The Wnt/β-catenin signaling pathway regulates cell proliferation and differentiation to determine cell fate during embryogenesis. Lithium chloride (LiCl) is known to activate canonical Wnt signaling by inhibiting glycogen synthetase kinase-3β and consequently stabilizing free cytosolic β-catenin. To understand the role of the Wnt/β-catenin pathway in the regulation of porcine myoblast differentiation, we studied the effects of LiCl on cultured porcine myoblasts and β-catenin expression. A supplementation of 25 mM LiCl induced myoblast differentiation into myotubes over 3 days of culture. By semi-quantitative RT-PCR analyses, levels of mRNA encoding MyoD, Myogenin, Myf5 and several Wnt-responsive genes in the cultured myoblast cells were significantly increased after LiCl treatment. Using Western blotting and immunofluorescence analysis, we found that the protein levels of β-catenin were consistently increased by LiCl. Meanwhile, phosphorylated GSK-3β at Ser9 levels were also increased as an indicator of GSK-3β inactivation. Additionally, the nuclear staining of endogenous β-catenin was also significantly increased in porcine myoblasts 48 h after LiCl treatment. These results provided additional evidence that Wnt/β-catenin is a significant pathway that regulates myogenic differentiation. An enhanced level of β-catenin plays a positive role in porcine myoblast differentiation.
As the fastest growing food production sector in the world, aquaculture may become an important source of nitrous oxide (N2O)—a potent greenhouse gas and the dominant source of ozone-depleting substances in the stratosphere. China is the largest aquaculture producer globally; however, the magnitude of N2O emission from Chinese aquaculture systems (CASs) has not yet been extensively investigated. Here, we quantified N2O emission from the CASs since the Reform and Opening-up (1979–2019) at the species-, provincial-, and national-levels using annual aquaculture production data, based on nitrogen (N) levels in feed type, feed amount, feed conversion ratio, and emission factor (EF). Our estimate indicates that over the past 41 years, N2O emission from CASs has increased approximately 25 times from 0.67 ± 0.04 GgN in 1979 to 16.69 ± 0.31 GgN in 2019. Freshwater fish farming, primarily in two provinces, namely, Guangdong and Hubei, where intensive freshwater fish farming has been adopted in the past decades, accounted for approximately 89% of this emission increase. We also calculated the EF for each species, ranging from 0.79 ± 0.23 g N2O kg−1 animal to 2.41 ± 0.14 g N2O kg−1 animal. The results of this study suggest that selecting low-EF species and improving feed use efficiency can help reduce aquaculture N2O emission for building a climate-resilient sustainable aquaculture.
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