Knowledge Bank

Identifying muscle specific genes and understanding their functions contribute to a better understanding of muscle biology. A novel transmembrane protein gene, TMEM182, has been found to be expressed more abundantly than other genes in muscle tissues through our chicken microarray data. To understand the role of TMEM182 in muscle development in vitro, quail TMEM182 gene with a hemagglutinin (HA) tag identifying exogenous protein was amplified by PCR and cloned into a pQCXIP expression vector to transfect quail muscle 7 (QM7) cells. Using puromycin selection, one QM7 cell line containing a control empty vector and two QM7 cell lines with target TMEM182-HA gene were created. The expression of TMEM182-HA in the cell lines were confirmed by Western blot analysis. Although the TMEM182-HA protein band was predicted to be 26 kDa, the bands in the analysis appeared to be bigger at 30-40 kDa with hazy bands, which could be due to predicted glycosylations in the N-terminal portion of TMEM182. In addition, during the myogenic differentiation, overexpression of TMEM182 resulted in a near absence of myotube formation with dramatically reduced levels of myosin heavy chain protein, a myogenic marker, compared to control cells. These data clearly indicate the anti-myogenic function of TMEM182. Since TMEM182 has been found to inhibit myogenesis, TMEM182 can serve as a new target gene for selecting superior lines of food animals for high muscle growth, and for pharmaceutical approaches in decreasing TMEM182 activity to increase muscle growth. Our findings may be utilized as a foundation to explore the mechanisms by which TMEM182 inhibits myogenesis, and to understand muscle biology across many species including humans since TMEM182 is found from fish to mammals.