Comparative Analysis Of Proteomics And MiRNA In The Fast And Slow Muscle Of Siniperca Chuatsi

Fish meat has been one of the important sources of daily intake of meat protein,and the status of aquaculture in China’s agriculture is also increasing.The skeletal muscle of Siniperca chuatsi constitutes the main component of its body,and can be divided into fast muscle fiber and slow muscle fiber which are great differences in the physiological characteristics and metabolic modes of these two different muscle fiber types.The different characteristics and types of fish muscle fibers are one of the important factors affecting the quality of fish meat,and excellent meat quality of Siniperca chuatsi is closely related to the unique properties of its different skeletal muscle fibers.In order to fully understand the traits of different types of skeletal muscle fibers in Siniperca chuatsi,we combined the proteomics and miRNAomics to comprehensively analyze and compare differentially expressed proteins,miRNAs and their related pathways in fast and slow muscles for further analysis which to provide theoretical basis for studies on the growth and development of fish muscles.The results of this research are as follows:1.Analysis of fast and slow muscle protein group of mandarin fish.To gain insights into protein differences between fast and slow muscles,we used i TRAQ-based quantitative proteomics to analyze proteome differences in fast muscle and slow muscle fibers,yielded a total of 328,763 spectra,identified 9711 peptides and 2102 different proteins.Among of them,499 proteins were differentially expressed in the fast and slow muscles of carp.Compared with slow muscles,there were 99 differentially upregulated proteins and 400 differentially expressed proteins(Fold chang ≥1.2 or ≤0.83).Q-value ≤0.05).Among these differentially expressed proteins,we also screened 24 proteins related to muscle fiber traits.We also performed GO,COG and KEGG pathway assays for all identified and differentially expressed proteins.2.Analysis of miRNAs expression profiles of fast and slow muscles of mandarin fish.Through chip data analysis,59 miRNAs were significantly different in fast and slow muscles(p<0.05),and 32 miRNAs were significantly up-regulated and 27 significantly down-regulated.There are 7 miRNAs with high abundance in fast muscle and 4 were in slow muscle and we screened 19 muscle-associated miRNAs and verified the expression levels of miR-103 and miR-144 in the two types of muscle tissues by SYBR Green q PCR.The results were consistent with the microarray,miR-103 has a high expression in the slow muscle,but the expression of miR-144 is high in the fast muscle.3.Prediction and validation of miR103 and miR144 target genes.We found that miR-103 and miR-144 were able to find predicted target sites in the Smy D1 a and Smy D1 b genes,respectively,indicating that Sym D1 a and Smy D1 b may be miR-103 and miR-144 target genes.Double luciferase assays verified that Sym D1 a and Smy D1 b are miR-103 and miR-144 target genes.Maintenance and function of muscles require the interaction of transcription regulators,structural proteins,chaperones,and micro RNAs.Based on the published research results and the data we have obtained,we have constructed a predictive regulatory network that shows the interaction between Smy D1 and other regulatory factors and structural proteins as well as fast and slow muscle fiber formation and functional micro RNAs.Among them,miR-103 and miR-144 mainly regulate their target genes Smy D1 a and Smy D1 b,and further act on their downstream functional genes Unc45 b and Hsp90a1 to achieve the effect of regulating muscle growth and differentiation.The specific regulation mechanism of miR-103 and miR-144 remains to be further studied.