Limits...
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.

Show MeSH

Related in: MedlinePlus

Knockdown of the ∼70-kDa immunoreactive species of Panx3 inhibits skeletal muscle myoblast proliferation without inducing their differentiation. Transfection of two Panx3 shRNAs (sh63 and sh64) resulted in a significant reduction of the ∼70-kDa immunoreactive species of Panx3 compared with the control scramble shRNA (Ctl) (A and B) without modifying the levels of Panx1 nor the ∼43-kDa species of Panx3 (A). BrdU incorporation assay showed that knockdown of the ∼70-kDa species reduces HSMM proliferation in growth medium (GM) (C) without triggering their differentiation (D). Undifferentiated (Day 0) and differentiated (Day 6 in DM) HSMM were used as comparison. Tubulin was used as a loading control. *, p < 0.05 compared with the Ctl shRNA; #, p < 0.001 compared with Day 0.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4215249&req=5

Figure 9: Knockdown of the ∼70-kDa immunoreactive species of Panx3 inhibits skeletal muscle myoblast proliferation without inducing their differentiation. Transfection of two Panx3 shRNAs (sh63 and sh64) resulted in a significant reduction of the ∼70-kDa immunoreactive species of Panx3 compared with the control scramble shRNA (Ctl) (A and B) without modifying the levels of Panx1 nor the ∼43-kDa species of Panx3 (A). BrdU incorporation assay showed that knockdown of the ∼70-kDa species reduces HSMM proliferation in growth medium (GM) (C) without triggering their differentiation (D). Undifferentiated (Day 0) and differentiated (Day 6 in DM) HSMM were used as comparison. Tubulin was used as a loading control. *, p < 0.05 compared with the Ctl shRNA; #, p < 0.001 compared with Day 0.

Mentions: The ∼70-kDa Panx3 immunoreactive band detected in undifferentiated HSMM and skeletal muscle tissue has also been detected in murine organs (3), murine skin (6), rat organotypic epidermis (6, 7), and rat male reproductive tract (8). In organotypic epidermis, this band was recognized by three different anti-Panx3 antibodies and also corresponds to a glycoprotein (7). We now demonstrate that the endogenous levels of the ∼70-kDa immunoreactive species of Panx3 can be significantly knocked down using two different shRNAs against Panx3 (Fig. 9, A and B), which provides further evidence that it corresponds to a Panx3 species. Panx1 levels and Panx3 lower Mr species remained very low or below detectable levels in cells transfected with these Panx3 shRNAs (Fig. 9A).


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)

Knockdown of the ∼70-kDa immunoreactive species of Panx3 inhibits skeletal muscle myoblast proliferation without inducing their differentiation. Transfection of two Panx3 shRNAs (sh63 and sh64) resulted in a significant reduction of the ∼70-kDa immunoreactive species of Panx3 compared with the control scramble shRNA (Ctl) (A and B) without modifying the levels of Panx1 nor the ∼43-kDa species of Panx3 (A). BrdU incorporation assay showed that knockdown of the ∼70-kDa species reduces HSMM proliferation in growth medium (GM) (C) without triggering their differentiation (D). Undifferentiated (Day 0) and differentiated (Day 6 in DM) HSMM were used as comparison. Tubulin was used as a loading control. *, p < 0.05 compared with the Ctl shRNA; #, p < 0.001 compared with Day 0.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: Knockdown of the ∼70-kDa immunoreactive species of Panx3 inhibits skeletal muscle myoblast proliferation without inducing their differentiation. Transfection of two Panx3 shRNAs (sh63 and sh64) resulted in a significant reduction of the ∼70-kDa immunoreactive species of Panx3 compared with the control scramble shRNA (Ctl) (A and B) without modifying the levels of Panx1 nor the ∼43-kDa species of Panx3 (A). BrdU incorporation assay showed that knockdown of the ∼70-kDa species reduces HSMM proliferation in growth medium (GM) (C) without triggering their differentiation (D). Undifferentiated (Day 0) and differentiated (Day 6 in DM) HSMM were used as comparison. Tubulin was used as a loading control. *, p < 0.05 compared with the Ctl shRNA; #, p < 0.001 compared with Day 0.
Mentions: The ∼70-kDa Panx3 immunoreactive band detected in undifferentiated HSMM and skeletal muscle tissue has also been detected in murine organs (3), murine skin (6), rat organotypic epidermis (6, 7), and rat male reproductive tract (8). In organotypic epidermis, this band was recognized by three different anti-Panx3 antibodies and also corresponds to a glycoprotein (7). We now demonstrate that the endogenous levels of the ∼70-kDa immunoreactive species of Panx3 can be significantly knocked down using two different shRNAs against Panx3 (Fig. 9, A and B), which provides further evidence that it corresponds to a Panx3 species. Panx1 levels and Panx3 lower Mr species remained very low or below detectable levels in cells transfected with these Panx3 shRNAs (Fig. 9A).

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