Expression of calpastatin gene in Raini Cashmere goat using Real Time PCR

Document Type : Research Paper

Author

Department of Animal Science, Faculty of Agriculture, College of Agriculture, Shahid Bahonar University of Kerman

Abstract

Objective
Calpastatin has been found in skeletal muscle tissue of animals and could prevent unrestrained cellular growth by suppressing calpain activity. The involvement of calpains in apoptosis has been a subject of debate, although their involvement is limited to certain cell types and to specific stimuli. Various studies have shown that this gene is involved in growth performance and meat quality, therefore the aim of this study was to investigate Calpastatin gene expression in different tissues of Raini cashmere goat using Real Time PCR.
 
Materials and Methods
Totally 21 tissue samples including muscle, adipose, kidney, spleen, liver, lung and heart tissues were taken from 3 Raini cashmere goats. RNA was extracted and cDNA was synthesized. Real Time PCR was performed using SYBR Green method to study relative gene expression. GAPDH gene was used as housekeeping gene. Pfaffl method was used to analyze achieved data.
 
Results  
For calpastatin gene 89bp fragment and for GAPDH 101bp fragment were observed in all studied tissues. Real Time PCR results of this study showed that Calpastatin gene is expressed in all studied (muscle, adipose, kidney, spleen, liver, lung and heart) tissues and the highest level of expression was observed in heart, spleen and liver tissues and the lowest level was seen in adipose tissue.
 
Conclusions
Results showed that Calpastatin are widespread in different tissues of goat. This study would also lay a foundation for further Calpastatin research in Raini cashmere goat. It is suggested that this study be conducted with greater number of livestock, different sexes, different ages and different physiological stages in different breeds of goats in order to reach a comprehensive conclusion.

Keywords


منابع
بهرام پور وحید، محمدآبادی محمدرضا، میرزایی حمیدرضا و همکاران (1387) آنالیز ملکولی ژن کالپاستاتین در گله های گوسفند کرمانی. مجله علوم کشاورزی و منابع طبیعی 15(4)، 131-124.
توحیدی نژاد فاطمه، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، نجمی نوری عذرا (1393) مقایسه سطوح مختلف بیان ژنRheb  در بافت های مختلف بز کرکی راینی. مجله بیوتکنولوژی کشاورزی 6(4)، 50-35.
جعفری دره در امیر حسین، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، ریاحی مدوار علی (1395) بررسی بیان ژن CIB4  در بافت­های مختلف گوسفند کرمانی با استفاده از Real Time qPCR. مجله پژوهش در نشخوارکنندگان 4(4)، 132-119.
حسنی محمدنبی، اسدی فوزی مسعود، اسمعیلی زاده کشکوئیه علی، محمدآبادی محمدرضا (1389) آنالیز ژنتیکی صفات رشد در بز کرکی راینی با استفاده از مدل حیوانی چند متغیره. مجله علوم دامی ایران 41، 329-323.
عسکری ناهید، باقی زاده امین، محمدآبادی محمدرضا (1389). مطالعه تنوع ژنتیکی در چهار جمعیت بز کرکی راینی با استفاده از نشانگرهای ISSR. مجله ژنتیک نوین 5، 56-49.
عسکری ناهید، محمدآبادی محمدرضا، بیگی نصیری محمدتقی و همکاران (1378). مطالعه تنوع ژنتیکی بز کرکی راینی با استفاده از نشانگرهای ربزماهواره. مجله دانش کشاورزی 18، 161-155.
علینقی­زاده روح­الله، محمدآبادی محمدرضا، زکی­زاده سونیا (1389) چند شکلی اگزون 2 ژن BMP15 در بز قرمز جبال بارز. مجله بیوتکنولوژی کشاورزی 2(1)،80-69.
محمدآبادی محمدرضا (1390) ارتباط چندشکلی ژن IGFBP-3 با صفات کرک در بز کرکی راینی. مجله ژنتیک نوین 7، 120-115.
محمدآبادی محمدرضا (1394) تنوع آللی ژن کالپاستاتین در گوسفند سنجابی. مجله پژوهشهای سلولی و مولکولی (مجله زیست شناسی ایران) 28(3)، 402-395.
محمدآبادی محمدرضا (1398) بیان ژن Dlk1 در بز کرکی راینی با استفاده از Real Time PCR. مجله بیوتکنولوژی کشاورزی 11(3)، 205-191.  
 References
Alinaghizadeh R, Mohammadabadi MR, Zakizadeh S (2010) Exon 2 of BMP15 gene polymorphismin Jabal Barez Red Goat. Agric Biotechnol J 2, 69-80 (In Persian).
Askari N, Baghizadeh A, Mohammadabadi MR (2008) Analysis of the genetic structure of Iranian indigenous Raeni cashmere goat populations using microsatellite markers. Biotechnol 2, 1-4.
Askari N, Baghizadeh A, Mohammadabadi MR (2010) Study of genetic diversity in four populations of Raeini Cashmere goat using ISSR markers. Modern Genet J 5, 49-56 (In Persian).
Askari N, Mohammadabadi MR, Baghizadeh A (2011) ISSR markers for assessing DNA polymorphism and genetic characterization of cattle, goat and sheep populations. Iran J Biotechnol 9, 222-229.
Askari N, Mohammadabadi MR, Beygi Nassiry MT et al. (2009) Study of Genetic Diversity of Raeini Cashmere Goat Based on Microsatellite Markers. Journal of Agricultural Science 18, 155-161 (In Persian).
Baghizadeh A, Bahaaddini M, Mohamadabadi MR, Askari N (2009) Allelic Variations in Exon 2 of Caprine MHC Class II DRB3 Gene in Raeini Cashmere Goat. Am-Eurasian J Agric Environment Sci 6, 454-459.
Bahrampour V, Mohammadabadi MR, Mirzaei HR et al. (2008) Molecular analysis of Calpastatin gene in Kermani sheep herds. Journal of Agricultural Sciences and Natural Resources 1, 124-131 (In Persian).
Barazandeh A, Moghbeli SM, Vatankhah M, Mohammadabadi MR (2012) Estimating non-genetic and genetic parameters of pre-weaning growth traits in Raini Cashmere goat. Trop Anim Health Prod 44, 811-817.
Barnoy S, Glaser T, Kosower NS (1997) Calpain and calpastatin in myoblast differentiation and fusion: ef­fects of inhibitors. Biochimica et Biophysica Acta 1358, 181–188.
Bouhrara M, Clerjon S, Damez J et al. (2011). Dynamic MRI and thermal simulation to interpret deformation and water transfer in meat during heating. J Agric Food Chem 59, 1229-1235.
Bouhrara M, Lehallier B, Clerjon S et al. (2012) Mapping of muscle deformation during heating: in situ dynamic MRI and nonlinear registration. Magn Reson Imaging 30, 422–430.
Chua BT, Guo K, Li P (2000) Direct cleavage by the calcium-activated protease calpain can lead to inactivation of caspases. J Biol Chem 275, 5131-5135.
Farr C (2011) Calpain and Calpastatin in a Mouse Model of Acute Myeloid Leukemia. Dissertation, University of Toronto
Geesink, G. H., Taylor, R. G., and Koohmaraie, M. (2005). Calpain 3/p94 is not involved in postmortem proteolysis. J. Anim. Sci. 83, 1646–1652.
Gholamhoseini Gooki F, Mohammadabadi MR, Asadi Fozi M (2018) Polymorphism of the growth hormone gene and its effect on production and reproduction traits in goat. Iran J Appl Anim Sci 8, 653-659.
Gooki FG, Mohammadabadi M, Fozi MA, soflaei M (2019) Association of Biometric Traits with Growth Hormone Gene Diversity in Raini Cashmere Goats. Walailak J Sci Technol 16, 499-508.
Hassani MN, Asadi Fozi M, Esmailizadeh AK, Mohammadabadi MR (2010). A genetic analysis of growth traits in Raieni Cashmere goat using multivariate animal model. Iran J Anim Sci 41, 323-329 (In Persian).
Jafari Darehdor AH, Mohammadabadi MR, Esmailizadeh AK, Riahi Madvar A (2016) Investigating expression of CIB4 gene in different tissues of Kermani Sheep using Real Time qPCR. J Rumin Res 4, 119-132 (In Persian).
Juszczuk-Kubiak E, Wyszy´nska-Koko J, Wici´nska K, Rosochacki S (2008) A novel polymorphism in intron 12 of the bovine calpastatin gene. Mol Biol Rep 35, 29–35.
Kidd VJ, Lahti JM, Teitz T (2000) Proteolytic regulation of apoptosis. Cell Dev Biol 11, 191-201
Khorshidi M, Mohammadabadi MR, Esmailizadeh AK et al. (2019) Comparison of artificial neural network and regression models for prediction of body weight in Raini Cashmere goat. Iran J Appl Anim Sci 9, 453-461.
Koohmaraie M (1996) Biochemical factors regulating the toughening and tenderization process of meat. Meat Sci 43, S193–S201.
Leal-Gutiérrez JD, Elzo MA, Johnson DD et al. (2018) Association of m-Calpain and Calpastatin Polymorphisms with Meat Tenderness in a Brahman–Angus Population. Front Genet 9, 00056e.
Liang B, Duan BY, Zhou XP et al. (2010) Calpain activation promotes BACE1 expression, amyloid precursor protein processing, and amyloid plaque formation in a transgenic mouse model of Alzheimer disease. J Biol Chem 285, 27737–27744.
McGinnis KM, Gnegy ME, Park YH et al. (1999) Procaspase-3 and poly (ADP) ribose polymerase (PARP) are calpain substrates. Biochem Biophys Res Commun 263, 94-99.
Moghadaszadeh M, Mohammadabadi MR, Esmailizadeh Koshkoieh A (2015) Association of Exon 2 of BMP15 Gene with the Litter Size in the Raini Cashmere Goat. Genet the 3rd Millennium 13, 4062-4067.
Mohammadabadi MR (2012) Relationships of IGFBP-3 gene polymorphism with cashmere traits in Raini Cashmere goat. Modern Genet J 7, 115-120 (In Persian).
Mohammadabadi MR (2015) Allelic Diversity of Calpastatin Gene in Sanjabi Sheep. J Mol Cell Res (Iran J Biol) 28, 395-402 (In Persian).
Mohammadabadi MR (2019) Dlk1 gene expression in Raini Cashmere goat using Real Time PCR. Agricultural Biotechnology Journal 11, 191-205 (In Persian).
Mohammadabadi MR, Jafari AHD, Bordbar F (2017) Molecular analysis of CIB4 gene and protein in Kermani sheep. Brazil J Medic Biol Res 5, e6177.
Mohammadabadi MR, Tohidinejad F (2017) Charachteristics determination of Rheb gene and protein in Raini Cashmere goat. Iran J Appl Anim Sci 7, 289-295.
Molaei Moghbeli S, Barazandeh A, Vatankhah M, Mohammadabadi MR (2013) Genetics and non-genetics Parameters of body Weight for Post WEANING Traits in Raini Cashmere goats. Trop Anim Health Prod 45, 1519-24.
Nakagawa T, Yuan J (2000) Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol 150, 887–894.
Noori AN, Behzadi MRB, Mohammadabadi MR (2017) Expression pattern of Rheb gene in Jabal Barez Red goat. Indian J Anim Sci 87, 1375-1378.
Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30, e36.
Ruiz-vela A, Serrano F, Gonzalez MA et al. (2001) Transplanted long-term cultured pre-B1 cells expressing calpastatin are resistant to B cell receptor-induced apoptosis. J Exp Med 194, 247–254.
Shamsalddini S, Mohammadabadi MR, Esmailizadeh AK (2016) Polymorphism of the prolactin gene and its effect on fiber traits in goat. Russ J Genet 52, 461–465.
Tang R, Yu B, Zhang K et al. (2010) Effects of nutritional level on pork quality and gene expression of micro-calpain and calpastatin in muscle of finishing pigs. Meat Science 85, 768–771.
Tohidi nezhad F, Mohammadabadi MR, Esmailizadeh AK, Najmi Noori A (2015) Comparison of different levels of Rheb gene expression in different tissues of Raini Cashmir goat. Agric Biotechnol J 6, 35-50 (In Persian).
Van Ba H, Reddy BV, Hwang I (2015) Role of calpastatin in the regulation of mRNA expression of calpain, caspase, and heat shock protein systems in bovine muscle satellite cells. In Vitro Cell Dev Biol—Anim 51, 447-454.
Wang D, Zhou L, Zhou H et al. (2015) Effects of Nutritional Level of Concentrate‐Based Diets on Meat Quality and Expression Levels of Genes Related to Meat Quality in Hainan Black Goats. Anim Sci J 86,166-173.
Zheng YC, Lin YQ, Yue Y et al. (2011) Expression profiles of myostatin and calpastatin genes and analysis of shear force and intramuscular fat content of yak longissimus muscle. Czech J Anim Sci 56, 544–550.