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. 2020 Jun 26;40(6):BSR20193796.
doi: 10.1042/BSR20193796.

Potential role of miR-155-5p in fat deposition and skeletal muscle development of chicken

Sifan Xu  1 , Yang Chang  1 , Guanxian Wu  1 , Wanting Zhang  1 , Chaolai Man  1
Affiliations

Potential role of miR-155-5p in fat deposition and skeletal muscle development of chicken

Sifan Xu et al. Biosci Rep. .

Abstract

miR-155 has multiple functions in many physiological and pathological processes. However, little is known about the expression characteristics of avian miR-155. In the present study, partial pri-miR-155 sequences were cloned from AA+ broiler, Sanhuang broiler and Hy-Line Brown layer, respectively. Stem-loop qRT-PCR was performed to detect the miR-155-5p spatiotemporal expression profiles of each chicken breed, and the target genes of miR-155-5p were predicted in Gene Oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The results showed that the partial pri-miR-155 sequences of different breeds of chicken were high conserved. The expression patterns of miR-155-5p between broiler and layer were basically similar, and miR-155-5p was expressed highly in immune related tissues (spleen, thymus and bursa). In the same old chicken (14 days old), miR-155-5p expression activity of fat tissue all had higher level in the three chicken breeds, but the expression activities in skeletal muscle of broilers were significantly lower than that of layer (P<0.05). In different development stages of Hy-Line Brown layer, miR-155-5p expression activities in skeletal muscle of 14-day-old and 10-month-old layers were significantly lower than that of 24-month-old layer (P<0.05). Fat related target genes (ACOX1, ACOT7, FADS1, SCD and HSD17B12) and skeletal muscle related target genes (CCNT2, DMD, CFL2, MAPK14, FLNB, ZBTB18 and CDK5) of miR-155-5p were predicted, respectively. The results indicate that miR-155-5p may be an important factor inhibiting the fat deposition and skeletal muscle development in chicken.

Keywords: chicken; clone; expression; fat; miR-155-5p; skeletal muscle.

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Conflict of interest statement

The authors declare that there are no competing interests associated with the manuscript.

Figures

Figure 1
Figure 1. Partial pri-miR-155 sequence of chicken
Nucleotide homology analysis of partial pri-miR-155 sequence of chicken using DNAMAN software (http://www.lynnon.com). The sequence of pre-miR-155 is shaded.
Figure 2
Figure 2. Homology and evolutionary analysis of multiple sequences of pre-miR-155
(A) Homology tree of multiple sequences of pre-miR-155. Homology tree of pre-miR-155 based on the nucleotide sequences. Analysis was done using the DNAMAN software (http://www.lynnon.com). The percentages on the branches represented homology. Interval range of scale corresponds to homology value of homologous tree. (B) Phylogenetic tree of multiple sequences of pre-miR-155. The tree was obtained by boot strap analysis with the neighbor-joining method; numbers on the branches represent bootstrap values for 1000 replications. aca-mir-155 (Anolis carolinensis, miRBase: MI0018764); bta-mir-155 (Bos taurus, miRBase: MI0009752); ccr-mir-155 (Cyprinus carpio, miRBase: MI0023336); cfa-mir-155 (Canis familiaris, miRBase: MI0008078); cgr-mir-155 (Cricetulus griseus, miRBase: MI0020415); chi-mir-155 (Capra hircus miRBase: MI0030643); dre-mir-155 (Danio rerio, miRBase: MI0002023); eca-mir-155 (Equus caballus, miRBase: MI0012927); efu-mir-155 (Eptesicus fuscus, miRBase: MI0028748); gga-mir-155 (Gallus gallus, miRBase: MI0001176); ggo-mir-155 (Gorilla gorilla, miRBase: MI0020768); hsa-mir-155 (Homo sapiens, miRBase: MI0000681); ipu-mir-155 (Ictalurus punctatus, miRBase: MI0024518); mml-mir-155 (Macaca mulatta, miRBase: MI0007645); mmu-mir-155 (Mus musculus, iRBase: MI0000177); oan-mir-155 (Ornithorhynchus anatinus, miRBase: MI0006775); ppy-mir-155 (Pongo pygmaeus, miRBase: MI0014843); ptr-mir-155 (Pan troglodytes, miRBase: MI0008554); rno-mir-155 (Rattus norvegicus, miRBase: MI0025509); ssa-mir-155-1 (Salmo salar, miRBase: MI0026520); ssc-mir-155 (Sus scrofa, miRBase: MI0015907); tch-mir-155 (Tupaia chinensis, miRBase: MI0031131); tgu-mir-155 (Taeniopygia guttata, miRBase: MI0013842); xtr-mir-155 (Xenopus tropicalis, miRBase: MI0004848).
Figure 3
Figure 3. Expression patterns of miR-155-5p analyzed by stem-loop qRT-PCR
(A) Expression distributions of miR-155-5p in three breeds of 14-day-old chicken. (B) Expression distributions of miR-155-5p in different developmental stages of Hy-Line Brown layer (14 days old, 10 months old and 24 months old). (C) Expression levels of miR-155-5p in skeletal muscle of three breeds of 14 days old chicken. (D) Expression levels of miR-155-5p in skeletal muscle of different developmental stages of Hy-Line Brown layer. 1 heart, 2 liver, 3 spleen, 4 lung, 5 kidney, 6 brain, 7 skeletal muscle, 8 muscular stomach, 9 thymus, 10 skin, 11 small intestine, 12 large intestine, 13 glandular stomach, 14 fat, 15 blood, 16 bursa. The internal reference gene is U6. The data are indicated as mean ± SD. Different letters (a−f) among different columns represent significant differences (P<0.05), while the same letters among different columns represent no significant differences.
Figure 4
Figure 4. Target gene analysis of miR-155-5p
(A) MiR-155-5p target genes associated with biosynthesis of unsaturated fatty acids (hsa01040). (B) MiR-155-5p target genes associated with skeletal muscle development (GO: 0007519). Acquisition of miR-155-5p target genes that had been experimentally verified using miRWalk 2.0 (http://zmf.umm.uni-heidelberg.de/apps/zmf/mirwalk2/). Then, Gene Oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the target genes (P<0.05) by DAVID v6.8 (https://david.ncifcrf.gov/).
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