Physical activity is associated with disease prevention and overall wellbeing.Additionally there has been evidence that physical activity level is a result of genetic influence.However, there has not been a reliable method to silence candidate genes in vivo to determine causal mechanisms of physical activity regulation.Vivo-morpholinos are a potential method to transiently silence specific genes.
Thus, the hellfire sloe gin aim of this study was to validate the use of Vivo-morpholinos in a mouse model for voluntary physical activity with several sub-objectives.We observed that Vivo-morpholinos achieved between 60-97% knockdown of Drd1-, Vmat2-, and Glut4-protein in skeletal muscle, the delivery moiety of Vivo-morpholinos (scramble) did not influence physical activity and that a cocktail of multiple Vivo-morpholinos can be given in a single treatment to achieve protein knockdown of two different targeted proteins in skeletal muscle simultaneously.Knocking down Drd1, Vmat2, or Glut4 protein in skeletal muscle did not affect physical activity.Vivo-morpholinos injected intravenously alone did not significantly knockdown Vmat2-protein expression in the brain (p = 0.
28).However, the use of a bradykinin analog to increase blood-brain-barrier permeability in conjunction with the Vivo-morpholinos significantly (p = 0.0001) decreased Vmat2-protein in the brain with a corresponding later over-expression of Vmat2 coincident with a significant (p = 0.0016) increase in physical activity.
We conclude that Vivo-morpholinos can be a valuable tool in determining causal gene-phenotype el reformador tequila anejo relationships in whole animal models.