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Standard Operating Procedures (SOP's) for Duchenne Animal Models

Standard Operating Procedures (SOP's) for Duchenne Animal Mo

Cardiac Protocols for Duchenne Animal Models

A workshop was organized by the National Heart, Lung, and Blood Institute (NHLBI), and sponsored by PPMD, in July 2014, in order to address Contemporary Cardiac Issues in Duchenne Muscular Dystrophy. This workshop resulted in 3 small groups; one of the groups was tasked to update and develop standard operating procedures (SOP’s) incorporating state of the art cardiac assessment and monitoring. Animal Model Small Group members included Dongsheng Duan (co-leader), Jill Rafael-Fortney (co-leader), Alison Blain, David Kass, Elizabeth McNally, Joseph Metzger, and Chris Spurney.

The standardization of operating procedures for cardiac research, and research involving cardiac assessment and monitoring in Duchenne animal models, hopes to improve the comparability of research studies performed in multiple laboratories. These SOP’s are not meant to be mandatory, but used as a point of reference amongst laboratories. They are meant to be "living documents," incorporating innovation and further improvement as they are implemented. Concerns/questions/suggestions should be directed to Kathi Kinnett at kathi@parentprojectmd.org.

All SOP's were created/updated June 2015, unless otherwise indicated.

1. General protocols
1.1. Collection and preservation of heart tissue for histological and biochemical studies
1.2. Immunofluorescence staining on unfixed cryosections
1.3. Monoclonal dystrophin antibodies for heart tissue immunostaining and western blot

2. Functional assay protocols related to the rodent model
2.1. Mouse heart Evans blue dye (EBD) uptake assay
2.2. Mouse dobutamine stress survival assay
2.3. Cardiovascular treadmill endurance assay in conscious mice
2.4. Force assessment in multicellular cardiac preparations in mice
2.5. Rodent electrocardiogram (ECG) recording
2.6. Twelve-lead non-invasive mouse electrocardiography (ECG)
2.7. Echocardiography in Rodents
2.8. In vivo pressure-volume loop studies in mice
2.9. Closed-chest left ventricular (LV) hemodynamic assay with the Millar catheter
2.10. Closed-chest right ventricular (RV) hemodynamic assay with the Millar catheter
2.11. Magnetic resonance imaging (MRI) in rodents
2.12. Manganese Enhanced MRI (MEMRI)

3. Functional assay protocols related to the large animal model
3.1. Large mammal (canine) ECG
3.2. ECG in conscious dogs (Coming soon)

4. Rodent cardiac function reference values
4.1. Mouse ECG reference values (Coming soon)
4.2. Mouse left ventricle PV loop reference values (Coming soon)

5. Large animal cardiac function reference values
5.1. Dog ECG reference values (Coming soon)
5.2. Dog echocardiography reference values (Coming soon)

Non-Cardiac SOP’s for Duchenne Animal Models

Non-cardiac SOP’s for Duchenne animal models are available on the TREAT-NMD website. Two meetings were hosted with specialists from all over the world to discuss and create this collection of SOPs, in full collaboration with the Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center at Children's National Medical Center in Washington DC and with the US National Institutes of Health (NIH)-Wellstone Muscular Dystrophy Cooperative Network, and with the generous support of the Foundation to Eradicate Duchenne Inc, the US National Institutes of Health and TREAT-NMD. These workshops were described in a meeting report published in Neuromuscular Disorders and available on the TREAT-NMD website.

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