Experimental Protocols

The availability of standardized operating procedures (SOPs) to unify experimental protocols used to test the effects of new treatments in animal models is a step that will undoubtedly improve the comparability of studies from different laboratories.

One of the goals of TREAT-NMD is to create a collection of SOPs that can be used as guidelines in the conduct of preclinical studies on mice and dogs. In full collaboration with the Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center at Children's National Medical Center in Washington DC (www.wellstone-dc.org/dod/ProjectsandCores/Murine
drugscreeningandtransgeniccore/tabid/288/Default.aspx) and with the US National Institutes of Health (NIH)-Wellstone Muscular Dystrophy Cooperative Network (www.wellstonemdcenters.nih.gov), two meetings were hosted with specialists from all over the world to create a set of SOPs for a number of experimental protocols in the mdx mouse and GRMD dog models. These workshops were generously supported by Foundation to Eradicate Duchenne Inc (www.duchennemd.org), the US National Institutes of Health (www.wellstonemdcenters.nih.gov) and TREAT-NMD. These workshops were described in a meeting report published in Neuromuscular Disorders and available here.

These SOPs are not meant to be mandatory but are designed to be a point of reference and should not prevent innovation and further improvement of the existing protocols.

The SOPs that have already been completed can be downloaded from this site and others will be added in the future. To ensure reproducibility and quality, each SOP has been drawn up by a group of independent researchers (listed as authors in each protocol) and will be updated on a regular basis. 
 
If you wish to assist us by providing us with feedback about the content of these protocols, please leave your email address here and thick the SOPs of interest, and we will contact you for your feedback. Please be rest assured we will not pass your email address onto anyone else and it will only be used for the purpose we have specified.

• Behavioral and locomotor measurements using open field animal activity monitoring system
• Whole body tension measurements
• The recovery score to evaluate therapy efficacy in NMD: a common, quantitative and comparative scoring system
• Use of treadmill and wheel exercise for impact on mdx mice phenotype
• Use of grip strenght meter to assess the limb strenght of mdx mice
• Measuring isometric force of isolated mouse skeletal muscles in situ
• Respiratory system evaluation
• Use of patch-clamp technique to study voltage-independent cationic channels in normal and dystrophic skeletal muscle fibres
• Non-invasive echocardiographic assessment of cardiac function in the mdx mouse model
• Measuring isometric force decrement after eccentric contraction in vivo in GRMD dogs
• Measuring tibiotarsal joint angle to assess contractures in vivo in GRMD dogs
• Measuring tetanic isometric force at the tibiotarsal joint in vivo in GRMD dogs
• Left ventricular pressure-volume (P/V) loops
• Use of treadmill and wheel exercise to assess dystrophic state
• Measuring isometric force of isolated mouse muscles in vitro
• Workshop report in Neuromuscular Disorders - preclinical testing for DMD
• Quantitative determination of muscle fiber diameter (minimal Feret’s diameter) and percentage of centralized nuclei
• Determination of hydroxyproline content as a measure of fibrosis in nondystrophic and dystrophic skeletal muscle