2-3 months old) C57BL/6J mice of both sexes were used. Animals were housed two to five per cage with ad libitum access to food and water in an animal unit under observation and maintained in a temperature-controlled environment on a 12 h dark/light cycle. All procedures were conducted in accordance with the European and Spanish regulations (2010/63/UE; RD 53/2013) and were approved by the Ethical Committee of the Generalitat Valenciana and the animal welfare committee of the Universidad Miguel Hernández.

1. Behavioral room setup

1. Reserve the behavioral testing room every....

Results

The Erasmus Ladder device, setup, and protocol applied are presented in Figure 1. The protocol consists of four undisturbed and four challenge sessions (42 trials each). Each trial is one run on the ladder between the starting and ending goal boxes. At the beginning of the session, a mouse is placed in one of the starting boxes. After a set time of 15 ± 5 s ("resting" state), the light is turned on (cue 1, for a maximum of 3 s). A light air cue (cue 2, 45 s maximum) is then appl.......

Discussion

The Erasmus Ladder presents major advantages for motor phenotype assessment beyond current approaches. Testing is easy to conduct, automated, reproducible, and allows researchers to assess various aspects of motor behavior separately using a single mouse cohort. In the current study, reproducibility allowed the generation of robust data with a small number of WT mice taking advantage of the features of the device, experimental design, and analysis methods. For instance, when compared to traditional beam-walk assays, the .......

Materials

Name	Company	Catalog Number	Comments
C57BL/6J mice (Mus musculus)	Charles Rivers
Erasmus Ladder device	Noldus, Wageningen, Netherlands
Erasmus Ladder 2.0 software	Noldus, Wageningen, Netherlands
Excel software	Microsoft
Sigmaplot software	Systat Software, Inc.