A Characterization and Functional Analysis of Peroxisome Proliferator-Activated Receptor Gamma Splicing Variants in the Buffalo Mammary Gland
Author:
Wang Shuwan1, Ren Honghe1, Qin Chaobin1, Su Jie1, Song Xinhui1, Li Ruijia1, Cui Kuiqing2, Liu Yang3, Shi Deshun1ORCID, Liu Qingyou2, Li Zhipeng1ORCID
Affiliation:
1. Guangxi Key Laboratory of Animal Reproduction, Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China 2. Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan 528225, China 3. Guangxi Zhuang Autonomous Region Center for Analysis and Test Research, Nanning 530022, China
Abstract
Peroxisome proliferator-activated receptor γ (PPARG) has various splicing variants and plays essential roles in the regulation of adipocyte differentiation and lipogenesis. However, little is known about the expression pattern and effect of the PPARG on milk fat synthesis in the buffalo mammary gland. In this study, we found that only PPARG-X17 and PPARG-X21 of the splicing variant were expressed in the buffalo mammary gland. Amino acid sequence characterization showed that the proteins encoded by PPARG-X17 and PPARG-X21 are endonuclear non-secreted hydrophilic proteins. Protein domain prediction found that only the PPARG-X21-encoded protein had PPAR ligand-binding domains (NR_LBD_PPAR), which may lead to functional differences between the two splices. RNA interference (RNAi) and the overexpression of PPARG-X17 and PPARG-X21 in buffalo mammary epithelial cells (BMECs) were performed. Results showed that the expression of fatty acid synthesis-related genes (ACACA, CD36, ACSL1, GPAT, AGPAT6, DGAT1) was significantly modified (p < 0.05) by the RNAi and overexpression of PPARG-X17 and PPARG-X21. All kinds of FAs detected in this study were significantly decreased (p < 0.05) after RNAi of PPARG-X17 or PPARG-X21. Overexpression of PPARG-X17 or PPARG-X21 significantly decreased (p < 0.05) the SFA content, while significantly increased (p < 0.05) the UFA, especially the MUFA in the BMECs. In conclusion, there are two PPARG splicing variants expressed in the BMECs that can regulate FA synthesis by altering the expression of diverse fatty acid synthesis-related genes. This study revealed the expression characteristics and functions of the PPARG gene in buffalo mammary glands and provided a reference for further understanding of fat synthesis in buffalo milk.
Funder
Guangxi Natural Science Foundation Guangxi Science and Technology Major Project National Natural Science Foundation of China Guangdong Basic and Applied Research Foundation Guangxi Bagui Scholar Program
Reference32 articles.
1. Chen, J., and Liu, H. (2020). Nutritional indices for assessing fatty acids: A mini-review. Int. J. Mol. Sci., 21. 2. Fatty acids-from energy substrates to key regulators of cell survival, proliferation and effector function;Cucchi;Cell Stress,2020 3. Brodziak, A., Wajs, J., Zuba-Ciszewska, M., Król, J., Stobiecka, M., and Jańczuk, A. (2021). Organic versus conventional raw cow milk as material for processing. Animals, 11. 4. Fatty acid profiles of milk from Holstein cows, Jersey cows, buffalos, yaks, humans, goats, camels, and donkeys based on gas chromatography–mass spectrometry;Wang;J. Dairy Sci.,2022 5. Li, Z., Lu, S., Cui, K., Shafique, L., Rehman, S.U., Luo, C., Wang, Z., Ruan, J., Qian, Q., and Liu, Q. (2020). Fatty acid biosynthesis and transcriptional regulation of Stearoyl-CoA Desaturase 1 (SCD1) in buffalo milk. BMC Genet., 21.
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