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Screening of key miRNAs related with the differentiation of subcutaneous adipocytes and the validation of miR-133a-3p functional significance in goats.

Objective
Adipocyte differentiation is regulated by a variety of functional genes and non-coding RNAs. However, the role of miRNAs in lipid deposition of goat white adipose tissue is still unclear. Therefore, this study revealed the miRNA expression profile in goat subcutaneous adipocytes by sRNA-seq.


Methods
The miRNA expressed in goat subcutaneous preadipocytes and the mature adipocytes were sequenced by sRNA-seq. The differentially expressed miRNAs (DEm) were screened and GO and KEGG analyses were performed. Gain-of-function and loss-of-function combined with Oil Red O staining, Bodipy staining, and qPCR were utilized to determine the effect of miR-133a-3p on adipocyte differentiation.


Results
A total of 218 DEm were screened out. The target genes of these DEm were significantly enriched in GO items such as biological regulation and in KEGG terms such as FAK signaling pathway and MAPK signaling pathway. qPCR verified that the expression trend of miRNA was consistent with miRNA-seq. The gain-of-function or loss-of-function of miR-133a-3p showed that it promoted or inhibited the accumulation of lipid droplets, and C/EBPα and C/EBPβ were extremely significantly up-regulated or down-regulated respectively (P<0.01), the loss-of-function also led to a significant down-regulation of PPARγ (P<0.01).


Conclusion
This study successfully identified miRNAs expression patterns in goat subcutaneous adipocytes, and functional identification indicates that miR-133a-3p is a positive regulator of the differentiation process of goat subcutaneous adipocytes. Our results lay the foundation for the molecular mechanism of lipid deposition in meat-source goats from the perspective of miRNA.