MiRNAs and mRNAs Analysis during Abdominal Preadipocyte Differentiation in Chickens.

Xiangfei Ma,Zhuanjian Li,Wenting Li,Donghua Li,Junwei Sun,Shuaipeng Zhu,Yadong Tian,Guirong Sun,Xiangtao Kang,Zhenwei Du

doi:10.3390/ani10030468

Abstract

Simple SummaryWe sequenced the miRNAs and mRNAs of preabdominal fat cells and differentiated adipocytes, and target genes of miRNA combined with mRNA transcriptome data jointly. We found that the MAPK signal pathway, insulin signal pathway, fatty acid metabolism, ECM( extracellular matrix)–receptor interaction, and other signal pathways were involved in the differentiation of preabdominal fat cells. In addition, we found that some miRNAs–mRNAs combinations were strongly related to the differentiation of fat cells (miR-214−ACSBG2, NFKB2, CAMK2A, ACLY, CCND3, PLK3, ITGB2; miR-148a-5p−ROCK2; miR-10a-5p−ELOVL5; miR-146b-5p−LAMA4; miR-6615-5p−FLNB; miR-1774−COL6A1). Our findings provide important resources for the study of adipocyte differentiation.The excessive deposition of abdominal fat has become an important factor in restricting the production efficiency of chickens, so reducing abdominal fat deposition is important for improving growth rate. It has been proven that miRNAs play an important role in regulating many physiological processes of organisms. In this study, we constructed a model of adipogenesis by isolating preadipocytes (Ab-Pre) derived from abdominal adipose tissue and differentiated adipocytes (Ab-Ad) in vitro. Deep sequencing of miRNAs and mRNAs expressed in Ab-Pre and Ab-Ad groups was conducted to explore the effect of miRNAs and mRNAs on fat deposition. We identified 80 differentially expressed miRNAs (DEMs) candidates, 58 of which were up-regulated and 22 down-regulated. Furthermore, six miRNAs and six mRNAs were verified by qRT-PCR, and the results showed that the expression of the DEMs and differentially expressed genes (DEGs) in the two groups was consistent with our sequencing results. When target genes of miRNA were combined with mRNA transcriptome data, a total of 891 intersection genes were obtained, we predicted the signal pathways of cross genes enrichment to the MAPK signal pathway, insulin signal pathway, fatty acid metabolism, and ECM–receptor interaction. Meanwhile, we constructed miRNA and negatively correlated mRNA target networks, including 12 miRNA–mRNAs pairs, which showed a strong association with the abdominal adipocyte differentiation (miR-214−ACSBG2, NFKB2, CAMK2A, ACLY, CCND3, PLK3, ITGB2; miR-148a-5p−ROCK2; miR-10a-5p−ELOVL5; miR-146b-5p−LAMA4; miR-6615-5p−FLNB; miR-1774−COL6A1). Overall, these findings provide a background for further research on lipid metabolism. Thus, we can better understand the molecular genetic mechanism of chicken abdominal fat deposition.

Highlights

With the development of intensive feeding, the daily growth rate and bodyweight of chickens were tended to be selected, but the accumulation of high abdominal fat is still an obstacle [1,2].In poultry, excessive abdominal fat deposition increases the cost of feeding and negatively affects the appearance of products, and poses a threat to the health of consumers [3,4]
Some studies have shown that miRNAs play an important role in regulating adipocyte differentiation and lipid metabolism [11,12]
Regulation of miRNAs on human adipocyte differentiation has been reported. miR-361-5p and miR-574-5p may be associated with hypertrophy of human white adipose tissue (WAT) by affecting the expression of EBF1 [15]

Summary

Introduction

With the development of intensive feeding, the daily growth rate and bodyweight of chickens were tended to be selected, but the accumulation of high abdominal fat is still an obstacle [1,2].In poultry, excessive abdominal fat deposition increases the cost of feeding and negatively affects the appearance of products, and poses a threat to the health of consumers [3,4]. Our previous studies have shown that abdominal preadipocytes have higher adipogenic differentiation ability than intramuscular preadipocytes [5], so reducing abdominal fat deposition is of great significance to the poultry industry. MiRNAs have the function of regulating gene expression at translation level or transcriptional level. The regulation of mature miRNAs on target genes depends on the complementarity between the seed sequence of miRNAs and the 30 UTR regulatory region of the target gene, leading to translation inhibition or gene splicing [6,7,8]. Some studies have shown that miRNAs play an important role in regulating adipocyte differentiation and lipid metabolism [11,12]. Regulation of miRNAs on human adipocyte differentiation has been reported. There are few studies on miRNA of chicken abdominal adipocyte

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