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Molecular Characterizations of FAM13A and Its Functional Role in Inhibiting the Differentiation of Goat Intramuscular
Xuening Li1,2,3, Li Ran1,2,3, Yanyan Li1,2,3
1Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education, Southwest Minzu University, Chengdu 610041, China.
Abstract:
The aim of this study was to elucidate the effect of FAM13A on the differentiation of goat intramuscular precursor adipocytes and its mechanism of action. Here, we cloned the CDS region 2094 bp of the goat FAM13A gene, encoding a total of 697 amino acid residues. Functionally, overexpression of FAM13A inhibited the differentiation of goat intramuscular adipocytes with a concomitant reduction in lipid droplets, whereas interference with FAM13A expression promoted the differentiation of goat intramuscular adipocytes. To further investigate the mechanism of FAM13A inhibiting adipocyte differentiation, 104 differentially expressed genes were screened by RNA-seq, including 95 up-regulated genes and 9 down-regulated genes. KEGG analysis found that the RIG-I receptor signaling pathway, NOD receptor signaling pathway and toll-like receptor signaling pathway may affect adipogenesis. We selected the RIG-I receptor signaling pathway enriched with more differential genes as a potential adipocyte differentiation signaling pathway for verification. Convincingly, the RIG-I like receptor signaling pathway inhibitor (HY-P1934A) blocked this pathway to save the phenotype observed in intramuscular adipocyte with FAM13A overexpression. Finally, the upstream miRNA of FAM13A was predicted, and the targeted inhibition of miR-21-5p on the expression of FAM13A gene was confirmed. In this study, it was found that FAM13A inhibited the differentiation of goat intramuscular adipocytes through the RIG-I receptor signaling pathway, and the upstream miRNA of FAM13A (miR-21-5p) promoted the differentiation of goat intramuscular adipocytes. This work extends the genetic regulatory network of IMF deposits and provides theoretical support for improving human health and meat quality from the perspective of IMF deposits.
Insights
FAM13A protein inhibits goat adipocyte differentiation via the RIG-I signaling pathway. MiR-21-5p, a key upstream regulator, promotes adipocyte differentiation, offering insights into intramuscular fat deposition and meat quality.
Area of Science:
- Animal Science
- Molecular Biology
- Genetics
Background:
- Intramuscular fat (IMF) deposition is crucial for meat quality and human health.
- Understanding the genetic regulation of adipocyte differentiation is essential for optimizing meat production.
Purpose of the Study:
- To investigate the role of FAM13A in goat intramuscular adipocyte differentiation.
- To elucidate the underlying molecular mechanisms, including signaling pathways and upstream regulators.
Main Methods:
- Cloning of the goat FAM13A gene.
- Overexpression and interference assays of FAM13A in goat adipocytes.
- RNA sequencing (RNA-seq) and KEGG pathway analysis.
- Inhibition of the RIG-I receptor signaling pathway.
- miRNA prediction and validation.
Main Results:
- FAM13A overexpression inhibited adipocyte differentiation and reduced lipid droplets.
- FAM13A interference promoted adipocyte differentiation.
- RNA-seq identified the RIG-I receptor signaling pathway as a key mediator.
- Inhibition of RIG-I signaling rescued the phenotype of FAM13A overexpression.
- miR-21-5p was confirmed as an upstream regulator that targets FAM13A.
Conclusions:
- FAM13A negatively regulates goat intramuscular adipocyte differentiation through the RIG-I receptor signaling pathway.
- miR-21-5p promotes adipocyte differentiation by inhibiting FAM13A.
- This study expands the understanding of the genetic regulatory network of IMF deposition, providing theoretical support for improving meat quality.
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