P2 nucleotide receptors on C2C12 satellite cells

Abstract

In developing muscle cells environmental stimuli transmitted by purines binding to the specific receptors are crucial proliferation regulators. C2C12 myoblasts express numerous purinergic receptors representing both main classes: P2X and P2Y. Among P2Y receptors we have found the expression of P2Y 1 , P2Y 2 , P2Y 4 , P2Y 6 and P2Y 12 family members while among P2X receptors P2X 4 , P2X 5 and P2X 7 were discovered. We have been able to show that activation of those receptors is responsible for ERK class kinase activity, responsible for regulation of cell proliferation pathway. We have also demonstrated that this activity is calcium dependent suggesting Ca 2+ ions as secondary messenger between receptor and kinase regulatory system. More specifically, we do suspect that in C2C12 myoblasts calcium channels of P2X receptors, particularly P2X 5 play the main role in proliferation regulation. In further development of myoblasts into myotubes, when proliferation is gradually inhibited, the pattern of P2 receptors is changed. This phenomenon is followed by diminishing of the P2Y 2 -dependent Ca 2+ signaling, while the mRNA expression of P2Y 2 receptor reminds still on the high level. Moreover, P2X 2 receptor mRNA, absent in myoblasts appears in myotubes. These data show that differentiation of C2C12 cell line satellite myoblasts is accompanied by changes in P2 receptors expression pattern.Abbreviations: 2MeSADP -2-methylthio-ADP; BzATP -3 0 -0(4-benzoyl)benzoyl ATP; [Ca 2+ ]i -intracellular Ca 2+ concentration; DMEM -Dulbecco's modified essential medium; ERK -Ras/extracellular signal-regulated kinase; FCS -fetal calf serum; GAPDH -glycerol 3-phosphate dehydrogenase; HS -horse serum; InsP 3 -inositol 1,4,5-trisphosphate; PLCphospholipase C; PPADS -pirydoxal-phosphate-6-azophenyl-2 0 , 4 0 -disulphonic acid

Amino acid variations resulting in functional and nonfunctional zebrafish P2X1 and P2X5.1 receptors

“…Support for this hypothesis comes from in vitro studies of the mouse C2C12 cell line [43] and rat satellite cells [31]. Consistent with an in vivo role of P2X receptors during muscle regeneration, the up-regulation of several P2X receptors has been observed in muscle fibers of mdx mice [32,42].…”

The role of purinergic signalling in the musculoskeletal system

“…Many studies have demonstrated the expression of multiple P2 receptor subtypes by skeletal muscle (Banachewicz et al, 2005;Cheung et al, 2003;Deli et al, 2007;Janssens et al, 1996); however, the pattern of purinergic receptor expression is strongly influenced by developmental stage and species (see (Burnstock et al, 2013)). …”

2′, 3′-O-(4-benzoylbenzoyl)–ATP-mediated calcium signaling in rat glioma C6 cells: role of the P2Y2 nucleotide receptor

“…For calcium measurements, cells were cultivated on 22-mm glass coverslips and, for the ethidium bromide uptake assay, in 24-well plates. Mouse satellite C2C12 myoblasts were cultured as described previously [18].…”