All Phenotypes Uvrag<Tn(pb-Act-RFP)1.100107015-HRAZhu> MGI Mouse

increased myocardial fiber size ( J:219546 )

• by 10 months of age, the cross-sectional area of individual cardiomyocytes is significantly increased relative to wild-type controls

enlarged heart ( J:219546 )

• by 10 months of age, mutant hearts appear to be larger than wild-type

cardiac interstitial fibrosis ( J:219546 )

• by 10 months of age, mutant hearts show increased interstitial cardiac fibrosis relative to wild-type controls

abnormal cardiovascular system physiology ( J:219546 )

• at 10 months of age, mutant hearts accumulate autophagosomes and autophagic flux, based on changes in LC3 II protein levels elicited by chloroquine treatment, is impaired

decreased cardiac muscle contractility ( J:219546 )

• by 10 months of age, ejection fraction (EF) and percent fractional shortening (%FS) are reduced, suggesting impaired cardiac function

abnormal heart echocardiography feature ( J:219546 )

• by 10 months of age, the end-diastolic left ventricular dimension (LVEDD) and the end-systolic left ventricular dimension (LVESD) are significantly increased relative to wild-type controls

cardiomyopathy ( J:219546 )

• homozygotes are viable, fertile, and developmentally normal but develop age-related cardiomyopathy, evident by 10 months of age

• no alterations in cardiac structure are noted at 2 and 6 months of age

heart inflammation ( J:219546 )

• immunohistochemical staining of mutant hearts revealed that inflammatory cell markers CD45 (leucocytes) and CD45R (B cells) are significantly increased relative to wild-type hearts

• heart inflammation is already detectable at 6 months of age when cardiac structure and function are normal

cardiac interstitial fibrosis ( J:219546 )

• by 10 months of age, mutant hearts show increased interstitial cardiac fibrosis relative to wild-type controls

abnormal lysosome morphology ( J:219546 )

• at 10 months of age, mutant hearts display accumulation of lysosomal markers LAMP-1 and LAMP-2, suggesting that impaired autophagosome clearance is associated with increased lysosome numbers

• in contrast, cathepsin D, a lysosomal protease, is not significantly increased in mutant hearts

abnormal autophagy ( J:219546 )

• at 10 months of age, mutant hearts accumulate autophagosomes, as shown by enhanced protein abundance of LC3 II; LC3 immunofluorescence and ultrastructural analysis confirmed accumulation of autophagic vacuoles in mutant hearts

• p62 (a marker for autophagic flux), is significantly increased in mutant hearts, indicating impaired autophagic flux

• the observed increase of LC3 II protein levels in mutant hearts is not further enhanced by treatment with chloroquine (an inhibitor of autophagosome-lysosome fusion)

• mutant MEFs show significantly increased LC3 II protein levels; however, the increase in LC3 II protein levels is not further enhanced by bafilomycin A1 (an autophagosome-lysosome fusion inhibitor), suggesting that autophagic flux is impaired in mutant MEFs

• ultrastructural analysis of mutant MEFs revealed clustered double-membrane autophagosomes and complex vacuoles containing multiple layers of membrane

increased cardiomyocyte apoptosis ( J:219546 )

• by 10 months of age, mutant hearts show significantly increased apoptosis, as shown by TUNEL analysis