title: Delivery of recombinant adeno-associated virus vectors to rat diaphragm muscle via direct intramuscular injection.
abstract: The diaphragm is the most important inspiratory muscle in all mammals, and ventilatory insufficiency caused by diaphragm dysfunction is the leading cause of morbidity and mortality in many genetic and acquired diseases affecting skeletal muscle. Currently, pharmacological inhibitors, genetically modified animals, and invasive procedures are used to study disorders affecting the diaphragm. However, these methodologies can be problematic because of off-target drug effects and the possible nonphysiological consequences of lifelong genetic alterations. Therefore, alternative methods to study this important respiratory muscle are needed. To resolve this, we have developed a methodology to deliver recombinant adeno-associated virus (rAAV) vectors to the rat diaphragm via direct intramuscular injection. We hypothesized that by direct injection of rAAV into the muscle we can selectively target the diaphragm and establish a novel experimental method for studying signaling pathways and also provide a strategy for effectively using rAAV to protect the diaphragm against disease. This report describes the methods and evidence to support the use of rAAV as a therapeutic intervention to study rat diaphragm biology during conditions that promote diaphragm dysfunction.
authors.affiliation: 1 Department of Applied Physiology and Kinesiology, Center for Exercise Science, University of Florida , Gainesville, FL 32611.
authors.forename: Ashley J
authors.initials: AJ
authors.lastname: Smuder
authors.forename: Darin J
authors.initials: DJ
authors.lastname: Falk
authors.forename: Kurt J
authors.initials: KJ
authors.lastname: Sollanek
authors.forename: W Bradley
authors.initials: WB
authors.lastname: Nelson
authors.forename: Scott K
authors.initials: SK
authors.lastname: Powers
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D003964:Diaphragm / Q000378:metabolism* / Q000502:physiology; D005260:Female; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000009:adverse effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007273:Injections, Intramuscular; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D049452:Green Fluorescent Proteins