Abstract
Whether or not the process of somitogenesis and myogenesis is affected by excessive caffeine intake still remains ambiguous. In this study, we first showed that caffeine treatment results in chest wall deformities and simultaneously reduced mRNA expressions of genes involved in myogenesis in the developing chicken embryos. We then used embryo cultures to assess in further detail how caffeine exposure affects the earliest steps of myogenesis, and we demonstrated that the caffeine treatment suppressed somitogenesis of chicken embryos by interfering with the expressions of crucial genes modulating apoptosis, proliferation, and differentiation of myogenic progenitors in differentiating somites. These phenotypes were abrogated by a retinoic acid (RA) antagonist in embryo cultures, even at low caffeine doses in C2C12 cells, implying that excess RA levels are responsible for these phenotypes in cells and possibly in vivo. These findings highlight that excessive caffeine exposure is negatively involved in regulating the development of myogenic progenitors through interfering with RA signaling. The RA somitogenesis/myogenesis pathway might be directly impacted by caffeine signaling rather than reflecting an indirect effect of the toxicity of excess caffeine dosage.
Highlights
Caffeine intake during pregnancy has been reported to be potentially teratogenic
The results demonstrated a significant suppression of cartilage fusion and muscle tissue anomaly (Masson staining, blue arrow in Supplementary Figure 2P), indicating that chest wall development is affected by directly exposing the developing chicken embryos to high doses of caffeine
Caffeine is a widely consumed psychostimulant all around the world, and its intake by adults mainly comes from coffee, FIGURE 4 | Assessment of the somitogenesis-related gene expressions following caffeine treatment. (A) ELISA data showing the concentration of retinoic acid (RA) in the control and caffeine-treated groups of HH10 chicken embryos. (B,B1) Quantitative RT-PCR data showing the mRNA expression of RALDH2 from the control and (Continued)
Summary
Caffeine intake during pregnancy has been reported to be potentially teratogenic. Maternal caffeine intake during pregnancy increases the risk of poor birth outcomes. As far as we know, muscle and cartilage cells are derived from the somites during the early stage of embryonic development (Christ and Ordahl, 1995; Jang and Baik, 2013). It still remains elusive whether or not the differentiation of somitic precursor populations of muscle cells as well as the myogenesis can be affected by caffeine exposure
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