Expression Characteristics Analysis of Goat (Caprahircus) Phosphodiesterase 4B (PDE4B) and Its Effect on the Differentiation ofIntramuscular Precursor Adipocytes
LI Jin-Lan1,2, SUN Shi-Yu1,2, HE Xiao-Fei3, ZHANG Li-Yi1,2, LIN Ya-Qiu1,2, LIU Wei1,2, XING Jia-Ni1,2*
1 College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China; 2 Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Ministry of Education/Key Laboratory of Sichuan Province, Southwest Minzu University, Chengdu 610041, China; 3 College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, 611130, China
Abstract：Intramuscular fat (IMF) in goat (Capra hircus) is an important factor in affecting meat quality traitssuch as flavor and juiciness. This study focused on the cloning and functional analysis of phosphodiesterases4B (PDE4B), to delineate its regulatory role in modulating the differentiation dynamics of caprine intramuscular preadipocytes. In this study, the intramuscular precursor adipocytes of JianZhou big-eared goats were used to amplify the target gene fragments by RT-PCR, and their sequences were analyzed by onlinebioinformatics tools. The expression level of PDE4B gene in different tissues and different differentiation stages of intramuscular precursor adipocytes in goats was detected using qPCR. The PDE4B overexpression plasmid (pEGFP-N1-PDE4B) was successfully engineered via molecular subcloning and subsequentlytransfected into caprine intramuscular preadipocytes. Subsequent Oil red O staining and qPCR assays were employed to assess its regulatory effects on intracellular lipid droplet deposition, coupled with transcriptional profiling of key adipogenic differentiation, triglyceride biosynthesis, and lipolytic metabolism markers. Theresults showed that the length of the cloned PDE4B gene was 1 815 bp, including the coding sequence (CDS)region of 1 629 bp, which encoded a total of 542 amino acids. Furthermore, the expression of PDE4B gene was significantly higher in goat lung than in other tissues examined (P<0.01), and reached the highest expression after 48 h induction of differentiation in intramuscular adipocytes (P<0.01). Oil red O staining and the quantitative results after PDE4B overexpression showed that there were significant increase in lipid droplet aggregation (P<0.05). The mRNA expression level of differentiation marker genes CCAAT/ enhancerbindingprotein α (C/EBPα), C/EBPβ and triglyceride synthesis marker genes activator protein 2 (AP2) and fatty acid synthase (FASN) were extremely significantly up-regulated after overexpression of PDE4B gene (P<0.01). Meanwhile, the expression of triglyceride catabolism marker genes adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) were significantly down-regulated (P<0.01). In conclusion, PDE4B gene might positively regulate the process of intramuscularprecursor adipocyte differentiation in goats, and this effect might be realized by up-regulating the expression of C/EBPα, C/EBPβ, AP2, and FASN. This study provides data support for revealing the molecular mechanism of intramuscular precursor adipocyte differentiation regulated by PDE4B in goats.
李金岚, 孙诗雨, 何晓菲, 张力懿, 林亚秋, 刘伟, 邢佳妮. 山羊PDE4B表达特性分析及其对肌内前体脂肪细胞分化的影响[J]. 农业生物技术学报, 2025, 33(6): 1312-1323.
LI Jin-Lan, SUN Shi-Yu, HE Xiao-Fei, ZHANG Li-Yi, LIN Ya-Qiu, LIU Wei, XING Jia-Ni. Expression Characteristics Analysis of Goat (Caprahircus) Phosphodiesterase 4B (PDE4B) and Its Effect on the Differentiation ofIntramuscular Precursor Adipocytes. 农业生物技术学报, 2025, 33(6): 1312-1323.
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