JIA-2021-1043 转录组分析解析转人FST基因猪表型变化的分子机制

doi:10.1016/j.jia.2022.07.014

Journal of Integrative Agriculture ›› 2022, Vol. 21 ›› Issue (9): 2675-2690.DOI: 10.1016/j.jia.2022.07.014

所属专题：动物科学合辑Animal Science

JIA-2021-1043 转录组分析解析转人FST基因猪表型变化的分子机制

收稿日期:2021-06-16 接受日期:2022-03-07 出版日期:2022-09-01 发布日期:2022-03-07

Transcriptomic analysis elucidates the enhanced skeletal muscle mass, reduced fat accumulation, and metabolically benign liver in human follistatin-344 transgenic pigs

LONG Ke-ren^1*, LI Xiao-kai^1*, ZHANG Ruo-wei^1*, GU Yi-ren^2*, DU Min-jie⁵, XING Xiang-yang⁵, DU Jia-xiang⁵, MAI Miao-miao¹, WANG Jing¹, JIN Long¹, TANG Qian-zi¹, HU Si-lu¹, MA Ji-deng¹, WANG Xun¹, PAN Deng-ke^{3, 4}, LI Ming-zhou¹

¹ Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, P.R.China
² Animal Breeding and Genetics Key Laboratory of Sichuan Province, Pig Science Institute, Sichuan Animal Science Academy, Chengdu 610066, P.R.China
³ Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
⁴ Institute of Organ Transplantation, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Chengdu 610072, P.R.China
⁵ Chengdu Clonorgan Biotechnology Co., Ltd., Chengdu 610041, P.R.China

Received:2021-06-16 Accepted:2022-03-07 Online:2022-09-01 Published:2022-03-07
About author:LONG Ke-ren, E-mail: keren.long@sicau.edu.cn; LI Xiao-kai, E-mail: xiaokai_li2017@163.com; ZHANG Ruo-wei, E-mail: ruowei2021@163.com; GU Yi-ren, E-mail: guyiren1128@163.com; Correspondence LI Ming-zhou, E-mail: mingzhou.li@sicau.edu.cn; PAN Deng-ke, E-mail: pandengke2002@163.com * These authors contributed equally to this study.
Supported by:
This work was supported by grants from the National Key R&D Program of China (2020YFA0509500), the National Natural Science Foundation of China (U19A2036, 32102512, 31872335, and 31802044), the National Special Foundation for Transgenic Species of China (2014ZX0800605B), the Sichuan Science and Technology Program, China (2021YFYZ0009 and 2021YFYZ0030).

摘要/Abstract

摘要：

本研究制备了在骨骼肌组织中特异表达人FST基因的转基因（TG）猪，并进行了表型鉴定。相较于野生型（WT）猪，TG猪骨骼肌重量显著增加(P<0.05)，脂肪沉积显著减少(P<0.05)，代谢状态显著改善(P<0.05)。根据表型变化，利用RNA-seq技术对WT猪和TG猪的骨骼肌（酵解型：背最长肌，氧化型：腰大肌）、白色脂肪（皮下脂肪：背部皮下脂肪，内脏脂肪：腹膜后脂肪）和肝脏共6个组织进行了转录组比较分析。结果表明，PIK3-AKT信号通路、钙离子信号通路及氨基酸代谢通路在FST诱导的骨骼肌肥大中具有重要作用；MYH7基因（决定I型肌纤维）表达量的相对比例在TG猪腰大肌中显著减低，氧化磷酸化和脂肪酸代谢等相关信号通路也在TG猪腰大肌中显著下调；相较于WT猪，TG猪脂肪中的AMPK信号通路、脂代谢相关通路发生显著变化，脂质合成、脂质分解及脂质储存相关基因表达量在皮下脂肪中显著降低，脂质分解相关基因表达量在腹膜后脂肪组织中显著升高。肝脏组织中，TGF-β信号通路相关基因在TG猪中显著下调。这些结果将有助于理解卵泡抑素引起猪表型变化的分子机制，为该候选靶点进一步在分子育种中的应用提供了基础数据。

Abstract:

Follistatin (FST) is an important regulator of skeletal muscle growth and adipose deposition through its ability to bind to several members of the transforming growth factor-β (TGF-β) superfamily, and thus may be a good candidate for future animal breeding programs. However, the molecular mechanisms underlying the phenotypic changes have yet to be clarified in pig. We generated transgenic (TG) pigs that express human FST specifically in skeletal muscle tissues and characterized the phenotypic changes compared with the same tissues in wild-type pigs. The TG pigs showed increased skeletal muscle growth, decreased adipose deposition, and improved metabolism status (P<0.05). Transcriptome analysis detected important roles of the PIK3–AKT signaling pathway, calcium-mediated signaling pathway, and amino acid metabolism pathway in FST-induced skeletal muscle hypertrophy, and depot-specific oxidative metabolism changes in psoas major muscle. Furthermore, the lipid metabolism-related process was changed in adipose tissue in the TG pigs. Gene set enrichment analysis revealed that genes related to lipid synthesis, lipid catabolism, and lipid storage were down-regulated (P<0.01) in the TG pigs for subcutaneous fat, whereas genes related to lipid catabolism were significantly up-regulated (P<0.05) in the TG pigs for retroperitoneal fat compared with their expression levels in wild-type pigs. In liver, genes related to the TGF-β signaling pathway were over-represented in the TG pigs, which is consistent with the inhibitory role of FST in regulating TGF-β signaling. Together, these results provide new insights into the molecular mechanisms underlying the phenotypic changes in pig.

Key words: follistatin , transgenic pig , transcriptome , skeletal muscle , liver , adipose

. JIA-2021-1043 转录组分析解析转人FST基因猪表型变化的分子机制[J]. Journal of Integrative Agriculture, 2022, 21(9): 2675-2690.

LONG Ke-ren, LI Xiao-kai, ZHANG Ruo-wei, GU Yi-ren, DU Min-jie, XING Xiang-yang, DU Jia-xiang, MAI Miao-miao, WANG Jing, JIN Long, TANG Qian-zi, HU Si-lu, MA Ji-deng, WANG Xun, PAN Deng-ke, LI Ming-zhou. Transcriptomic analysis elucidates the enhanced skeletal muscle mass, reduced fat accumulation, and metabolically benign liver in human follistatin-344 transgenic pigs[J]. Journal of Integrative Agriculture, 2022, 21(9): 2675-2690.

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JIA-2021-1043 转录组分析解析转人FST基因猪表型变化的分子机制

Transcriptomic analysis elucidates the enhanced skeletal muscle mass, reduced fat accumulation, and metabolically benign liver in human follistatin-344 transgenic pigs

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