Leptin gene expression in subcutaneous adipose tissue of Holstein dairy cattle using Real Time PCR

Document Type : Research Paper

Authors

1 PhD Student, Department of Animal Science, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran

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

3 Associate Professor, Department of Animal Science, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

4 Professor, Department of Animal Science, College of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

5 Faculty of Agriculture, Razi University of Kermanshah, Kermanshah, Iran.

Abstract

Objective
In dairy cattle, the increase in milk yield has been accompanied by a more negative energy balance during early lactation and a decrease in fertility. As the hormone leptin is involved in regulation of nutritional status and reproductive function, this hormone is an interesting protein to investigate during the periparturient period in dairy cattle when many changes take place both in energy metabolism and reproductive physiology. The aim of this research was to study leptin gene expression in adipose tissue of Holstein dairy cattle.
 
Materials and Methods
Tissue sampling from subcutaneous adipose tissue of 20 Holstein cattle on the 20th day of lactation, with the same gestational age (second pregnancy) and mean body weight of 680 ± 80 kg was performed and RNA was extracted. Extracted RNA were immediately stored at -80°C.The Quality and quantity of RNA were evaluated and cDNA was synthesized and Real Time PCR was performed. PCR Products were electrophoresed on 2% agarose gel and were evaluated level of gene expression in the studied tissue.
 
Results
Results showed that the leptin gene was expressed in the subcutaneous adipose tissue. These results may show that leptin plays a particular role in fat metabolism.
 
Conclusions
During the periparturient period the cow mobilizes her fat reserves (i.e. adipose tissue) and it appears that all her energy is going to the production of milk, and that other processes like reproduction and immunity, get a lower priority. Because fertility, just as milk production, is also an economically important trait, fertility should be considered as part of a dairy cattle breeding program. Generally, further studies are needed to clarify role of leptin in the physiology of fat metabolism and other materials. This would help us to better understand the mechanisms for the known effect of nutritional factors and body fatness on various functions.

 
Ahsani MR, Mohammadabadi MR, Asadi Fozi M, Esmailizadeh Koshkooieh A, Khezri A, Esmaeili HR (2019) Leptin gene expression in subcutaneous adipose tissue of Holstein dairy cattle using Real Time PCR. Agricultural Biotechnology Journal 11 (1), 135-150. 
 
Agricultural Biotechnology Journal 11 (1), 135-150.
DOI: 10.22103/jab.2019.13778.1126
Received:  February 11, 2019; Accepted: May 10, 2019
© Faculty of Agriculture, Shahid Bahonar University of Kerman-Iranian Biotechnology Society

Keywords


پسندیده مجید، خراتی کوپایی حامد، محمدآبادی محمدرضا، اسماعیلی زاده کشکوئیه علی (1395) ارتباط آلل‏های ژن‏های OPN و PPARGC1A با تعداد سلول‏های بدنی شیر در گاوهای هلشتاین ایران. مجله ژنتیک نوین 11(3)، 365-357.
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.
خراتی کوپایی حامد، محمدآبادی محمدرضا، انصاری مهیاری سعید، اسماعیلی زاده کشکوئیه علی، ترنگ علیرضا، نیکبختی مهدی (1390) بررسی تنوع ژنتیکی جایگاه ژن DGAT1 و میزان ارتباط آن با تولید شیر در جمعیت گاو های هلشتاین ایران.مجله پژوهش­های علوم دامی ایران 3(2)، 192-185.  
خراتی کوپایی حامد، محمدآبادی محمدرضا، ترنگ علیرضا، خراتی کوپایی محمود، اسماعیلی زاده کشکوئیه علی (1391) بررسی ارتباط چند شکلی آللی ژن DGAT با بیماری ورم پستان در جمعیت گاوهای هلشتاین ایران. مجله ژنتیک نوین 7(1)، 104-101.
محمدآبادی محمدرضا، کرد محبوبه، نظری محمود (1397) مطالعه بیان ژن لپتین در بافت‌های مختلف گوسفند کرمانی با استفاده از Real Time PCR. مجله بیوتکنولوژی کشاورزی 10(3)، 122-111.
محمدآبادی محمدرضا، محمدی اکرم (1389) بررسی ژنوتیپ های بتالاکتوگلوبولین در گاوهای بومی و هلشتاین استان کرمان. مجله تولیدات دامی 12(2)، 67-61.
References
Alinaghizadeh R, Mohammad Abadi MR, Moradnasab Badrabadi S (2007) Kappa-casein gene study in Iranian Sistani cattle breed (Bosindicus) using PCR-RFLP. Pak J Biol Sci, 10, 4291-4294.
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.
Barazandeh A, Mohammadabadi MR, Ghaderi M, Nezamabadipour H (2016) Predicting CpG Islands and Their Relationship with Genomic Feature in Cattle by Hidden Markov Model Algorithm. Iran J App Anim Sci 6, 571-579.
Bartha T, Sayed Ahmed A, Rudas P (2005) Expression of leptin and its receptors in various tissues of ruminants. Domest Anim Endocrinol 29, 193-202.
Batista AM, Silva DMF, Rêgo MJBM et al. (2013) The expression and localization of leptin and its receptor in goat ovarian follicles. Anim Reprod Sci 141, 142-147.
Bocquier F, Bonnet M, Faulconnier Y et al. (1998) Effect of photoperiod and feeling level on perirental adipose tissue metabolic activity and leptin synthesis in the ovariectomized ewe. Reprod Nut Develop 38, 489-498. 
Bonnet M, Gourdou I, Leroux C et al. (2002) Leptin expression in the ovine mammary gland: Putative sequential involvement of adipose, epithelial, and myoepithelial cells during pregnancy and lactation. J Anim Sci 80, 723–728.
Chilliard Y, Bonnet M, Delavaud C et al. (2001) Leptin in ruminants. Gene expression in adipose tissue and mammary gland regulation of plasma concentration. Domest Anim Endocrinol 21, 271-295.
Cioffi JA, Van Blerkom J, Antczak M et al. (1997) The expression of leptin and its receptors in pre-ovulatoryhuman follicles. Mol Human Reprod 3, 467–472.
Dyer CJ, Simmons JM, Matteri RL et al. (1997) cDNA cloning and tissue specific gene expression of ovine leptin, NPY-Y1 receptor and NPY-Y2 receptor. Domest Anim Endocrynol 14, 295-303.
Ebrahimi Z, Mohammadabadi MR, Esmailizadeh AK et al. (2015a) Association of PIT1 gene with milk fat percentage in Holstein cattle. Iran J Appl Anim Sci 5, 575-582.
Ebrahimi Z, Mohammadabadi MR, Esmailizadeh AK (2015b) Association of PIT1 gene and milk protein percentage in Holstein cattle. J Livest Sci Technol 3, 41-49.
Ghasemi M, Baghizadeh A, Abadi MRM (2010) Determination of genetic polymorphism in Kerman Holstein and Jersey cattle population using ISSR markers. Aust J Basic Appl Sci 4, 5758-5760.
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).
Javanmard A, Mohammadabadi MR, Zarrigabayi GE et al. (2008) Polymorphism within the intron region of the bovine leptin gene in Iranian Sarabi cattle (Iranian Bos taurus). Russ J Genet 44, 495-497.
Kharrati Koopaei H, Mohammad Abadi MR, Ansari Mahyari S et al. (2012a) Effect of DGAT1 variants on milk composition traits in Iranian Holstein cattle population. Anim Sci Papers Report 30, 231-240.
Kharrati Koopaei H, Mohammadabadi MR, Ansari Mehyari S et al. (2011) Genetic Variation of DGAT1 Gene and its Association with Milk Production in Iranian Holstein Cattle Breed Population. Iran J Anim Sci Res 3, 185-192 (in Persian).
Kharrati koopaei H, Mohammadabadi MR, Tarang A et al. (2012b) Study of the association between the allelic variations in DGAT1 gene with mastitis in Iranian Holstein cattle. Modern Genet J 7, 101-104 (in Persian).
Liefers SC, Veerkamp RF (2002) Association between leptin gene polymorphism and production, live weight energy balance, feed intake and fertility in Holstein heifers. J Dairy Sci 85, 1633–1638.
Mohammad Abadi MR, Mohammadi A (2010) Study of beta-lactoglobulin genotypes in native and Holstein cattle of Kerman province. J Anim Prod 12, 61-67 (in Persian).
Mohammadabadi M R, Shaikhaev GO, Sulimova GE et al. (2004) Detection of bovine leukemia virus proviral DNA in Yaroslavsl, Mongolian and black pied cattle by PCR. Cell Mol Biol Letter 9, 766-768.
Mohammadabadi MR, Soflaei M, Mostafavi H, Honarmand M (2011) Using PCR for early diagnosis of bovine leukemia virus infection in some native cattle. Genet Mol Res 10, 2658-2663.
Mohammadabadi MR, Kovalenko TA, Nasiri MR, Sulimova GE (2005) inter-Simple-Sequence Repeat (ISSR)-PCR for the identification of polymorphism in some native cattle breeds. Proceeding of the 3rd Moscow international congress: Biotechnology: state of the art and prospects of development, Moscow, Russia.
Mohammadabadi MR, Tohidinejad F (2017) Charachteristics determination of Rheb gene and protein in Raini Cashmere goat. Iran J Appl Anim Sci 7, 289-295.
Mohammadabadi MR, Jafari AHD, Bordbar F (2017) Molecular analysis of CIB4 gene and protein in Kermani sheep. Brazil J Med Biol Res 50, e6177.
Mohammadabadi MR, Kord M, Nazari M (2018) Studying expression of leptin gene in different tissues of Kermani Sheep using Real Time PCR. Agric Biotechnol J 10, 111-122 (in Persian).
Munoz-Gutiérrez M, Findlay PA, Adam CL et al. (2005) The ovar-ian expression of mRNAs for aromatase, IGF-I receptor, IGF-bindingprotein-2, -4 and -5, leptin and leptin receptor in cycling ewes afterthree days of leptin infusion. Reprod 130, 869–881.
Ostlund REJr, Yang JW, Klein S, Gingerich R (1996) Relation between plasma leptin concentration and body fat, gender, diet, age, and metabolic covariates. J Clinic Endocrinol Metabol 81, 3909–3913.
Pasandideh M, Kharrati Koopaee H, Mohammad Abadi MR, Esmailizadeh Koshkoiyeh A (2016) Association of the OPN and PPARGC1A Alleles with Milk Somatic Cell Count in Iranian Holstein cattle. Modern Genet J 11, 357-365 (in Persian).
Pasandideh M, Mohammadabadi MR, Esmailizadeh AK, Tarang A (2015) Association of bovine PPARGC1A and OPN genes with milk production and composition in Holstein cattle. Czech J Anim Sci 60, 97-104.
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.
Pisani LF, Antonini S, Pocar P et al. (2008) Effects of pre-mating nutrition on mRNA levels ofdevelopmentally relevant genes in sheep oocytes and granulosa cells. Reprod 136, 303–312.
Ruzina MN, Shtyfurko TA, Mohammadabadi MR et al. (2010) Polymorphism of the BoLA-DRB3 gene in the Mongolian, Kalmyk, and Yakut cattle breeds. Russ J genet 46, 456-463.
Sarkar M, Schilffarth S, Schams D et al. (2010) Theexpression of leptin and its receptor during different physiologicalstages in the bovine ovary. Mol Reprod Devlop 77, 174–181.
Schoof E, Stuppy A, Harig F et al. (2004) Comparison of leptin gene expression in different adipose tissues in children and adults. Europ J Endocrinol 150, 579–584.
Shojaei M, Mohammadabadi MR, Asadi Fozi M et al. (2010) Association of growth trait and Leptin gene polymorphism in Kermani sheep. J Cell Mol Res 2, 67-73.
Sulimova GE, Abani MA, Rostamzadeh J et al. (2007) Allelic polymorphism of kappa-casein gene (CSN3) in Russian cattle breeds and its informative value as a genetic marker. Russ J Genet 43, 88-95.
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.
Yonekura S, Kitade K, Furukawa G et al. (2002) Effects of aging and weaning on mRNA expression of leptin and CCK receptors in the calf rumen and abomasum. Domest Anim Endocrinol 22, 25-35. 
Yonekura S, Senoo T, Kobayashi Y et al. (2003) Effect of acetate and butyrate on the expression of leptin and short form leptin receptor in bovine and rat anterior pituitary cells. General Comparative Endocrinol 133, 165-172.
Yuen BS, Owens PC, McFarlane JR et al. (2002) Circulating leptin concentration are positively related to leptin messenger RNA expression in adipose tissue of fetal sheep in pregnant ewe fed at or below maintenance requirements during late gestation. Biol Reprod 67, 911-916.
Zieba AD, Amstalden M, Morton S et al. (2003) Effects of leptin on basal and GHRH-stimulated GH secretion from the bovine adenohypophysis are dependent upon nutritional status. J Endocrinol 178, 83-89.