IGF1-PI3K-Akt信号通路相关基因在黄羽肉鸡肌肉和肝脏中的表达

doi:10.3864/j.issn.0578-1752.2021.09.018

中国农业科学 ›› 2021, Vol. 54 ›› Issue (9): 2027-2038.doi: 10.3864/j.issn.0578-1752.2021.09.018

• 畜牧·兽医·资源昆虫 • 上一篇

IGF1-PI3K-Akt信号通路相关基因在黄羽肉鸡肌肉和肝脏中的表达

束婧婷¹(),姬改革¹,单艳菊¹,章明¹,巨晓军¹,刘一帆¹,屠云洁¹,盛中伟¹,唐燕飞²,蒋华莲²,邹剑敏¹()

¹江苏省家禽科学研究所/江苏省家禽遗传育种重点实验室,江苏扬州 225125
²广西富凤农牧集团有限公司,南宁 530024

收稿日期:2020-07-28 接受日期:2020-10-16 出版日期:2021-05-01 发布日期:2021-05-10
通讯作者: 邹剑敏
作者简介:束婧婷,E-mail：shujingting@163.com。
基金资助:
江苏省农业科技自主创新资金项目(CX203003);现代农业产业技术体系专项资金(CARS-41);江苏现代农业(肉鸡)产业技术体系集成创新中心(JATS[2019]379);江苏省农业重大新品种创制项目(PZCZ201728);江苏省家禽遗传育种重点实验室自主研究课题(JQLAB-ZZ-202001)

Expression Analysis of IGF1-PI3K-Akt-Dependent Pathway Genes in Skeletal Muscle and Liver Tissue of Yellow Feather Broilers

SHU JingTing¹(),JI GaiGe¹,SHAN YanJu¹,ZHANG Ming¹,JU XiaoJun¹,LIU YiFan¹,TU YunJie¹,SHENG ZhongWei¹,TANG YanFei²,JIANG HuaLian²,ZOU JianMin¹()

¹Jiangsu Institute of Poultry Science/Key laboratory for Poultry Genetics and Breeding of Jiangsu Province, Yangzhou 225125, Jiangsu
²Guangxi Fufeng Farm Group Co., Ltd., Nanning 530024

Received:2020-07-28 Accepted:2020-10-16 Online:2021-05-01 Published:2021-05-10
Contact: JianMin ZOU

摘要/Abstract

摘要：

【目的】IGF1-PI3K-Akt信号通路是调控生长发育的关键通路,选择慢速型的广西麻鸡和中速型的花山麻鸡作为研究对象,探究IGF1-PI3K-AKT信号通路相关基因胰岛素样生长因子1( Insulin-like growth factors, IGF1)、胰岛素样生长因子1受体( Insulin- like growth factor receptor, IGF1R)和胰岛素受体底物1 (Insulin receptor substrate1, IRS1)在骨骼肌和肝脏中的发育性表达规律,为阐明黄羽肉鸡生长发育调控机理提供参考。【方法】采用SPSS20.0比较广西麻鸡和花山麻鸡在胚胎期至出生后早期的生长发育差异,采用实时荧光定量PCR方法检测上述3个基因在两个品种鸡胚胎期（9、12、16胚龄）和出生后（0、7、21、35、49和63日龄）胸肌、腿肌和肝脏中的表达差异,并将其与体重和组织重进行相关性分析。【结果】鸡生长发育早期体重、骨骼肌重和肝脏重变化呈现显著的品种和时间特异性,经过选育的花山麻鸡体重和组织重的增长在胚胎发育后期已经远远超过广西麻鸡。IGF1、IGF1R和IRS1基因表达存在显著的品种、组织和发育时段特异性,总体上,在胚胎期肌肉中的表达量高于肝脏组织,出生后则为肝脏中的表达量高于肌肉组织,广西麻鸡肌肉中的表达量高于花山麻鸡,而花山麻鸡肝脏中的表达量要高于广西麻鸡。IGF1基因在两个品种胸腿肌中均在9胚龄即可检测到表达,腿肌中表达量均是9胚龄时最高,除与12胚龄差异不显著外,与其他各阶段差异显著,表达量最低点出现在出雏0日龄时;胸肌中则是12胚龄时表达量最高,63日龄时最低;肝脏中则出现品种差异,广西麻鸡胚胎期未检测到表达,从出雏0日龄开始表达,表达高峰出现在21日龄时,而在花山麻鸡中虽然胚胎期检测到表达但表达量极低,49日龄时达到表达高峰。两品种鸡肌肉和肝脏各个阶段均检测到IGF1R mRNA的表达,在胸肌和腿肌中的表达量均表现为9胚龄最高,广西麻鸡9胚龄与其他各阶段差异显著,花山麻鸡9胚龄除与12胚龄差异不显著外,与其他各阶段均差异显著;肝脏中,广西麻鸡表达高峰出现在21日龄,且与9、12、16胚龄和0日龄差异显著,花山麻鸡则是63日龄时最高,与其他各阶段差异显著。两品种鸡肌肉和肝脏各个阶段均能检测到IRS1 mRNA的表达,在两品种肌肉中的表达量均表现为9胚龄或者12胚龄最高,与其他各阶段差异显著,随后下降,并在出雏后维持在较低水平;肝脏中IRS1的表达模式与IGF1R一致。IGF1、IGF1R和IRS1基因的表达两两之间均表现出显著或极显著的正相关关系,同时,肌肉中3个基因的表达与体重、胸（腿）肌重存在显著的负相关,而肝脏中3个基因的表达与体重、胸（腿）肌重、肝脏重均存在显著的正相关。【结论】IGF1-PI3K-AKT信号通路相关基因之间存在一致性趋势,品种间和组织间表达量的差异可能正是不同类型黄羽肉鸡生长发育差异的主要原因之一。

关键词: 黄羽肉鸡, 基因表达, IGF1-PI3K-Akt信号通路, 肌肉, 肝脏

Abstract:

【Objective】IGF1-PI3K-Akt signal pathway have been implicated in the regulation of growth and development of chicken. In the present study, the expression of IGF, IGF1R and IRS1genes of IGF1-PI3K-Akt signal pathway was first studied in skeletal muscle and liver tissues of Guangxi partridge chickens with slow growth rate and Huashan partridge chickens with moderate growth rate, which would provide a reference for clarifying the regulation mechanism of growth and development of yellow feather broilers. 【Method】SPSS20.0 software was used to compare the differences of growth traits between Guangxi partridge chickens and Huashan partridge chickens. Expression of IGF1, IGF1R and IRS1 was quantified by RT-PCR in the breast muscle (BM), leg muscle (LM) and liver on days 9, 12, and 16 of embryonic development, as well as at 0, 7, 21, 35, 49, and 63 days post-hatching (PH) in the two chicken breeds, and the correlations between the gene expression level and body weight as well as tissue weight were also analyzed by SPSS20.0 software. 【Result】The body weight, skeletal muscle and liver weight of chicken in the early growth development showed significant breed- and age-specificity. The growth of Huashan partridge chickens had far exceeded that of Guangxi partridge chickens from the late embryonic development. The expression patterns of IGF1, IGF1R and IRS1 showed significant differences in breed-, tissue- and age-specific fashion. Overall, the expression levels of the three genes in skeletal muscles of embryonic development were much higher than those in liver; in contrast, the expression levels in liver were higher than those in skeletal muscle at post-hatching development. As for breed, expression levels of the studied genes in skeletal muscle of Guangxi partridge chickens were higher than those in Huashan partridge chickens; in contrast, liver expression levels were higher in Huashan partridge chickens. IGF1 mRNA could be detected as early as on E9 d in the skeletal muscles of both chicken breeds, and the highest level was appeared at this stage in LM, significantly higher than those on the other studied development stages expect on E12 d, and the lowest level was appeared on 0 d at hatching; while in BM, the highest level was appeared on E12 d and lowest level was appeared on 63 d; however, the level of IGF1 mRNA differed between breeds in liver. For Guangxi partridge chickens, the highest level was observed on 21 d and there was no expression during embryonic development, whereas the highest level was found on 49 d and very low level during embryonic development in Huashan partridge chickens. IGF1R mRNA could be detected at the whole studied stages in skeletal muscles and liver of both chicken breeds, and the highest level was appeared on E9 d in both BM and LM, significantly higher than those on the other studied development stages in Guangxi partridge chickens, and significantly higher than those on the other studied development stages expect on E12 d in Huashan partridge chickens. In liver, the highest expression level of IGF1R was detected on 21 d in Guangxi partridge chickens, significantly higher than those on E9 d, E12 d, E16 d and 0 d; while the highest level was found on 63 d and significantly higher than those on the other studied development stages. IRS1 mRNA could also be detected at the whole studied stages in skeletal muscles and liver of both chicken breeds. In skeletal muscles, expression of IRS1 were higher on E9 d or E12 d than any other age, and then decreased with age and kept relative low level during PH development in both breeds. The liver IRS1 expression pattern was consistent with IGF1R in both breeds. Significant positive relationships were observed for the expression of studied genes in BM, LM and liver tissues of both chicken breeds. Meanwhile, the skeletal muscle expression of these three genes were all showed significant negative relationships with body weight and breast (leg) muscle weight, whereas the liver expression of these three genes were all showed significant positive relationships with body weight, breast (leg) muscle weight and liver weight. 【Conclusion】These results implied that the expression of selected genes that comprise the IGF1-PI3K-Akt pathway existed identical trends, and differential expression between breeds and tissues might be one of the main reasons for the different growth rate of the different types of yellow feather broiler.

Key words: yellow feather broiler, gene expression, IGF1-PI3K-Akt pathway, skeletal muscle, liver

束婧婷,姬改革,单艳菊,章明,巨晓军,刘一帆,屠云洁,盛中伟,唐燕飞,蒋华莲,邹剑敏. IGF1-PI3K-Akt信号通路相关基因在黄羽肉鸡肌肉和肝脏中的表达[J]. 中国农业科学, 2021, 54(9): 2027-2038.

SHU JingTing,JI GaiGe,SHAN YanJu,ZHANG Ming,JU XiaoJun,LIU YiFan,TU YunJie,SHENG ZhongWei,TANG YanFei,JIANG HuaLian,ZOU JianMin. Expression Analysis of IGF1-PI3K-Akt-Dependent Pathway Genes in Skeletal Muscle and Liver Tissue of Yellow Feather Broilers[J]. Scientia Agricultura Sinica, 2021, 54(9): 2027-2038.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2021.09.018

https://www.chinaagrisci.com/CN/Y2021/V54/I9/2027

图/表 5

表1

图1

表2

广西麻鸡和花山麻鸡不同发育时期骨骼肌和肝脏中IGF1、IGF1R和IRS1基因的相对表达量"

基因Gene	组织Tissue	品种 Breed	9 ed	12 ed	16 ed	0 d	7 d	21 d	35 d	49 d	63 d
IGF1	胸肌 Breast	GXMJ	9.65±1.06 a	9.78±0.37a	5.18±1.07b	1.51±0.33 c	0.27±0.08d	0.19±0.05d	0.14±0.03d	0.06±0.005e	0.05±0.005e
	胸肌 Breast	HSMJ	1.12±0.18b**	1.92±0.26a**	0.98±0.26b**	0.42±0.07c*	0.24±0.08c	0.18±0.05c	0.17±0.03c	0.13±0.04c	0.05±0.004d
	腿肌 Leg	GXMJ	54.92±2.51a	54.77±5.06a	33.20±3.39 b	1.32±0.39c	2.08±0.37cd	2.86±0.44d	2.51±0.48cd	3.42±0.85d	1.58±0.16c
	腿肌 Leg	HSMJ	10.44±1.5a**	10.31±0.2a**	1.77±0.20b**	0.26±0.65c*	2.74±0.57d	1.53±0.26b	3.79±0.86d	2.25±0.42b	1.06±0.18b
	肝脏 Liver	GXMJ	0	0	0	1.23±0.23c	4.02±1.07c	57.62±6.46a	47.84±5.54a	52.47±7.05a	29.81±3.59b
	肝脏 Liver	HSMJ	0.04±0.01e*	0.22±0.02d*	0.31±0.04d	3.02±0.52c*	5.11±1.47c	66.06±6.99b*	74.44±9.4ab*	80.87±12.0a*	79.71±9.1a**
IGF 1R	胸肌 Breast	GXMJ	3.50±0.23 a	1.64±0.15b	2.63±0.33b	1.32±0.33 c	0.06±0.01d	0.08±0.01d	0.10±0.02d	0.03±0.00d	0.04±0.00d
	胸肌 Breast	HSMJ	0.45±0.07a**	0.24±0.04b**	0.25±0.06b**	0.17±0.07bc*	0.02±0.00c	0.02±0.00c	0.05±0.00c	0.03±0.00c	0.01±0.00c
	腿肌 Leg	GXMJ	6.14±0.86a	4.21±0.56b	2.31±0.37c	1.18±0.28d	0.46±0.08d	0.37±0.05d	0.49±0.05d	0.30±0.04d	0.21±0.06d
	腿肌 Leg	HSMJ	1.24±0.23a**	1.21±0.27a*	0.18±0.05b**	0.10±0.01b**	0.17±0.02b	0.12±0.02b	0.29±0.06b	0.17±0.02b	0.17±0.01b
	肝脏 Liver	GXMJ	0.15±0.02c	0.16±0.02 c	0.37±0.03c	0.96±0.06b	1.62±0.19ab	1.96±0.27a	1.48±0.16ab	1.54±0.19ab	1.35±0.28ab
	肝脏 Liver	HSMJ	0.21±0.05d	0.36±0.04d	0.50±0.03d	1.21±0.18c	1.77±0.23bc	1.78±0.30bc	1.88±0.24bc	2.60±0.35b*	5.74±0.91a*
IRS1	胸肌 Breast	GXMJ	2.07±0.09a	1.69±0.12a	1.51±0.16a	1.50±0.13a	0.17±0.02b	0.30±0.06b	0.37±0.03b	0.22±0.03b	0.21±0.02b
	胸肌 Breast	HSMJ	0.56±0.05a**	0.39±0.03a*	0.31±0.02a*	0.55±0.06a*	0.09±0.01c	0.16±0.01b	0.21±0.03b	0.13±0.04b	0.02±0.00c*
	腿肌 Leg	GXMJ	5.84±0.59a	4.32±0.49b	1.93±0.11c	1.51±0.04cd	0.79±0.06cd	1.07±0.13cd	0.90±0.10cd	0.80±0.06cd	0.61±0.05d
	腿肌 Leg	HSMJ	2.08±0.26a**	2.28±0.54a*	0.63±0.10b*	0.73±0.04b*	0.26±0.04b*	0.40±0.06b*	0.60±0.09b	0.48±0.07b	0.38±0.04b
	肝脏 Liver	GXMJ	0.09±0.00c	0.22±0.04c	0.10±0.01c	0.92±0.20b	1.80±0.27ab	3.08±0.46a	2.84±0.55a	3.02±0.74a	2.63±0.22a
	肝脏 Liver	HSMJ	0.08±0.00f	0.11±0.01f	0.08±0.01f	0.76±0.13e	1.52±0.11e	2.58±0.15d	3.53±0.53c	6.29±0.73b*	16.05±2.01a**

表2

表3

表4

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参数 Parameters		广西麻鸡Guangxi Partridge		花山麻鸡Huashan Partridge
参数 Parameters		IGF1R	IRS1	IGF1R	IRS1
胸肌 Breast muscle	IGF1	0.851**	0.641**	0.418**	0.457**
胸肌 Breast muscle	IRS1	0.806**		0.648**
腿肌 Leg muscle	IGF1	0.884**	0.873**	0.719**	0.670**
腿肌 Leg muscle	IRS1	0.854**		0.884**
肝脏 Liver	IGF1	0.494**	0.682**	0.489**	0.389**
肝脏 Liver	IRS1	0.759**		0.911**

组织 Tissue	参数 Parameters	广西麻鸡Guangxi Partridge			花山麻鸡Huashan Partridge
组织 Tissue	参数 Parameters	IGF1	IGF1R	IRS1	IGF1	IGF1R	IRS1
胸肌 Breast muscle	体重Body weight	-0.453**	-0.537**	-0.467**	-0.519*	-0.430**	-0.594**
胸肌 Breast muscle	胸肌重Breast weight	-0.405**	-0.483**	-0.424**	-0.441**	-0.361**	-0.542**
腿肌 Leg muscle	体重Body weight	-0.413**	-0.471**	-0.451**	-0.389**	-0.332**	-0.370**
腿肌 Leg muscle	腿肌重Leg weight	-0.377**	-0.435**	-0.417**	-0.362**	-0.284*	-0.321**
肝脏 Liver	体重Body weight	0.369*	0.965**	0.817**	0.693**	0.781**	0.817**
	肝脏重Liver weight	0.458**	0.409**	0.851**	0.601**	0.794**	0.851**
	胸肌重Breast weight	0.352**	0.453**	0.783**	0.563**	0.696**	0.752**
	腿肌重Leg weight	0.365**	0.483**	0.812**	0.598**	0.723**	0.795**

IGF1-PI3K-Akt信号通路相关基因在黄羽肉鸡肌肉和肝脏中的表达

Expression Analysis of IGF1-PI3K-Akt-Dependent Pathway Genes in Skeletal Muscle and Liver Tissue of Yellow Feather Broilers

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基因名称 Gene name	引物序列 Primer sequence	PCR条件 PCR conditions (℃)
IGF1	5′ primer: AGCTGGTTGATGCTCTTCAGTTCGT	60
IGF1	3′ primer: TCCTCAGGTCACAACTCTGGAAGC	60
IGF1R	5′ primer: TTCAGGAACCAAAGGGCGA	55
IGF1R	3′ primer: TGTAATCTGGAGGGCGATACC	55
IRS1	5′ primer: CTACCGCCTGTGCCTGACTAAC	60
IRS1	3′ primer: CGTCCCACCTCGATGAAGAAG	60
HSP70	5′ primer: TCTGCTCCTGTTGGATGTC	60
HSP70	3′ primer: TGGGAATGGTGGTGTTACG	60

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