A new insight on copper:Promotion of collagen synthesis and myofiber growth and development in juvenile grass carp(Ctenopharyngodon idella)

Copper(Cu)is a trace element,essential for fish growth.In the current study,in addition to growth performance,we first explored the effects of Cu on collagen synthesis and myofiber growth and development in juvenile grass carp(Ctenopharyngodon idella).A total of 1080 fish(11.16±0.01 g)were randomly divided into 6 treatments(3 replicates per treatment)to receive five doses of organic Cu which were Cu citrate(CuCit)at 0.99(basal diet),2.19,4.06,6.15,and 8.07 mg/kg,and one dose o inorganic Cu(CuSO_(4)·5H_(2)O at 3.15 mg/kg),for 9 weeks.The results showed appropriate Cu level(4.06 mg kg)enhanced growth performance,improved nutritional Cu status,and downregulated Cu-transporting ATPase 1 mRNA levels in the hepatopancreas,intestine,and muscle of juvenile grass carp.Meanwhile collagen content in fish muscle was increased after Cu intake,which was probably due to the following pathways:(1)activating CTGF/TGF-β1/Smads signaling pathway to regulate collagen transcription;(2upregulating of La ribonucleoprotein domain family 6(LARP6)mRNA levels to regulate translation initiation;(3)increasing proline hydroxylase,lysine hydroxylase,and lysine oxidase activities to regulate posttranslational modifications.In addition,optimal Cu group increased myofiber diameters and the frequency of myofibers with diameter>50μm,which might be associated with upregulation of cyclin B cyclin D,cyclin E,proliferating cell nuclear antigen,myogenic determining factor(MyoD),myogenic factor 5,myogenin(MyoG),myogenic regulatory factor 4 and myosin heavy chain(MyHC)and down regulation of myostatin mRNA levels,increasing protein levels of MyoD,MyoG and MyHC in fish muscle Finally,based on percentage weight gain(PWG),serum ceruloplasmin(Cp)activity and collagen conten in fish muscle,Cu requirements were determined as 4.74,4.37 and 4.62 mg/kg diet(CuCit as Cu source of juvenile grass carp,respectively.Based on PWG and Cp activity,compared to CuSO_(4)·5H_(2)O,the efficacy of CuCit were 131.80%and 115.38%,respectively.Our findings provide new insights into Cu supple mentation to promote muscle growth in fish,and help improve the overall productivity of aquaculture.