|
|
|
RNA-Seq explores the functional role of the fibroblast growth factor 10 gene in bovine adipocytes differentiation |
Nurgulsim Kaster1,2,a 
, Rajwali Khan1,3,a
, Ijaz Ahmad3
, Kazhgaliyev Nurlybay Zhigerbayevich4
, Imbay Seisembay2
, Akhmetbekov Nurbolat2
, Shaikenova Kymbat Hamitovna2
, Omarova Karlygash Mirambekovna2
, Makhanbetova Aizhan Bekbolatovna2
, Tlegen Garipovich Amangaliyev5
, Ateikhan Bolatbek6
, Titanov Zhanat Yeginbaevich6
, Shakoor Ahmad7
, Zan Linsen1
, Begenova Ainagul Baibolsynovna1,*
|
1College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
2Faculty of Veterinary and Livestock Technology, S. Seifullin Kazakh Agro Technical University, Astana 010000, Kazakhstan
3Department of Livestock Management, Breeding and Genetics, Faculty of Animal Husbandry and Veterinary Sciences, The University of Agriculture Peshawar, 25130, Pakistan
4Department of Livestock Management, Breeding and Genetics, Faculty of Animal Husbandry and Veterinary Sciences, The University of Agriculture Peshawar, 25130, Pakistan
5Department of Livestock Management, Breeding and Genetics, Faculty of Animal Husbandry and Veterinary Sciences, The University of Agriculture Peshawar, 25130, Pakistan
6Faculty of Agricultural Sciences, Toraighyrov University, Pavlodar 140000, Kazakhstan
7College of Veterinary Sciences, Faculty of Animal Husbandry and Veterinary Sciences, The University of Agriculture Peshawar, 25130, Pakistan |
Correspondence:
Begenova Ainagul Baibolsynovna,Email: begenova_73@mail.ru
|
Received: 17 May 2023 • Revised: 27 July 2023 • Accepted: 18 September 2023
aThese authors contributed equally to this work. |
|
|
|
| Abstract |
Objective
The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis.
Methods
The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing.
Results
The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their downregulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes.
Conclusion
Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection. |
| Keywords:
Bovine Adipocytes; FGF10 Gene; Intramuscular Fat; RNA Sequencing |
|
>
PubReader
ePub Link
Full text via DOI
Download Citation
Supplement
