Identification of histamine receptor subtypes in skeletal myogenesis

Abstract

To date, conventional and/or novel histamine receptors (HRs) have not been investigated in mouse skeletal myogenesis. Therefore, the present study aimed to investigate the HR-subtypes in skeletal myogenesis. The myogenesis of C2C12 skeletal myoblasts was evaluated using desmin, myogenin and myosin heavy chain (Myh) as early, intermediate and late differentiation markers, respectively. Reverse transcription-quantitative polymerase chain reaction and immunostaining were performed and the messenger RNA (mRNA) expression levels of the HR-subtypes and markers were determined. H1R mRNA was found to be highly expressed in myoblasts at day 0; however, the expression levels were reduced as differentiation progressed. By contrast, H2R mRNA expression remained constant, while H3R mRNA expression increased by 28-, 103- and 198-fold at days 2, 4 and 6 compared with the baseline level (day 0), respectively. In addition, Myh expression increased by 7,718-, 94,487- and 286,288-fold on days 2, 4 and 6 compared with the baseline expression level (day 0). Weak positive staining of the cells for H3R protein was observed on day 2, whereas highly positive staining was observed on days 4 and 6. HR expression during myogenesis was, in part, regulated by the stage of differentiation. These results along with previous findings indicated possible involvement of H1R in the regulation of progenitor cell mitogenesis and of H2R in the relaxation of acetylcholine-stimulated contraction of muscle cells, following the activation of professional histamine-producing cells, including mast cells. By contrast, H3R may participate in the regulation of specialized myocyte functions, potentially by maintaining the relaxed state under the influence of constitutive H3R activity and low histamine concentrations, locally produced/released by non-professional histamine-producing cells.

Divergent Regulation of Myotube Formation and Gene Expression by E2 and EPA during In-Vitro Differentiation of C2C12 Myoblasts

“…Morphological changes to C2C12 myoblasts cultured in control-vehicle (Con-Ve), E2, EPA and E2/EPA solutions were determined by immunofluorescence following treatment with anti-Desmin mouse antibody known to be expressed in both myoblasts and myotubes with increased expression as differentiation progresses [27]. When using immunofluorescent analyses, the staining is weak in the cytoplasm of undifferentiated myoblasts and gradually intensifies in elongated myocytes, early tubes and myotubes.…”

Activation of histamine H3 receptor decreased cytoplasmic Ca2+ imaging during electrical stimulation in the skeletal myotubes

“…The lowest (R)-α-MeHA concentration used (1 nM) was below the agonist affinity for H 3 receptor (pK i ¼ 8.4) in the brain (Chen et al, 2003 andLi et al, 2014) and had no effect on the cytoplasmic Ca 2 þ imaging under electrical stimulation in our study. On the other hand, the highest (R)-α-MeHA concentration used (100 μM) was also less active, probably because of combinatorial effects, due to nonspecific activation of other receptors (H 1 receptor or H 2 receptor) on the cell membrane (Chen et al, 2015), which counteracted the response (Leurs et al, 1998), or probably as a self-protective mechanism. The highest inhibitory effect of 1 μM (R)-α-MeHA, exerted after 10-min or 20 min treatment, was in agreement with a previous pharmacological study on histamine receptor ligands, where (R)-α-MeHA also showed the highest efficacy at 1 μM (Kottke et al, 2011).…”

Histamine deficiency delays ischaemic skeletal muscle regeneration via inducing aberrant inflammatory responses and repressing myoblast proliferation

“…21 In the early-phase of myoblast differentiation in vitro, high levels of histamine receptor expression can be detected, suggesting that histamine may participate in myoblast proliferation. 22 A previous study also reported that histamine could stimulate the expression of IGF-1 in human glioma cells. 23 These studies suggest that histamine may have an impact on the proliferation and differentiation of myoblasts and muscle regeneration.…”

“…A recent study found that histamine therapy reduces muscle injury and improves grip strength of Dmd mdx mice, but its underlying mechanism has not been reported . In the early‐phase of myoblast differentiation in vitro, high levels of histamine receptor expression can be detected, suggesting that histamine may participate in myoblast proliferation . A previous study also reported that histamine could stimulate the expression of IGF‐1 in human glioma cells .…”