Expression Analysis of TNNI1 and TNNI3 in Striated Muscle of Maiwa Yak

doi:10.7668/hbnxb.20192913

Abstract

Abstract:

In order to explore the expression patterns of TNNI1 and TNNI3 in the striated muscle of Maiwa yak,this study selectd the gluteal muscle and myocardial tissue of Maiwa yak of different ages,morphological changes observed.The expression trend of TNNI1 gene in gluteal muscle tissue of Maiwa yak at different ages and its association with muscle fiber traits were carried out,the mRNA,protein differential expression and protein localization of TNNI1 and TNNI3 in myocardium were analyzed.The results showed that the muscle fiber diameter and density had certain characteristic changes with the increase of yak age.The muscle fiber diameter increased significantly with the increase of age,while the muscle fiber density had the opposite change.The mRNA expression of TNNI1 gene in gluteal muscle tissue of adult yaks was significantly higher than calves and weaned calves. TNNI1 mRNA expression in gluteus muscle was significantly positively correlated with muscle fiber diameter,and significantly negatively correlated with muscle fiber density.The mRNA expression level of TNNI1 in myocardium decreased with the increase of yak age,while the expression level of TNNI3 showed a trend of first increased and then decreased,and the mRNA expression level of TNNI1 was the highest in calf stage,while TNNI3 showed an upward trend with the increase of age.The expression levels of TNNI1 and TNNI3 proteins in myocardium decreased with the increase of yak age.The expression levels of TNNI1 and TNNI3 proteins in myocardium decreased with the increase of yak age.The TNNI3 mRNA expression decreased with the increase of yak age,which was consistent with the trend of protein expression.It provided basic data for exploring the regulation mechanism of TNNI1 and TNNI3 on yak muscle growth and development.TNNI1 and TNNI3 have significant regulatory effects on the growth and development of the muscle tissue of Maiwa yak at different ages,and the expression levels in the muscle tissue of yak at different ages are different.

Key words: Yak, TNNI1, TNNI3, Striated muscle, Western Blot, Immunohistochemistry

XIANG Ya, ZHONG Jincheng, WU Zhijuan, CHAI Zhixin. Expression Analysis of TNNI1 and TNNI3 in Striated Muscle of Maiwa Yak[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(6): 210-218. doi: 10.7668/hbnxb.20192913.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20192913

http://www.hbnxb.net/EN/Y2022/V37/I6/210

Figures/Tables 14

Tab.1 Primers for PCR

基因名称 Genes	引物序列(5'—3') Oligo sequence(5'—3')	片段长度/bp Fragment length	退火温度/℃ Annealing temperature
TNNI1	F:CTCCAAGCACAAGGTGTCTATG	158	56
	R:TGGCGGCATCAAACATCT
TNNI3	F:CTCGTGTGGACAAGGTGGA	137	58
	R:GCCCCGAAGGTCAAAGAT
GAPDH	F:CCACGAGAAGTATAACAACACC	120	55
	R:GTCATAAGTCCCTCCACGAT

Fig.1 Expression level of TNNI1 in gluteal muscle tissues of Maiwa yak Different lowercase letters indicate significant difference (P<0.05).The same as Fig.2,5,6,9—11.

Fig.2 Expression of TNNI1 and TNNI3 in Maiwa yak myocardium Different capital letters indicate significant difference between two genes(P<0.05).

Fig.3 Protein absorbance standard curve

Fig.4 SDS-PAGE electrophoresis results of myocardial protein 1—3.Adult yak;4—6.Weaned calf;7—9.Calf.

Fig.5 Western Blot analysis of TNNI1

Fig.6 Western Blot analysis of TNNI3

Fig.7 Gluteal muscle fibers slices of Maiwa yak at different ages(10×10) A.Calf;B.Weaned calf;C.Adult yak.The same as Fig.8—11.

Tab.2 Gluteal muscle fiber traits comparison of Maiwa yak at different ages

年龄段 Age	肌纤维直径/μm Muscle fiber diameter	肌纤维密度 /(N/mm²) Muscle fiber density
犊牛Calf	65.26±9.79b	4 437.45±280.29a
断奶犊牛Weaned calf	69.79±6.00b	4 207.25±118.11a
成年牛Adult yak	142.31±13.09a	2 880.91±69.83b

Fig.8 Myocardial muscle fibers slices of Maiwa yak at different ages

Tab.3 Correlations between TNNI1 mRNA expression and muscle fiber traits

mRNA表达 mRNA expression	项目 Items	相关系数 Correlation coefficient	P值 P value
TNNI1	肌纤维直径	0.95	0.004
	肌纤维密度	-0.90	0.016

Fig.9 Protein expression characteristics of TNNI1 in gluteal muscle(10×10)

Fig.10 Protein expression characteristics of TNNI1 in myocardial(10×10)

Fig.11 Protein expression characteristics of TNNI3 in myocardial(10×10)

References 35

[1]	Robaszkiewicz K, Ostrowska Z, Cyranka-Czaja A, Moraczewska J. Impaired tropomyosin-troponin interactions reduce activation of the actin thin filament[J]. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics, 2015, 1854(5): 381-390.doi:10.1016/j.bbapap.2015.01.004. doi: 10.1016/j.bbapap.2015.01.004 pmid: 25603119
[2]	Sheng J J, Jin J P. TNNI1,TNNI2 and TNNI3: Evolution,regulation,and protein structure-function relationships[J]. Gene, 2016, 576(1): 385-394.doi:10.1016/j.gene.2015.10.052. doi: 10.1016/j.gene.2015.10.052 URL
[3]	Perry S V. Troponin T: Genetics,properties and function[J]. Journal of Muscle Research and Cell Motility, 1998, 19(6): 575-602.doi:10.1023/a:1005397501968. doi: 10.1023/a:1005397501968 pmid: 9742444
[4]	吴婷婷. 天府肉羊TNNI1和TNNI3基因的克隆、表达及其与肌肉组织学性状相关性研究[D]. 雅安: 四川农业大学, 2013.
	Wu T T. Cloning and expression of Tianfu goat TNNI1 and TNNI3 gene and their relationship with muscle tissue traits[D]. Yaan: Sichuan Agricultural University, 2013.
[5]	姬改革, 束婧婷, 单艳菊, 章明, 屠云洁, 巨晓军, 刘一帆. 高邮鸭TNNI1基因克隆及组织表达分析[J]. 农业生物技术学报, 2018, 26(6): 970-977.doi:10.3969/j.issn.1674-7968.2018.06.007. doi: 10.3969/j.issn.1674-7968.2018.06.007
	Ji G G, Shu J T, Shan Y J, Zhang M, Tu Y J, Ju X J, Liu Y F. Molecular cloning and tissue expression analysis of TNNI1 gene in Gaoyou duck(Anas platyrhynchos)[J]. Journal of Agricultural Biotechnology, 2018, 26(6): 970-977.
[6]	杨华. 猪肌钙蛋白Ⅰ基因家族的基因结构、表达谱和多态性分析[D]. 武汉: 华中农业大学, 2009.
	Yang H. Porcine troponin Ⅰ gene family: Genomic structure,expression profiling and polymorphism analysis[D]. Wuhan: Huazhong Agricultural University, 2009.
[7]	Sasse S, Brand N J, Kyprianou P, Dhoot G K, Wade R, Arai M, Periasamy M, Yacoub M H, Barton P J. Troponin I gene expression during human cardiac development and in end-stage heart failure[J]. Circulation Research, 1993, 72(5): 932-938.doi:10.1161/01.res.72.5.932. doi: 10.1161/01.res.72.5.932 pmid: 8477526
[8]	Saggin L, Gorza L, Ausoni S, Schiaffino S. Troponin I switching in the developing heart[J]. Journal of Biological Chemistry, 1989, 264(27): 16299-16302.doi:10.1016/S0021-9258(18)71621-9. doi: 10.1016/S0021-9258(18)71621-9 pmid: 2777792
[9]	Murphy A M, Jones L, Sims H F, Strauss A W. Molecular cloning of rat cardiac troponin I and analysis of troponin I isoform expression in developing rat heart[J]. Biochemistry, 1991, 30(3): 707-712.doi:10.1021/bi00217a018. doi: 10.1021/bi00217a018 pmid: 1988058
[10]	Bhavsar P K, Brand N J, Yacoub M H, Barton P J R. Isolation and characterization of the human cardiac troponin I gene(TNNI3)[J]. Genomics, 1996, 35(1): 11-23.doi:10.1006/geno.1996.0317. doi: 10.1006/geno.1996.0317 pmid: 8661099
[11]	向娅, 柴志欣, 王吉坤, 信金伟, 王会, 王嘉博, 武志娟, 钟金城, 姬秋梅. 牦牛肌钙蛋白Ⅰ基因家族的克隆及组织表达分析[J]. 华北农学报, 2020, 35(2): 228-238.doi: 10.7668/hbnxb.20190667. doi: 10.7668/hbnxb.20190667
	Xiang Y, Chai Z X, Wang J K, Xin J W, Wang H, Wang J B, Wu Z J, Zhong J C, Ji Q M. Cloning and tissue expression of troponin Ⅰ gene family in yak[J]. Acta Agriculturae Boreali-Sinica, 2020, 35(2): 228-238.
[12]	张阳. 小尾寒羊TNNI2和TNNI3基因的克隆、结构分析及组织表达分析[D]. 泰安: 山东农业大学, 2016.
	Zhang Y. Molecular cloning,characterization and tissue expression analysis of the TNNI2 and TNNI3 from small-tailed Han sheep[D]. Taian: Shandong Agricultural University, 2016.
[13]	王飞龙. 骨提取蛋白修复大鼠颅骨缺损的实验研究[D]. 大连: 大连医科大学, 2021.
	Wang F L. Experimental study on repairing rat skull defect with bone protein extract[D]. Dalian: Dalian Medical University, 2021.
[14]	段小果. 大青山山羊肉肌纤维特性及品质的研究[D]. 呼和浩特: 内蒙古农业大学, 2017.
	Duan X G. Study on muscle fiber characteristics and meat quality of Daqingshan goat[D]. Hohhot: Inner Mongolia Agricultural University, 2017.
[15]	徐咏书. 髓系来源抑制性细胞通过Wnt/β-catenin信号通路促进肺腺癌进行干细胞样转变的分子机制研究[D]. 南昌: 南昌大学, 2020.
	Xu Y S. Myeloid-derived suppressor cell promotes the stem cell-like acquisition of lung adenocarcinoma through Wnt/β-catenin pathway[D]. Nanchang: Nanchang University, 2020.
[16]	Xu Z Y, Yang H, Li Y, Xiong Y Z, Zuo B. Temporal expression of TnI fast and slow isoforms in biceps femoris and masseter muscle during pig growth[J]. Animal, 2010, 4(9): 1541-1546.doi:10.1017/S1751731110000649. doi: 10.1017/S1751731110000649 pmid: 22444701
[17]	姬改革, 单艳菊, 章明, 屠云洁, 巨晓军, 张笛, 束婧婷. 鸭发育早期骨骼肌TNNI1基因mRNA表达及与肌纤维生长的相关性分析[J]. 安徽农业大学学报, 2017, 44(6): 992-995.doi:10.13610/j.cnki.1672-352x.20171214.015. doi: 10.13610/j.cnki.1672-352x.20171214.015
	Ji G G, Shan Y J, Zhang M, Tu Y J, Ju X J, Zhang D, Shu J T. Expression profiles of the TNNI1 gene in the skeletal muscle and its correlation with the muscle fiber growth in early developmental ducks[J]. Journal of Anhui Agricultural University, 2017, 44(6): 992-995.
[18]	Zheng Q, Zhang Y, Chen Y, Yang N, Wang X J, Zhu D H. Systematic identification of genes involved in divergent skeletal muscle growth rates of broiler and layer chickens[J]. BMC Genomics, 2009, 10: 87.doi:10.1186/1471-2164-10-87. doi: 10.1186/1471-2164-10-87 pmid: 19232135
[19]	Corin S J, Juhasz O, Zhu L, Conley P, Kedes L, Wade R. Structure and expression of the human slow twitch skeletal muscle troponin I gene[J]. Journal of Biological Chemistry, 1994, 269(14): 10651-10659.doi:10.1016/S0021-9258(17)34109-1. doi: 10.1016/S0021-9258(17)34109-1 pmid: 8144655
[20]	钟金城. 牦牛遗传与育种[M]. 成都: 四川科学技术出版社,1996.
	Zhong J C. Yak genetics and breeding[M]. Chengdu: Sichuan Science and Technology Press,1996.
[21]	Bhavsar P K, Dellow K A, Yacoub M H, Brand N J, Barton P J. Identification of Cis-acting DNA elements required for expression of the human cardiac troponin I gene promoter[J]. Journal of Molecular and Cellular Cardiology, 2000, 32(1): 95-108.doi:10.1006/jmcc.1999.1058. doi: 10.1006/jmcc.1999.1058 pmid: 10652194
[22]	Bennett Murphy L M, Thompson R J, Morris M A. Adherence behavior among adolescents with type I insulin-dependent diabetes mellitus: The role of cognitive appraisal processes[J]. Journal of Pediatric Psychology, 1997, 22(6): 811-825.doi:10.1093/jpepsy/22.6.811. doi: 10.1093/jpepsy/22.6.811 pmid: 9494319
[23]	肖成, 曹阳, 于永生, 张立春, 魏天, 高一, 金海国. TNNI基因对家畜肉质性状的影响进展[J]. 东北农业科学, 2020, 45(5):69-72.doi:10.16423/j.cnki.1003-8701.2020.05.018. doi: 10.16423/j.cnki.1003-8701.2020.05.018
	Xiao C, Cao Y, Yu Y S, Zhang L C, Wei T, Gao Y, Jin H G. Progress in the effect of TNNI gene on meat quality traits of livestock[J]. Journal of Northeast Agricultural Sciences, 2020, 45(5): 69-72.
[24]	刘伟娟. 几种肉羊肌肉组织学性状和理化性状的研究[D]. 保定: 河北农业大学, 2009.doi:10.7666/d.d144466. doi: 10.7666/d.d144466
	Liu W J. Study on the histological and physicochemical characteristics of muscle in mutton sheep[D]. Baoding: Hebei Agricultural University, 2009.
[25]	周金星, 毕亚玲, 高登慧. 肌肉组织学结构及其与肉品品质的关系[J]. 山地农业生物学报, 2004, 23(5): 438-441.doi:10.3969/j.issn.1008-0457.2004.05.013. doi: 10.3969/j.issn.1008-0457.2004.05.013
	Zhou J X, Bi Y L, Gao D H. A study on the structure of muscle histology and meat quality[J]. Journal of Mountain Agriculture and Biology, 2004, 23(5): 438-441.
[26]	毛国祥, 赵万里. 新太湖鹅、太湖鹅和隆昌鹅肌肉品质比较研究[J]. 动物科学与动物医学, 2000, 17(1): 16-19.
	Mao G X, Zhao W L. Comparison on muscle quality of Longchang,Taihu and new Taihu goose[J]. Animal Science and Veterinary Medicine, 2000, 17(1): 16-19.
[27]	McMeekan C P. Growth and development in the pig,with special reference to carcass quality characters: Ⅲ.Effect of the plane of nutrition on the form and composition of the bacon pig[J]. The Journal of Agricultural Science, 1940, 30(4): 511-569.doi:10.1017/s002185960004822x. doi: 10.1017/s002185960004822x URL
[28]	Carpenter F G. Impairment and restoration of rat urinary bladder responsiveness following distension[J]. The American Journal of Physiology, 1983, 244(1): R106-R113.doi:10.1152/ajpregu.1983.244.1.R106. doi: 10.1152/ajpregu.1983.244.1.R106
[29]	杨博辉, 姚军, 王敏强, 陆仲磷, 柏家林, 阎萍, 梁春年. 大通牦牛肌肉纤维组织学特性研究[J]. 中国草食动物, 2001, 21(5): 34-35.doi:10.3969/j.issn.2095-3887.2001.05.04. doi: 10.3969/j.issn.2095-3887.2001.05.04
	Yang B H, Yao J, Wang M Q, Lu Z L, Bai J L, Yan P, Liang C N. Study on histologic characteristics of muscle fiber in Datong yak[J]. China Herbivores, 2001(5):34-35.
[30]	孙竹珑, 钟光辉. 九龙牦牛肌肉组织学特性研究(初报)[J]. 西南民族学院学报(自然科学版), 1990, 16(3): 31-36.
	Sun Z L, Zhong G H. Study on the histological characters of muscle in Jiulong yak[J]. Journal of Southwest University for Nationalities (Natural Science Edition), 1990, 16(3): 31-36.
[31]	沈元新, 徐继初. 金华猪及其杂种肌肉组织学特性与肉质的关系[J]. 浙江农业大学学报, 1984, 10(3): 29-36.
	Shen Y X, Xu J C. The relationship between the muscle tissue characteristics of Jinhua swine and its hybrid and its meat quality[J]. Journal of Zhejiang Agricultural University, 1984, 10(3): 29-36.
[32]	孙艳. 小尾寒羊三个慢型骨骼肌肌钙蛋白的基因克隆、结构特征及组织表达分析[D]. 泰安: 山东农业大学, 2016.doi:10.7666/d.D833596. doi: 10.7666/d.D833596
	Sun Y. Molecular cloning,characterization and tissue expression analysis of three slow skeletal muscle troponin genes in small-tailed Han sheep[D]. Taian: Shandong Agricultural University, 2016.
[33]	张璐璐. Tnni3基因协同Tbx20基因对大鼠心肌细胞H9C2细胞凋亡的影响及机制研究[D]. 沈阳: 中国医科大学, 2019.
	Zhang L L. Biological effect and mechanism of synergistic interaction between Tnni3 gene and Tbx20 gene on rat H9C2 cardiomyocyte apoptosis[D]. Shenyang: China Medical University, 2019.
[34]	蔡陈. 斑马鱼tnni1b基因敲除导致心脏房室瓣膜发育异常的机制研究[D]. 武汉: 华中科技大学, 2018.
	Cai C. Research on the mechanism of the defects in atrioventricular valve development caused by the tnni1b knockout in zebrafish[D]. Wuhan: Huazhong University of Science and Technology, 2018.
[35]	桑彩君. 斑马鱼tnni1b缺失通过ERBB-ERK信号通路调控心脏瓣膜发育[D]. 武汉: 华中科技大学, 2019.
	Sang C J. The heart valve development defects caused by the tnni1b deletion in zebrafish via the ERBB-ERK signaling pathway[D]. Wuhan: Huazhong University of Science and Technology, 2019.

Please choose a citation manager

Content to export