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Identification and Characterization of TNF Receptor Associated Factor 3 Interacting Protein 3 (Traf3ip3) in Skeletal Muscle
Author(s) -
Golliher Ciara,
DaSilva Anjelica,
Waddell David
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.700.17
Subject(s) - myogenesis , denervation , skeletal muscle , biology , myocyte , microbiology and biotechnology , muscle atrophy , itga7 , reporter gene , gene expression , differential display , gene , endocrinology , genetics
Skeletal muscle atrophy is a physiological condition that results in decreased muscle size and strength and occurs in response to immobilization, denervation, spinal cord injury and aging. To better characterize the molecular genetic events of neurogenic atrophy, a previous study isolated gastrocnemius muscle from mice following 3 days and 14 days of sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue in order to identify novel neurogenic atrophy‐induced genes. The microarray data revealed that TNF Receptor Associated Factor 3 Interacting Protein 3 (Traf3ip3) is expressed in skeletal muscle and is induced in response to denervation. To confirm that Traf3ip3 is expressed in muscle cells, we cloned the Traf3ip3 cDNA from cultured muscle cells and performed quantitative PCR (qPCR) to assess expression levels in both proliferating and differentiated muscle cells. The qPCR results demonstrated that Traf3ip3 expression levels are relatively low in proliferating myoblasts but show dramatically elevated expression in differentiated myotubes. To characterize the transcriptional regulation of Traf3ip3, fragments of the proximal promoter located immediately upstream of the start of transcription were cloned and fused to a reporter gene. The reporter plasmids were then transfected into C 2 C 12 mouse muscle cells in combination with myogenic regulatory factor (MRF) expression plasmids, which resulted in differential effects on reporter gene activity. The discovery that Traf3ip3 is expressed in muscle combined with the observation that this gene is induced in response to neurogenic atrophy helps further our understanding of the molecular genetic events of skeletal muscle wasting. Support or Funding Information The work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .