Abstract

The growth and development of embryonic skeletal muscle plays a crucial role in sheep muscle mass. But proteomic analyses for embryonic skeletal development in sheep had been little involved in the past research. In this study, we explored differential abundance proteins during embryonic skeletal muscle development by the tandem mass tags (TMT) and performed a protein profile analyses in the longissimus dorsi of Chinese merino sheep at embryonic ages Day85 (D85N), Day105 (D105N) and Day135 (D135N). 5,520 proteins in sheep embryonic skeletal muscle were identified, and 1,316 of them were differential abundance (fold change ≥1.5 and p-value < 0.05). After the KEGG enrichment analyses, these differential abundance proteins were significant enriched in the protein binding, muscle contraction and energy metabolism pathways. After validation of the protein quantification with the parallel reaction monitoring (PRM), 41% (16/39) significant abundance proteins were validated, which was similar to the results of protein quantification with TMT. All results indicated that D85N to D105N was the stage of embryonic muscle fibers proliferation, while D105N to D135N was the stage of their hypertrophy. These findings provided a deeper understanding of the function and rules of proteins in different phases of sheep embryonic skeletal muscle growth and development.

Highlights

  • The growth and development of embryonic skeletal muscle plays a crucial role in sheep muscle mass

  • The protein profile of the embryonic skeletal muscle in sheep was established by using the TMT22,23 and validated by parallel reaction monitoring (PRM) technology[24,25]

  • tandem mass tags (TMT) is widely implemented in the higher throughput proteomics, and PRM can be used to quantify protein abundance without specific antibodies and detect specific predetermined with known fragmentation characteristics more accurately in complex backgrounds than the traditional immunoassays[26,27,28,29,30]

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Summary

Introduction

The growth and development of embryonic skeletal muscle plays a crucial role in sheep muscle mass. All results indicated that D85N to D105N was the stage of embryonic muscle fibers proliferation, while D105N to D135N was the stage of their hypertrophy These findings provided a deeper understanding of the function and rules of proteins in different phases of sheep embryonic skeletal muscle growth and development. The cells are hypertrophic at later stage of growth and the number of cells are set at the phases of cell hyperplasia[6], in which some proteins and transcription factors, as regulators[7,8,9,10,11], play a cardinal role in the development process of embryonic skeletal muscle and muscle differentiation. In order to illuminate the real mechanism of embryonic skeletal muscle growth, development and maturation, a wide and deep analyses of differential abundance proteins is necessary. We constructed the protein profile of the longissimus dorsi of Chinese merino sheep at different embryonic ages and explored the function of differential abundance proteins in them

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