Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (9): 2027-2038.doi: 10.3864/j.issn.0578-1752.2021.09.018

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles    

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()   

  1. 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 E-mail:shujingting@163.com;jqszjm@163.com

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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

Table 1

Gene-specific primers for quantitative RT-PCR"

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

Fig. 1

The profiles of body weight, breast muscle weight, thigh muscle weight and liver weight during growth stages in different chicken breeds A: Body weight; B: Breast muscle weight; C: Leg muscle weight; D: Liver weight. ed indicates embryonic days, and d indicates age after hatching. Bar diagram values with the same letter are not significantly different between different age (P>0.05), and values with the different letters are significantly different between different ages (P<0.05). Superscript * indicates significantly different between two breeds in the same age at the level of 0.05, and superscript ** indicates significantly different between two breeds in the same age at the level of 0.01"

Table 2

The relative expression level of IGF1, IGF1R and IRS1 in skeletal muscles and liver of Huashan and Guangxi partridge chickens at different developmental stages"

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
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
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
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
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
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
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
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
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
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**

Table 3

Correlations betweenIGF1-PI3K-AKT signaling pathway genes in skeletal muscles and liver of the two chicken breeds"

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

Table 4

Correlations between the expression of IGF1-PI3K-AKT signaling pathway genes and growth traits in the two chicken breeds"

组织
Tissue		 参数
Parameters		 广西麻鸡Guangxi Partridge 花山麻鸡Huashan Partridge
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**
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