Molecular Characterization of LKB1 of Triploid Crucian Carp and Its Regulation on Muscle Growth and Quality

SIMPLE SUMMARY: Liver Kinase B1 (LKB1) can regulate energy metabolism and skeletal muscle growth. In the study, we cloned LKB1 cDNA and assessed LKB1 expression in triploid crucian carp (Carassius auratus), the molecular regulation mechanisms of LKB1 involved in muscle growth and quality. In conclusion, the LKB1 amino acid sequence of the triploid crucian carp had a high sequence identity with carp (Cyprinus carpio). Additionally, activated LKB1 can promote the expression of most myogenic regulatory factors, muscle fibers development, the formation of inosine monophosphate (IMP), which is a flavor substance of muscle, and improve meat quality by increasing elasticity and chewiness. Inhibiting LKB1 can reverse the trend. Moreover, with increasing lysine-glutamate dipeptide concentrations in the feed, the expression of LKB1 gradually increased and was highest when dipeptide concentration was 1.6%. ABSTRACT: Liver Kinase B1 (LKB1) is a serine/threonine kinase that can regulate energy metabolism and skeletal muscle growth. In the present study, LKB1 cDNA of triploid crucian carp (Carassius auratus) was cloned. The cDNA contains a complete open reading frame (ORF), with a length of 1326 bp, encoding 442 amino acids. Phylogenetic tree analysis showed that the LKB1 amino acid sequence of the triploid crucian carp had a high sequence similarity and identity with carp (Cyprinus carpio). Tissue expression analysis revealed that LKB1 was widely expressed in various tissues. LKB1 expressions in the brain were highest, followed by kidney and muscle. In the short-term LKB1 activator and inhibitor injection experiment, when LKB1 was activated for 72 h, expressions of myogenic differentiation (MyoD), muscle regulatory factor (MRF4), myogenic factor (MyoG) and myostatin 1 (MSTN1) were markedly elevated and the content of inosine monophosphate (IMP) in muscle was significantly increased. When LKB1 was inhibited for 72 h, expressions of MyoD, MyoG, MRF4 and MSTN1 were markedly decreased. The long-term injection experiment of the LKB1 activator revealed that, when LKB1 was activated for 15 days, its muscle fibers were significantly larger and tighter than the control group. In texture profile analysis, it showed smaller hardness and adhesion, greater elasticity and chewiness. Contrastingly, when LKB1 was inhibited for 9 days, its muscle fibers were significantly smaller, while the gap between muscle fibers was significantly larger. Texture profile analysis showed that adhesion was significantly higher than the control group. A feeding trial on triploid crucian carp showed that with dietary lysine-glutamate dipeptide concentration increasing, the expression of the LKB1 gene gradually increased and was highest when dipeptide concentration was 1.6%. These findings may provide new insights into the effects of LKB1 on fish skeletal muscle growth and muscle quality, and will provide a potential application value in improvement of aquaculture feed formula.