Systematic evaluation of the adaptability of the non-radioactive SUnSET assay to measure cardiac protein synthesis

Heart is a dynamic organ that undergoes remodeling in response to both physiological and pathological stimuli. One of the fundamental cellular processes that facilitates changes in the size and shape of this muscular organ is the protein synthesis. Traditionally changes in cardiac protein synthesis...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2018-03, Vol.8 (1), p.4587-11, Article 4587
Main Authors: Ravi, Venkatraman, Jain, Aditi, Ahamed, Faiz, Fathma, Nowrin, Desingu, Perumal Arumugam, Sundaresan, Nagalingam R.
Format: Article
Language:English
Subjects:
13/1
13/106
13/109
13/51
14
14/19
631/1647/2196/2197
692/4019/592/1540
Adaptability
Cardiomyocytes
Cell lines
Dendrites
Heart
Heart diseases
Humanities and Social Sciences
Hypertrophy
Lymphocytes T
multidisciplinary
Neonates
Protein biosynthesis
Protein synthesis
Proteins
Radioactive tracers
Science
Science (multidisciplinary)
Western blotting
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Heart is a dynamic organ that undergoes remodeling in response to both physiological and pathological stimuli. One of the fundamental cellular processes that facilitates changes in the size and shape of this muscular organ is the protein synthesis. Traditionally changes in cardiac protein synthesis levels were measured by radiolabeled tracers. However, these methods are often cumbersome and suffer from radioactive risk. Recently a nonradioactive method for detecting protein synthesis under in vitro conditions called the Surface Sensing of Translation (SUnSET) was described in cell lines of mouse dendrites and T cells. In this work, we provide multiple lines of evidence that the SUnSET assay can be applied to reliably detect changes in protein synthesis both in isolated neonatal primary cardiomyocytes and heart. We successfully tracked the changes in protein synthesis by western blotting as well as immunohistochemical variants of the SUnSET assay. Applying the SUnSET assay, we measured the cardiac protein synthesis during the different ages of mice. Further, we successfully tracked the increase in cardiac protein synthesis during different stages of a well-established model for pathological hypertrophy. Overall, we propose SUnSET assay as a simple, reliable and robust method to measure protein synthesis in the cardiac milieu.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-22903-8