Scientia Agricultura Sinica ›› 2025, Vol. 58 ›› Issue (15): 3134-3144.doi: 10.3864/j.issn.0578-1752.2025.15.014

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles     Next Articles

Expression Pattern of gga-miR-30a-5p and Its Regulation of Abdominal Fat and Intramuscular Fat Deposition in Chicken

HUANG HuaYun(), LIU Xing, WANG QianBao, LI RuiRui, YANG MiaoMiao, LI ChunMiao, WU ZhaoLin, KONG LingLin, ZHAO ZhenHua*()   

  1. Jiangsu Institute of Poultry Science, Yangzhou 225125, Jiangsu
  • Received:2025-02-12 Accepted:2025-06-07 Online:2025-08-01 Published:2025-07-30
  • Contact: ZHAO ZhenHua

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Abstract:

【Objective】 miR-30a-5p is a member of the miR-30 family, and its role in chicken fat deposition has not been reported. This study focused on the role of gga-miR-30a-5p in chicken abdominal fat and intramuscular fat deposition, which could lay a foundation for the analysis of the mechanism of chicken abdominal fat and intramuscular fat deposition. 【Method】 This study used high-quality yellow-feathered broilers (dwarf line, S3) and recessive white-feathered chickens (RR) as experimental subjects. Quantitative real-time PCR (qPCR) was employed to detect the expression changes of gga-miR-30a-5p in abdominal fat, liver, and leg muscle tissues at 0, 2, 4, 8, 14, and 16 weeks (0, 2, 4, 8, 14, and 16W), respectively, as well as during the proliferation and differentiation stages of abdominal and intramuscular adipocytes. Abdominal and intramuscular adipocytes were transfected with gga-miR-30a-5p mimics and inhibitors, and qPCR was used to measure the expression changes of gga-miR-30a-5p. Oil Red O staining and isopropanol extraction were performed to assess lipid droplet deposition after transfection. Bioinformatics analysis was conducted to predict the target genes of gga-miR-30a-5p. 【Result】 The expression of gga-miR-30a-5p in different tissues (abdominal fat, liver, and leg muscle) showed significant breed-specific differences (P<0.05). In abdominal fat tissue, the expression of gga-miR-30a-5p in recessive white-feathered chickens at 0W was significantly lower than that in S3 chickens (P<0.05). In both S3 and RR chickens, gga-miR-30a-5p expression in abdominal fat was the highest at 0W, significantly higher than that at other weeks (P<0.05), with no significant differences among other weeks (P>0.05). In liver tissue, gga-miR-30a-5p expression in RR chickens at 16W was significantly lower than that in S3 chickens (P<0.05). In S3 chickens, gga-miR-30a-5p expression in liver tissue at 16W was significantly higher than that at other weeks (P<0.05), while in RR chickens, its expression at 16W was significantly higher than that at 0 and 8W (P<0.05). In leg muscle tissue, gga-miR-30a-5p expression in RR chickens at 16W was significantly lower than that in S3 chickens (P<0.05). In S3 chickens, gga-miR-30a-5p expression in leg muscle was the lowest at 14W, significantly lower than that at 2, 8, and 16W (P<0.05), whereas no significant differences were observed among weeks in RR chickens (P>0.05). In abdominal adipocytes, gga-miR-30a-5p expression during the proliferation stage was significantly lower than that at differentiation days 4 and 6 (P<0.05). In intramuscular adipocytes, no significant difference was observed between the proliferation stage and differentiation day 1 (P>0.05), but expression at differentiation days 4 and 6 was significantly higher than during proliferation (P<0.05), gradually increasing with prolonged differentiation. After transfection with gga-miR-30a-5p mimics or inhibitors for 24 hours, gga-miR-30a-5p expression in abdominal adipocytes significantly increased or decreased, confirming successful transfection. Transfection with gga-miR-30a-5p mimics for 3 days significantly enhanced lipid droplet deposition in abdominal adipocytes, whereas transfection with inhibitors significantly reduced deposition (P<0.05). Similarly, in intramuscular adipocytes, transfection with mimics or inhibitors for 24 hours significantly altered gga-miR-30a-5p expression. Mimics transfection for 3 days significantly increased lipid droplet deposition, while inhibitor transfection significantly decreased it (P<0.05). Target gene prediction via GO, Pathway, and protein-protein interaction analyses suggested that UBE2I, UBE3C, CUL2, SOSC3, and RUNX1 were potential target genes of gga-miR-30a-5p in regulating fat deposition.【Conclusion】 gga-miR-30a-5p exhibited significant breed-specific expression differences across tissues, which promoted abdominal and intramuscular fat deposition, with UBE2I, UBE3C, CUL2, SOSC3, and RUNX1 identified as key candidate target genes.

Key words: chicken, gga-miR-30a-5p, abdominal fat, intramuscular fat

Fig. 1

Change of gga-miR-30a-5p expression in abdominal adipose tissue in S3 strains and recessive white feathered chickens S3: S3 Strain; RR: Recessive white feather chickens. Different lowercase letters indicate significant differences between different periods of the same breed (P<0.05), and different capital letters indicate significant differences between breeds (P<0.05). * P<0.05. The same as below"

Fig. 2

Change of gga-miR-30a-5p expression of liver tissue in S3 strain and recessive white feathered chickens"

Fig. 3

Change of gga-miR-30a-5p expression of leg muscle tissue in S3 strain and recessive white feathered chickens"

Fig. 4

Change of gga-miR-30a-5p expression in abdominal and intramuscular adipocytes A: Abdominal adipocytes; B: Intramuscular adipocytes. Different superscripts indicate significant differences among stages (P<0.05)"

Fig. 5

Change of gga-miR-30a-5p expression after gga-miR-30a-5p mimics/inhibitor transfection A: mimics group；B：inhibitor group. *** indicates very significant difference from control group（P<0.001）,** indicates very significant difference from control group（P<0.01）. The same as below"

Fig. 6

Changes in lipid droplet deposition in abdominal adipocytes and in intramuscular adipocytes after transfection with gga-miR-30a-5p mimics/inhibitor A: Abdominal adipocytes transfected with mimics; B: Abdominal adipocytes transfected with inhibitor; C: Intramuscular adipocytes transfected with mimics; D: Intramuscular adipocytes transfected with inhibitor"

Fig. 7

Predicted target genes, GO entries, and KEGG pathways of gga-miR-30a-5p A: Predicted target gene of gga-miR-30a-5p; B: GO entries significantly enriched for the target gene; C: KEGG pathway significantly enriched for the target gene"

Table 1

Significantly enriched pathways for predicted target genes"

通路 Pathway P值P value 富集基因Enriched gene
MAPK信号通路
MAPK signaling pathway		 0.0001 BDNF, NFATC3, CACNA1D, CACNA1C, CRKL, RAP1B, CACNB2, PPM1A, PPP3CA, MAPK8, PPP3CB, PPP3R1, GDNF, TAOK1, RASA1, RASA2, NF1, RAPGEF2, KRAS, SOS1, MAP4K4, MAP3K5
泛素介导的蛋白水解
Ubiquitin mediated proteolysis		 0.0012 UBE2F, UBE2I, UBE3C, CUL2, UBE2D3, NEDD4L, WWP1, CBLB, UBE2J1, SOCS3, HERC2, NEDD4, TRIP12
多聚酶抑制复合体
Polycomb repressive complex		 0.0025 CBX8, PCGF5, EED, ASXL3, PCGF3, UBE2D3, LCOR, BCOR, YAF2
紧密连接
Tight junction		 0.0337 RAP2C, MAPK8, NEDD4, ACTN1, NEDD4L, RAPGEF2, CACNA1D, AMOTL2, RAB8A, MAP3K5, RUNX1

Fig. 8

Interaction map of target genes"

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