Overexpression of cholinergic receptor nicotinic gamma subunit inhibits proliferation and differentiation of bovine preadipocytes

Jiawei Du; Hui Zhao; Guibing Song; Yuan Pang; Lei Jiang; Linsen Zan; Hongbao Wang

doi:10.5713/ab.22.0144

Animal Bioscience (Feb 2023)

Overexpression of cholinergic receptor nicotinic gamma subunit inhibits proliferation and differentiation of bovine preadipocytes

Jiawei Du,
Hui Zhao,
Guibing Song,
Yuan Pang,
Lei Jiang,
Linsen Zan,
Hongbao Wang

Affiliations

Jiawei Du: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
Hui Zhao: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
Guibing Song: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
Yuan Pang: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
Lei Jiang: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
Linsen Zan: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China
Hongbao Wang: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China

DOI: https://doi.org/10.5713/ab.22.0144
Journal volume & issue: Vol. 36, no. 2
pp. 200 – 208

Abstract

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Objective Muscle acetylcholine receptors have five alpha subunits (α, β, δ, ɛ, or γ), and cholinergic receptor nicotinic gamma subunit (CHRNG) is the γ subunit. It may also play an essential role in biological processes, including cell differentiation, growth, and survival, while the role of CHRNG has not been studied in the literature. Therefore, the purpose of this study is to clarify the effect of CHRNG on the proliferation and differentiation of bovine preadipocytes. Methods We constructed a CHRNG overexpression adenovirus vector and successfully overexpressed it on bovine preadipocytes. The effects of CHRNG on bovine preadipocyte proliferation were detected by Edu assay, cell counting Kit-8 (CCK-8), real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), Western blot and other techniques. We also performed oil red O, RT-qPCR, Western blot to explore its effect on the differentiation of preadipocytes. Results The results of Edu proliferation experiments showed that the number of EDU-positive cells in the overexpression group was significantly less. CCK-8 experiments found that the optical density values of the cells in the overexpression group were lower than those of the control group, the mRNA levels of proliferating cell nuclear antigen (PCNA), cyclin A2 (CCNA2), cyclin B1 (CCNB1), cyclin D2 (CCND2) decreased significantly after CHRNG gene overexpression, the mRNA levels of cyclin dependent kinase inhibitor 1A (CDKN1A) increased significantly, and the protein levels of PCNA, CCNB1, CCND2 decreased significantly. Overexpression of CHRNG inhibited the differentiation of bovine preadipocytes. The results of oil red O and triglyceride determination showed that the size and speed of lipid droplets accumulation in the overexpression group were significantly lower. The mRNA and protein levels of peroxisome proliferator activated receptor gamma (PPARγ), CCAAT enhancer binding protein alpha (CEBPα), fatty acid binding protein 4 (FABP4), fatty acid synthase (FASN) decreased significantly. Conclusion Overexpression of CHRNG in bovine preadipocytes inhibits the proliferation and differentiation of bovine preadipocytes.

Published in Animal Bioscience

ISSN: 2765-0189 (Print); 2765-0235 (Online)
Publisher: Asian-Australasian Association of Animal Production Societies
Country of publisher: Korea, Republic of
LCC subjects: Science: Zoology
Website: https://www.animbiosci.org/index.php

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