Protocol Citation: Tim Sampson, Ian Krout 2025. Hindlimb Scoring. protocols.io https://dx.doi.org/10.17504/protocols.io.eq2ly4n2mlx9/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 31, 2025
Last Modified: August 01, 2025
Protocol Integer ID: 223847
Keywords: ASAPCRN, Behavior, Mice, PD, Parkinsons, Hindlimb, hindlimb clasping of mice, dopamine loss in the mouse model1, dopamine loss, hindlimb, hindlimb clasping, baseline reading of hindlimb, mice, mouse, pd like progression, mouse model1, neurobehavior, measure for pd, pd, striatal dysfunction, clasping assessment, assessment in mice, pd
Funders Acknowledgements:
Aligning Science Across Parkinson’s
Grant ID: ASAP-020495
Aligning Science Across Parkinson’s
Grant ID: ASAP-000375
Heritage Medical Research Institute
Abstract
This protocols detail hindlimb clasping assessment in mice as a measure for PD-like pathology and neurobehavior. Hindlimb clasping has been used in many studies as a measure of PD like progression as the extent of clasping correlates with dopamine loss in the mouse model1. The below method is an adapted protocol from Lieu 2013.
Materials
1. Standard mouse cage housing mice for assessment.
2. Two independent scorers.
Troubleshooting
Acclimation
Habituate animals to behavior testing room for 1 hour prior to start of assay.
Set-up
Find 2 independent scorers who are educated on the scoring system as detailed below.
Assessment
Start with the first cage and begin by lifting one mouse at a time by the tail, positioning the
mouse so that those scorers are viewing the stomach – mice should be held
~1.5” from the base of the tail and ~2’ over the wire top of their home cage
(away from the food basin).
Suspend the mice from ~5 seconds while those scoring observe and make their assessment.
Move on the next mouse in the cage until all mice have been scored in a cage, move onto next cage.
Repeat until all mice have been scored.
Scoring
Scoring is done for each mouse on a scale from 0 – 3.
A 0 is considered a pre-symptomatic score and is observed when the mouse is suspended by the tail and the hindlimb presents a normal spay, i.e., the animals freely move both limbs – extending them outwards (Figure 1).
Figure 1. A score of 0. Full splaying is observed, free movement of hindlimbs.
A 1 is observed when the mouse is suspended by the tail and one or both limbs show partial inward clasping towards the midline for ~50% of the observation time, or the limb of one side shows full clasping towards the midline for <50% of the observation time. A 1 should also be scored if trembling of the limbs is observed during the tail suspension (Figure 2).
Figure 2. A score of 1. Partial clasping either on one or both sides or full clasping on
one side.
A 2 is observed when the mouse is suspended by the tail and both limbs show partial clasping towards the midline for >75% of the observation time, or one side shows full clasping towards the midline for the duration of the observation time. In both cases, some flexibility of the hindlimbs is observed, but most of the time clasping of hindlimbs (one or both) is seen (Figure 3).
Figure 3. A score of 2. Partial clasping either on one or both sides or full clasping on
one side, duration determines a difference between 1-2 as well as the amount of
flexibility.
A 3 is considered as severe pathology and is observed when the mouse is suspended by the tail and both limbs immediately clasp and remain clasped for the duration of observation. In this case, there is no flexibility observed for the hindlimbs (Figure 4).
Figure 4. A score of 3. Full clasping of both hindlimbs is observed.
Protocol references
1. Lieu, Christopher & Chinta, Shankar & Rane, Anand & Andersen, Julie.
(2013). Age-Related Behavioral Phenotype of an Astrocytic Monoamine Oxidase-B
Transgenic Mouse Model of Parkinson’s Disease. PloS one. 8. e54200.
10.1371/journal.pone.0054200.