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Pannexin 1 and pannexin 3 channels regulate skeletal muscle myoblast proliferation and differentiation.

Langlois S, Xiang X, Young K, Cowan BJ, Penuela S, Cowan KN - J. Biol. Chem. (2014)

Bottom Line: Using HSMM, we found that Panx1 expression promotes this process, whereas it was impaired in the presence of probenecid or carbenoxolone.Reduction of its endogenous expression using two Panx3 shRNAs significantly inhibited HSMM proliferation without triggering their differentiation.In summary, our results demonstrate that Panx1 and Panx3 are co-expressed in human skeletal muscle myoblasts and play a pivotal role in dictating the proliferation and differentiation status of these cells.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Surgery, Division of Paediatric Surgery, University of Ottawa, Children's Hospital of Eastern Ontario, Ottawa, Ontario K1H 8L1, Canada, Apoptosis Research Center, Children's Hospital of Eastern Ontario, Ottawa, Ontario K1H 8L1, Canada.

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Overexpression of Panx1 accelerates human primary skeletal muscle myoblast differentiation. HSMM were transfected with Panx1 (labeled in green) or GFP and placed in differentiation medium and examined 2 days later for MHC (labeled in red) expression and myotube formation. Representative pictures of three independent experiments are shown in A. When compared with cells expressing GFP, a higher proportion of Panx1-transfected cells were MHC-positive (B), and among those cells, a higher percentage contained two or more nuclei indicative of fusion (A, arrow), which was quantified in C. Blue = nuclei; bars = 50 μm. *, p < 0.05 compared with GFP.
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Figure 5: Overexpression of Panx1 accelerates human primary skeletal muscle myoblast differentiation. HSMM were transfected with Panx1 (labeled in green) or GFP and placed in differentiation medium and examined 2 days later for MHC (labeled in red) expression and myotube formation. Representative pictures of three independent experiments are shown in A. When compared with cells expressing GFP, a higher proportion of Panx1-transfected cells were MHC-positive (B), and among those cells, a higher percentage contained two or more nuclei indicative of fusion (A, arrow), which was quantified in C. Blue = nuclei; bars = 50 μm. *, p < 0.05 compared with GFP.

Mentions: Because Panx1 levels were increased during skeletal muscle differentiation, it is tempting to speculate that its expression promotes this process. To verify this hypothesis, Panx1 was overexpressed in HSMM and tested for its effect on their differentiation. HSMM were chosen for this assay as their ability to differentiate was found more consistent than that of SkMC. HSMM were thus transfected with either GFP or Panx1 and induced to differentiate for 2 days. Transfected cells were then examined for their expression of MHC and the formation of multinucleated myotubes as markers for terminal differentiation (Fig. 5A). After 2 days in differentiation media, a significant higher proportion of Panx1-transfected HSMM were MHC-positive (Fig. 5B) and contained two or more nuclei (Fig. 5C) when compared with control cells transfected with GFP, which did not show evidence of fusion yet. These results thus indicate that Panx1 promotes earlier skeletal muscle differentiation.


Pannexin 1 and pannexin 3 channels regulate skeletal muscle myoblast proliferation and differentiation.

Langlois S, Xiang X, Young K, Cowan BJ, Penuela S, Cowan KN - J. Biol. Chem. (2014)

Overexpression of Panx1 accelerates human primary skeletal muscle myoblast differentiation. HSMM were transfected with Panx1 (labeled in green) or GFP and placed in differentiation medium and examined 2 days later for MHC (labeled in red) expression and myotube formation. Representative pictures of three independent experiments are shown in A. When compared with cells expressing GFP, a higher proportion of Panx1-transfected cells were MHC-positive (B), and among those cells, a higher percentage contained two or more nuclei indicative of fusion (A, arrow), which was quantified in C. Blue = nuclei; bars = 50 μm. *, p < 0.05 compared with GFP.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215249&req=5

Figure 5: Overexpression of Panx1 accelerates human primary skeletal muscle myoblast differentiation. HSMM were transfected with Panx1 (labeled in green) or GFP and placed in differentiation medium and examined 2 days later for MHC (labeled in red) expression and myotube formation. Representative pictures of three independent experiments are shown in A. When compared with cells expressing GFP, a higher proportion of Panx1-transfected cells were MHC-positive (B), and among those cells, a higher percentage contained two or more nuclei indicative of fusion (A, arrow), which was quantified in C. Blue = nuclei; bars = 50 μm. *, p < 0.05 compared with GFP.
Mentions: Because Panx1 levels were increased during skeletal muscle differentiation, it is tempting to speculate that its expression promotes this process. To verify this hypothesis, Panx1 was overexpressed in HSMM and tested for its effect on their differentiation. HSMM were chosen for this assay as their ability to differentiate was found more consistent than that of SkMC. HSMM were thus transfected with either GFP or Panx1 and induced to differentiate for 2 days. Transfected cells were then examined for their expression of MHC and the formation of multinucleated myotubes as markers for terminal differentiation (Fig. 5A). After 2 days in differentiation media, a significant higher proportion of Panx1-transfected HSMM were MHC-positive (Fig. 5B) and contained two or more nuclei (Fig. 5C) when compared with control cells transfected with GFP, which did not show evidence of fusion yet. These results thus indicate that Panx1 promotes earlier skeletal muscle differentiation.

Bottom Line: Using HSMM, we found that Panx1 expression promotes this process, whereas it was impaired in the presence of probenecid or carbenoxolone.Reduction of its endogenous expression using two Panx3 shRNAs significantly inhibited HSMM proliferation without triggering their differentiation.In summary, our results demonstrate that Panx1 and Panx3 are co-expressed in human skeletal muscle myoblasts and play a pivotal role in dictating the proliferation and differentiation status of these cells.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Surgery, Division of Paediatric Surgery, University of Ottawa, Children's Hospital of Eastern Ontario, Ottawa, Ontario K1H 8L1, Canada, Apoptosis Research Center, Children's Hospital of Eastern Ontario, Ottawa, Ontario K1H 8L1, Canada.

Show MeSH
Related in: MedlinePlus