Biochemical isolation of myonuclei employed to define changes to the myonuclear proteome that occur with aging |
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| Authors: | Alicia A Cutler Eric B Dammer Duc M Doung Nicholas T Seyfried Anita H Corbett Grace K Pavlath |
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| Affiliation: | 1. Department of Pharmacology, Emory University, Atlanta, GA, USA;2. Graduate Program in Biochemistry, Cell and Developmental Biology, Emory University, Atlanta, GA, USA;3. Department of Biochemistry, Emory University, Atlanta, GA, USA;4. Department of Biology, Emory University, Atlanta, GA, USA |
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| Abstract: | Skeletal muscle aging is accompanied by loss of muscle mass and strength. Examining changes in myonuclear proteins with age would provide insight into molecular processes which regulate these profound changes in muscle physiology. However, muscle tissue is highly adapted for contraction and thus comprised largely of contractile proteins making the nuclear proteins difficult to identify from whole muscle samples. By developing a method to purify myonuclei from whole skeletal muscle, we were able to collect myonuclei for analysis by flow cytometry, biochemistry, and mass spectrometry. Nuclear purification dramatically increased the number and intensity of nuclear proteins detected by mass spectrometry compared to whole tissue. We exploited this increased proteomic depth to investigate age‐related changes to the myonuclear proteome. Nuclear levels of 54 of 779 identified proteins (7%) changed significantly with age; these proteins were primarily involved in chromatin maintenance and RNA processing. To determine whether the changes we detected were specific to myonuclei or were common to nuclei of excitatory tissues, we compared aging in myonuclei to aging in brain nuclei. Although several of the same processes were affected by aging in both brain and muscle nuclei, the specific proteins involved in these alterations differed between the two tissues. Isolating myonuclei allowed a deeper view into the myonuclear proteome than previously possible facilitating identification of novel age‐related changes in skeletal muscle. Our technique will enable future studies into a heretofore underrepresented compartment of skeletal muscle. |
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| Keywords: | aging brain myonuclei nuclear isolation proteome skeletal muscle |
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