Experimental protocols

In the field of neuromuscular diseases, the last decade witnessed an encouraging progress in the quest for molecules and treatments that help patient to increase their quality of life and raise hopes for a definitive cure.

With the availability of appropriate animal models and the increasing interest in research for rare diseases it is important to keep the quality of publications high, to implement the reproducibility and comparability of results across labs and to increase translatability to human clinical trials.

The following protocols were elaborated by groups of independent reseachers (listed in each proocol) and can be now downloaded for free from these pages. This effort was made possible by TREAT-NMD, Cure CMD, AFM, FED and by the unvaluable collaboration of all researchers involved.

Disease model 

Title

Last review date

General protocols

allThe recovery score: a common, quantitative and comparative scoring system2008
SMA miceInfluence of housing, feeding, and handling conditions on SMA mouse performance2015
DyWRecommendations on the care and handling of the MDC1A mouse model dyW2014

Histology

mdxQuantitative determination of muscle fibre diameter (minimal Feret’s diameter) and percentage of centralized nuclei (mdx)2014
DyWQuantitative determination of minimal Feret’s diameter, including the evaluation of the percentage of centralized nuclei, fiber numbers, cross-sectional area and the percentage of fibrosis (sampling and data for DyW mice)2014
mdxDetermination of hydroxyproline content as a measure of fibrosis2014
DyWHistopathology in Masson Trichrome stained muscle sections2014
mdxQuantification of histopathology in Haemotoxylin and Eosin stained muscle sections (sampling and data for mdx mice)2014
DyWHistopathology in Hematoxylin & Eosin stained muscle sections (sampling and data for DyW mice)2014
SMACell Body-Motor Neuron counts2017
SMAImaging neuromuscular junction pathology in mouse models of SMA2015
DyWNeuromuscular junction imaging on whole muscles and muscle sections in CMD animal models2016
DyWAssessment of peripheral nerve pathology in laminin-alpha-2-deficient mice2016
DyWInflammation quantification in the LAMA2dy-W mouse2014

Biochemistry-Molecular Biology

SMAELISA to quantify SMN protein levels2016
SMASemiquantitative Western Blot2016
SMAHuman and murine SMN transcript analysis by absolute real time PCR2016
SMAGenotyping protocol for commonly used SMA model mice2011
MDSerum Creatine Kinase analysis in mouse models of muscular dystrophy2017

Physiology

mdxMeasuring isometric force of isolated mouse muscles in vitro2015
mdxMeasuring isometric force of isolated mouse skeletal muscles in situ2016
mdxUse of grip strength meter to assess the limb strength of mdx mice2014
SMAGrip strength (SMA mice)2017
DyWUse of Grip Strength Meter to Assess the Limb Strength of dyw Mice2017
SMAMuscle function evaluation through isometric force measurements in mouse models for muscular atrophy2016
CMDNon-invasive measurement of ankle flexion/extension torque for longitudinal follow-up of strength in small rodents2017
DyWSingle muscle contractile measurements in vivo and in situ2015
mdxWhole body tension measurements2010
mdxRespiratory system evaluation2016
XLMTMRespiratory assessment of a canine model of XLMTM using non-invasive respiratory impedence plethysmography2017
mdxNon-invasive echocardiographic assessment of cardiac function in the mdx mouse model2015
mdxLeft ventricular pressure-volume (PV) loops in mdx2013
GRMDMeasuring tetanic isometric force at the tibiotarsal joint in vivo in GRMD dogs2015
GRMDMeasuring isometric force decrement after eccentric contraction in vivo in GRMD dogs2015
GRMDMeasuring tibiotarsal joint angle to assess contractures in vivo in GRMD dogs2015
GRMDEvaluation of Muscles by Magnetic Resonance Imaging (MRI)2015
XLMTMGait assessment in a canine model of XLMTM using an instrumented carpet2017

Functional tests

mdxBehavioural and Locomotor Measurements Using Open Field Animal Activity Monitoring System
see video on JoVe
2017
mdxUse of treadmill and wheel exercise for impact on mdx mice phenotype2014
mdxUse of treadmill and wheel exercise to assess dystrophic state2015
mdxThe use of hanging wire tests to monitor muscle strength and condition over time
see video on JoVe
2016
mdxThe use of four limb hanging tests to monitor muscle strength and condition over time
see video on JoVe
2016
SMAUse of pen test (balance beam) to assess motor balance and coordination in mice2016
SMABehavioral Phenotyping for Neonates: Hind Limb Suspension Test (a.k.a. Tube Test)2011
MDBehavioral Phenotyping for neonates: Righting reflex2014

Electrophysiology

SMASynaptic transmission alterations in SMA mice/Electrophysiology2016
SMACompound Muscle Activated Potential (CMAP) recordings from the triceps surae2016
mdxUse of patch-clamp technique to study voltage-independent cationic channels in normal and dystrophic skeletal muscle fibers2016