G3BP1基因在鸡肌内前脂肪细胞增殖与分化中的作用及其分子标记鉴定

doi:10.11843/j.issn.0366-6964.2025.01.015

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (1): 159-167.doi: 10.11843/j.issn.0366-6964.2025.01.015

G3BP1基因在鸡肌内前脂肪细胞增殖与分化中的作用及其分子标记鉴定

李远方¹^,²(), 吴冉³^,⁴, 李帅浩³^,⁴, 魏千然¹^,², 王亚东¹^,², 王丹丹⁵, 李智¹^,²^,*(), 李国喜³^,⁴^,*(), 刘翘铭¹^,²^,*()

1. 哈尔滨工业大学医学与健康学院, 哈尔滨 150001
2. 哈尔滨工业大学郑州研究院, 郑州 450000
3. 神农种业实验室, 郑州 450002
4. 河南农业大学动物科技学院, 郑州 450046
5. 中国农业科学院北京畜牧兽医研究所, 北京 100193

收稿日期:2024-08-09 出版日期:2025-01-23 发布日期:2025-01-18
通讯作者: 李智,李国喜,刘翘铭 E-mail:yuanfangli1991@163.com;18530085133@163.com;liguoxi0914@126.com;cslqm@hit.edu.com
作者简介:李远方(1991-)，女，河南平顶山人，博士，主要从事畜禽资源分子评价的研究，E-mail: yuanfangli1991@163.com
基金资助:
鸡生长性状精准评价与高产优质核心种质创制(SN01-2022-05)；重要农业动植物优异性状共性调控元件挖掘与基因资源设计(2023YFF1001100)；中国博士后科学基金面上项目(GZC20233467)

The Role of G3BP1 in the Proliferation and Differentiation of Chicken Intramuscular Preadipocytes and Identification of Its Molecular Markers

LI Yuanfang¹^,²(), WU Ran³^,⁴, LI Shuaihao³^,⁴, WEI Qianran¹^,², WANG Yadong¹^,², WANG Dandan⁵, LI Zhi¹^,²^,*(), LI Guoxi³^,⁴^,*(), LIU Qiaoming¹^,²^,*()

1. School of Medicine and Health, Harbin Institute of Technology, Harbin 150001, China
2. Zhengzhou Research Institute, Harbin Institute of Technology, Zhengzhou 450000, China
3. The Shennong Laboratory, Zhengzhou 450002, China
4. College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China
5. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2024-08-09 Online:2025-01-23 Published:2025-01-18
Contact: LI Zhi, LI Guoxi, LIU Qiaoming E-mail:yuanfangli1991@163.com;18530085133@163.com;liguoxi0914@126.com;cslqm@hit.edu.com

摘要/Abstract

摘要：

旨在利用721只固始鸡F2代鸡群，鉴定影响肌内脂肪沉积的候选基因G3BP1的SNPs位点，用于肉质性状的分子标记开发，并利用固始鸡肌内前脂肪细胞深入探究其功能。本研究首先鉴定了G3BP1基因上的SNPs位点，发现3个突变位点处于强连锁不平衡状态，尤其是13588667G>A位点的不同基因型与固始鸡F₂资源群的皮脂、皮脂率和肌内脂肪等肉质性状显著相关(P < 0.05)。因此，G3BP1基因13588667G>A位点可以作为影响肌内脂肪沉积的一个重要的分子标记。此外，本研究利用qRT-PCR、CCK-8、流式细胞术、甘油三酯含量测定和油红O染色等方法，评价了G3BP1基因对固始鸡肌内前脂肪细胞增殖与分化的影响。结果表明，过表达G3BP1后，肌内前脂肪细胞中增殖标志基因CDK1、CCNB2、PCNA和CCND1表达水平均极显著上升(P < 0.001)，并显著促进了细胞周期进程。过表达G3BP1后也显著促进了肌内前脂肪细胞分化标志基因LPL的表达(P < 0.05)，极显著促进了CEBPA的表达(P < 0.001)，肌内前脂肪细胞内脂滴生成和甘油三酯含量也极显著增加(P < 0.01)。本研究明确了G3BP1在促进肌内前脂肪细胞增殖与脂肪细胞生成中的作用，并鉴定了影响肌内脂肪沉积的分子标记，为我国优质肉鸡培育提供重要的理论依据和应用价值。

关键词: 固始鸡, 肌内脂肪, G3BP1, 分子标记

Abstract:

The study aimed to identify SNP sites of the candidate gene G3BP1 that influence intramuscular fat deposition using a population of 721 Gushi F2 chickens, to develop molecular markers for meat quality traits, and to further investigate its function using intramuscular preadipocytes from Gushi chickens. First, the study identified SNPs in G3BP1 gene and found 3 mutation sites in strong linkage disequilibrium, especially the different genotypes of the 13588667G>A site were significantly associated with the meat quality traits such as sebum, sebum rate and intramuscular fat in the F2 resource group of Gushi chicken (P < 0.05). Therefore, the 13588667G>A site of the G3BP1 gene could be used as an important molecular marker affecting intramuscular fat deposition. Additionally, qRT-PCR, CCK-8, flow cytometry, triglyceride content determination and Oil Red O staining were used to evaluate the effect of G3BP1 gene on the proliferation and differentiation of intramuscular preadipocytes in Gushi chickens. The results showed that after overexpression of G3BP1, the expression levels of proliferation marker genes CDK1, CCNB2, PCNA and CCND1 in intramuscular preadipocytes were significantly increased (P < 0.001), and the cell cycle process was significantly promoted. G3BP1 overexpression also significantly upregulated the expression of the differentiation marker gene LPL (P < 0.05) and dramatically increased the expression of CEBPA (P < 0.001), with a significant increase in lipid droplet formation and triglyceride content in intramuscular preadipocytes (P < 0.01). This study clarifies the role of G3BP1 in promoting the proliferation and adipogenesis of intramuscular preadipocytes and identifies a molecular marker affecting intramuscular fat deposition, providing valuable theoretical and practical insights for breeding high-quality meat chickens in China.

Key words: Gushi chicken, intramuscular fat, G3BP1, molecular marker

中图分类号:

S831.2

李远方, 吴冉, 李帅浩, 魏千然, 王亚东, 王丹丹, 李智, 李国喜, 刘翘铭. G3BP1基因在鸡肌内前脂肪细胞增殖与分化中的作用及其分子标记鉴定[J]. 畜牧兽医学报, 2025, 56(1): 159-167.

LI Yuanfang, WU Ran, LI Shuaihao, WEI Qianran, WANG Yadong, WANG Dandan, LI Zhi, LI Guoxi, LIU Qiaoming. The Role of G3BP1 in the Proliferation and Differentiation of Chicken Intramuscular Preadipocytes and Identification of Its Molecular Markers[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 159-167.

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基因Gene	引物序列(5′→3′)Primer sequence	片段长度/bp Length	退火温度/℃ Tm
G3BP1	F: CTCCAAATCCTGGCTTCT R: GAGGGTCTGCTGTTCTTCAT	563	60
CCNB2	F: CCTCTTCCACTTCACTTCT R: CTTTGTACCCCACTTATCA	195	60
CCND1	F: CAGAAGTGCGAAGAGGAAGT R: CTGATGGAGTTGTCGGTGTA	188	60
PCNA	F: AGCACCAAATCAGGAAAAG R: GCACAGGAGATGACAACAG	177	60
CDK1	F: TAATAGATGACAAAGGGGT R: GAGTGGAATACAGAGCAGA	147	60
LPL	F: TCCGATCCCGAAGCTGAGA R: ACATTCCTGTCACCGTCCAC	167	60
FABP4	F: ATATGAAAGAGCTGGGTGTGGG R: CAGTGTGCCATTGTCTAGGGT	218	60
FABP5	F: TGGCCATCGACGCGTTTTTA R: TCTGGTTTCGCCATGCTTCC	121	60
FASN	F: ACAACAGCCAGCTTGGAATG R: CAGGGGCTTTACCACACCAT	152	60
PPARγ	F: GTGCAATCAAAATGGAGCC R: CTTACAACCTTCACATGCAT	170	60
CEBPA	F: TTCTACGAGGTCGATTCCCG R: AGCCTCTCTGTAGCCGTAG	96	60
GAPDH	F: GAACATCATCCCAGCGTCCA R: CGGCAGGTCAGGTCAACAAC	132	60

突变位点 SNP site	数量 Number	基因型频率 Genotype frequency			等位基因频率 Allele frequency		杂合度 Heterozygosity
突变位点 SNP site	数量 Number	II	ID	DD	I	D	杂合度 Heterozygosity
13588667G>A	721	0.681	0.288	0.030	0.836	0.164	0.274
13588768C>A	721	0.575	0.362	0.062	0.768	0.232	0.356
13588930C>T	721	0.241	0.447	0.312	0.438	0.562	0.492

相关指标 Relevant metrics	基因型 Genotype			P值 P-value
相关指标 Relevant metrics	GG	GA	AA	P值 P-value
皮脂/g Sebum	7.316±0.522^b	9.667±0.799^a	5.898±2.454^ab	0.033
皮脂率/% Sebum rate	0.801±0.055^b	1.038±0.085^a	0.649±0.264^b	0.045
肌间脂肪宽/cm Intermuscular fat width	0.729±0.012	0.737±0.019	0.720±0.058	0.929
皮下脂肪厚/cm Subcutaneous fat thickness	0.457±0.008	0.464±0.012	0.454±0.035	0.862
鲜样脂肪含量/% Fat content in fresh samples	0.719±0.013	0.705±0.019	0.711±0.060	0.833
绝干脂肪含量/% Fat content in absolute dry samples	2.747±0.047	2.705±0.072	2.746±0.222	0.886
肌内脂肪含量/% Intramuscular fat content	2.681±0.031^a	2.437±0.047^b	2.244±0.145^b	＜0.001

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G3BP1基因在鸡肌内前脂肪细胞增殖与分化中的作用及其分子标记鉴定

The Role of G3BP1 in the Proliferation and Differentiation of Chicken Intramuscular Preadipocytes and Identification of Its Molecular Markers

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