简州大耳羊肌内脂肪细胞成脂分化差异表达基因的筛选与鉴定

doi:10.11843/j.issn.0366-6964.2020.07.006

Abstract

Abstract: This experiment aimed to obtain the differentially expressed genes before and after the adipogenic differentiation of Jianzhou Da'er goat intramuscular preadipocytes by the RNA-seq technology, and acquire the related signaling pathways and key candidate functional genes by bioinformatics analysis. The 7-day-old healthy male Jianzhou Da'er goat was used (n=3) and their intramuscular preadipocytes were isolated by collagenase digestion. Illumina platform was used to carry out the high-throughput sequencing to two groups of cDNA samples (n=3) of Jianzhou Da'er goat intramuscular preadipocytes and intramuscular adipocytes induced for 5 day, respectively. The differentially expressed genes were obtained according to the thresholds of|log₂fold change|>0 and P<0.05. The differentially expressed genes were enriched to GO terms and KEGG pathways using the clusterProfiler R. Ultimately, the qRT-PCR technology was performed to detect the change of expression levels of candidate functional genes before and after differentiation of preadipocytes. The results showed that there were 7 916 differentially expressed genes in the RNA-seq results during the adipogenic differentiation of Jianzhou Da'er goat intramuscular preadipocytes, including 4 143 up-regulated genes that were enriched to 305 KEGG pathways, including oxidative phosphorylation, ribosome synthesis, and citrate cycle pathways, etc., and 3 773 down-regulated genes that were enriched to 303 KEGG pathways, including thyroxine signaling pathways, and so on; In the results of GO function enrichment analysis, the biological process was 52.8% of GO terms, the cellular component was 13.4%, and the molecular function was 33.8%. The qRT-PCR results showed that the trends of UCP3, ACACB, ACOT11, ACOX3, APOA1, and WISP2 expression during the adipogenic differentiation of intramuscular preadipocytes were consistent with the results of RNA-seq, which indicated that these genes were the suitable candidate functional genes for further research. This study obtained the differentially expressed genes during the adipogenic differentiation of Jianzhou Da'er goat intramuscular adipocytes and verified 6 candidate functional genes. The results of this study provided the basic data and systematic information for elucidating the molecular regulatory networks during the adipogenic differentiation of meat goat intramuscular adipocytes.

Key words: Jianzhou Da'er goat, intramuscular preadipocytes, RNA-seq technology, differentially expressed genes, induced differentiation

CLC Number:

S827.2

XIE Guangjie, WANG Yong, XU Qing, ZHU Jiangjiang, LIN Yaqiu. Selection and Validation of the Differentially Expressed Genes during the Adipogenic Differentiation of Jianzhou Da'er Goat Intramuscular Adipocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(7): 1525-1536.

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Selection and Validation of the Differentially Expressed Genes during the Adipogenic Differentiation of Jianzhou Da'er Goat Intramuscular Adipocytes

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