The circadian rhythm regulates branched-chain amino acids metabolism in fast muscle of Chinese perch (Siniperca chuatsi) during short-term fasting by Clock-KLF15-Bcat2 pathway

As an internal time keeping mechanism, circadian rhythm plays crucial role in maintaining homeostasis when in response to nutrition change, meanwhile, branched-chain amino acids (BCAAs) in skeletal muscle play an important role in preserving energy homeostasis during fasting. Previous results from our lab suggested that fasting can influence peripheral circadian rhythm and BCAAs metabolism in fish, but the relationship between circadian rhythm and BCAAs metabolism, and whether circadian rhythm regulates BCAAs metabolism to maintain physiological homeostasis during fasting remains unclear. This study shows that the expression of 15 core clock genes, as well as KLF15 and Bcat2 is highly responsive to short-term fasting in fast muscle of Siniperca chuatsi, and the correlation coefficient between Clock and KLF15 expression is enhanced after fasting treatment. Furthermore, we demonstrate that the transcriptional expression of KLF15 is regulated by Clock, and the transcriptional expression of Bcat2 is regulated by KLF15 by using dual-luciferase reporter gene assay and Vivo-Morpholinos mediated gene knockdown technique. Therefore, fasting imposes a dynamic coordination of transcription between the circadian rhythm and BCAAs metabolic pathways. The findings highlight the interaction between circadian rhythm and BCAAs metabolism, and suggest that fasting induces a switch in KLF15 expression through affecting the rhythmic expression of Clock, and then KLF15 promotes the transcription of Bcat2 to enhance the metabolism of BCAAs, thus maintaining energy homeostasis and providing energy for skeletal muscle as well as other tissues.