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Author Notes:

Correspondence: Grace K. Pavlath, PhD, Department of Pharmacology, Emory University, 1510 Clifton Road, Room 5027, Atlanta, GA 30322, USA. Tel.: +1 404 727 3353; fax: +1 404 727 0365; e‐mail: gpavlat@emory.edu

Anita H. Corbett, PhD, Department of Biology, Emory University, 1510 Clifton Road, Room 1021, Atlanta, GA 30322, USA. Tel.: +1 404 727 4546; fax: +1 404 7272880; e‐mail: accorbe2@emory.edu

AAC participated in conception and design, collection and assembly of data, data analysis and interpretation, financial support, manuscript writing, and final approval of manuscript.

ED and DD were involved in sample preparation, data analysis and interpretation of proteomic experiments, and final approval of manuscript.

NTS carried out data analysis and interpretation of proteomic experiments, manuscript writing, and final approval of manuscript.

AHC took part in conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript.

GP participated in conception and design, data analysis and interpretation, financial support, manuscript writing, and final approval of manuscript.

We thank the Emory University School of Medicine Core Facility for Flow Cytometry and the Emory Integrated Proteomics Core for experimental assistance.

Conflict of interest: none declared.

Subjects:

Research Funding:

This work was supported by grants to GKP (AR062483), to AAC (AR067645), and a training grant (T32GM008367) from the National Institutes of Health.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Cell Biology
  • Geriatrics & Gerontology
  • aging
  • brain
  • myonuclei
  • nuclear isolation
  • pro-teome
  • skeletal muscle
  • AGED SKELETAL-MUSCLE
  • GENE-EXPRESSION
  • PEPTIDE IDENTIFICATION
  • REVEALS
  • NUCLEI
  • CELLS
  • PHOSPHORYLATION
  • QUANTIFICATION
  • FRACTIONATION
  • PURIFICATION

SBiochemical isolation of myonuclei employed to define changes to the myonuclear proteome that occur with aging

Tools:

Journal Title:

Aging Cell

Volume:

Volume 16, Number 4

Publisher:

, Pages 738-749

Type of Work:

Article | Final Publisher PDF

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.

Copyright information:

© 2017 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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