The effect of Cannabis seed on DLK1 gene expression in heart tissue of Kermani lambs

Document Type : Research Paper

Authors

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

2 Assistant professor, Animal Science Department, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

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

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

5 Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran

Abstract

Objective
Cannabis seeds have high amounts of crude protein and energy, so they can better resist the microbial digestion of the rumen and pass through the intact rumen to a greater extent and be digested and absorbed in the intestine. This means they can improve livestock performance. Therefore, they can be used for this purpose. On the other hand, DLK1 gene plays an important role in controlling cell differentiation processes, including muscle and nerve differentiation, adipogenesis, hematopoiesis, and bone differentiation throughout fetal and adult life. Therefore, the aim of this study was to investigate the effect of cannabis seeds on DLK1 gene expression in the heart tissue of Kermani lambs.
Materials and Methods
To carry out this research, 12 six-month-old Kermani lambs of equal weight were used in a completely randomized design in three experimental groups.After slaughter, sampling of heart tissue (with 3 repeats) was done from both groups.The samples were quickly placed in liquid nitrogen and then stored in a -80 freezer.Total RNA was extracted using a standard kit.The quality and quantity of extracted RNA was evaluated.cDNA synthesis kit of Fermentase Company was used for cDNA synthesis.Real time PCR reaction using Syber Green method was used to check the relative level of gene expression.To analyze the data obtained from real time PCR, the method of Pfaffl et al. was used.
Results
The ratio of A260/A280 for the extracted RNAs was equal to 1.9-1.8, which indicates their appropriate and desirable quality. The results of real time PCR curves and electrophoresis of PCR products on agarose gel showed that DLK1 gene is expressed in heart tissue. The results of the present study also showed that feeding cannabis at the level of 1% significantly (P<0.05) increases the expression of DLK1 gene in the heart tissue by 4.74 times compared to the control group.
Conclusions
Based on the results of the present study, it can be concluded that cannabis can be used in the diet of sheep to improve the structure of the heart through positive effects on the expression of the DLK1 gene. In addition, the interesting results of the present study in response to the use of Cannabis seeds in the diet of sheep open a new path for wider research in this field.

Keywords

References

بارانی  مهدی،  افضلی نظر، حسینی واشان سید جواد (2017) اثر تغذیه دانه شاهدانه (cannabis sativa L) اکسترود شده بر عملکرد، اجزای لاشه، پاسخ ایمنی همورال و الگوی لیپیدی پلاسما در جوجه­های گوشتی. تحقیقات تولیدات دامی، 6(2)، 39-49. 
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.
جمال پور مهرنسا، دادپسند محمد، آتشی هادی و همکاران (1397) واکاوی بیوانفورماتیکی و فیلوژنی ناحیه 5`UTR ژن نوروپپتید وای (Neuropeptide Y) و ارتباط آن با صفات تولیدی در مرغ‌ بومی فارس. نشریه علوم دامی ایران، 49(3)، 458-453.
کرمشاهی امجزی خدیجه، جلیلوند قاسم، دیانی امید، دهقان بنادکی مهدی (1398) اثرات تغذیه شاهدانه در جیره‌های با پروتئین خام متفاوت بر عملکرد، قابلیت هضم ، فراسنجه‌های شکمبه‌ای و سنتز پروتئین میکروبی در بره‌های پرواری بلوچی. پژوهش در نشخوارکنندگان، 7(2)، 33-58.
محمدآبادی محمدرضا (1398) بیان ژن DLK1 در بز کرکی راینی با استفاده از Real Time PCR. مجله بیوتکنولوژی کشاورزی 11(3)،205-191.
محمدآبادی محمدرضا (1399) بیان ژن ESR1 در بز کرکی راینی با استفاده از real time PCR‎. مجله بیوتکنولوژی کشاورزی 12(1)، 192-177.
محمدآبادی محمدرضا، خیرالدین حمید، لطیفی آیگین، بابنکو اولنا ایوانیونا (1401) پروفایل بیانی ژن  DNAH1در بافت بیضه بز کرکی راینی. مجله بیوتکنولوژی کشاورزی، 14(3)، 243-256.
محمدآبادی محمدرضا، سفلایی محمد (1399). پروفایل بیانی mRNA مختص بافت ژن BMP15 در بز. مجله بیوتکنولوژی کشاورزی 12(3)، 208-191.
محمدآبادی محمدرضا، شبان جرجندی دیانا، عرب پور رق آبادی زهرا، ابارقی فاطمه، ساسان حسینعلی، بردبار فرهاد (1401) نقش رازیانه بر بیان ژن DLK1 در بافت قلب گوسفند. مجله بیوتکنولوژی کشاورزی، 14(2)، 155-133.
محمدی فر آمنه، محمدآبادی محمدرضا (1390) کاربرد نشانگرهای ریزماهواره برای مطالعه ژنوم گوسفند کرمانی. مجله علوم دامی ایران 42(4)، 344-337.
محمودی محمد،  فرهومند پرویز، آذرفر آرش (2012) اثر سطوح مختلف جیره­ای شاهدانه (Cannabis sativa L) بر عملکرد، وزن اندام-های داخلی و میزان کلسترول سرم جوجه­های گوشتی. فصلنامه علمی پژوهشی گیاهان دارویی، 2(42)، 121-129.
محمودی مریم ، آیت اللهی مهرجردی احمد ، محمدآبادی محمدرضا (1396) بررسی اگزون چهارم ژن کاپاکازئین گوسفند کرمانی با تکنیک PCR-RFLP. مجله بیوتکنولوژی کشاورزی، 9(3)، 128-119.
References
Abdallah BM, Jensen CH, Gutierrez G, et al. (2004) Regulation of human skeletal stem cells differentiation by DLK1/Pref-1. J Bone Miner Res 19, 841–852.
Aminafshar M, Bahrampour V, Baghizadeh A, et al. (2014) CD44 gene expression in mature, immature oocytes and fetal kermani, baluchi sheep and rayeni, tali goats. J cell Anim Biol 8 (8), 156-160.
Amiri Roudbar M, Mohammadabadi MR, Mehrgardi AA, Abdollahi-Arpanahi A (2017) Estimates of variance components due to parent-of-origin effects for body weight in Iran-Black sheep. Small Rum Res 149, 1-5.
Amiri Roudbar M., Abdollahi-Arpanahi R., Ayatollahi Mehrgardi A., et al. (2018) Estimation of the variance due to parent-of-origin effects for productive and reproductive traits in Lori-Bakhtiari sheep. Small Rumin Res 160, 95-102.
Bahrampour V, Mohammadabadi MR, Mirzaei HR, et al. (2008) Molecular analysis of Calpastatin gene in Kermani sheep herds. J Agric Sci Nat Resource 15 (4), 124-131.
Barani M, Afzali N, Hosseini-Vashan SJ (2017) Effects of dietary inclusion of extruded hempseed (Cannabis sativa L.) on performance, carcass components, humoral immune response and plasma lipid profile of broiler chickens. Anim Produc Res 6(2) 39-49 (In Persian).
Bujak M, Frangogiannis NG (2007) The role of TGF-beta signaling in myocardial infarction and cardiac remodeling. Cardiovasc Res 74, 184–195.
Callaway JC (2004) Hempseed as a nutritional resource: An overview. Euphytica 140(1-2), 65-72.
Chen L, Qanie D, Jafari A, et al. (2011) Delta-like 1/fetal antigen-1 (DLK1/FA1) is a novel regulator of chondrogenic cell differentiation via inhibition of the Akt kinase-dependent pathway. J Biol Chem 286, 32140–32149.
Driesen RB, Nagaraju CK, Abi-Char J, et al. (2014) Reversible and irreversible differentiation of cardiac fibroblasts. Cardiovasc Res 101, 411–422.
Ferron SR, Charalambous M, Radford E, et al. (2011) Postnatal loss of DLK1 imprinting in stem cells and niche astrocytes regulates neurogenesis. Nature 475, 381–385.
Hendriks H, Malingré TM, Batterman S, Bos R (1975) Mono-and sesqui-terpene hydrocarbons of the essential oil of Cannabis sativa. Phytochem 14, 814-815.
Jamalpour M, Dadpasand M, Atashi H, et al. (2018) Bioinformatics and phylogenetic analysis for 5′ UTR region of neuropeptide Y gene and its association with body weight and egg production traits in Fars native chickens. Iran J Anim Sci 49(3), 453-458 (In Persian).
Karamshahi Amjazi Kh, Jalilvand Gh, Dayani O, Dehghan Banadak M (2019) Effects of feeding hemp seeds in diets with different levels of crude protein on Performance, Digestibility, Ruminal metabolites and Synthesis of Microbial Protein in Baluchi fattening lambs. J Rumin Res 7(2), 33-58 (In Persian).
Kocher O, Kennedy SP, Madri JA (1990) Alternative splicing of endothelial cell fibronectin mRNA in the IIICS region. Functional significance. Am J Pathol 137, 1509–1524.
Laborda J (2000) The role of the epidermal growth factor-like protein dlk in cell differentiation. Histol Histopathol 15, 119–129.
Leeson S, Summers JD, Caston LJ (2001) Response of layers to low nutrient density diets. J Appl Poult Res 10(1), 46-52.
Li RK, Li G, Mickle DA, et al. (1997) Overexpression of transforming growth factor-beta1 and insulinlike growth factor-I in patients with idiopathic hypertrophic cardiomyopathy. Circulation 96, 874–881.
Lottrup G, Nielsen JE, Skakkebæk NE, et al. (2015) Abundance of DLK1, differential expression of CYP11B1, CYP21A2 and MC2R, and lack of INSL3 distinguish testicular adrenal rest tumours from leydig cell tumours. Eur J Endocrinol 172, 491–499.
Lui JC, Finkielstain GP, Barnes KM, Baron J (2008) An imprinted gene network that controls mammalian somatic growth is down-regulated during postnatal growth deceleration in multiple organs. Am J Physiol – Regul Integr Comp Physiol 295, R189-196.
Mahmoodi M, Ayatollahi A, Mohammadabadi MR (2018) Studying exon 4 of kappa-casein gene in Kermani sheep using PCR-RFLP. Agric Biotechol J 9 (3), 119-128 (In Persian).
Mahmoudi M, Farhoomand P, Azarfar A (2012). Eeffect of graded levels of hemp seed (Cannabis Sativa L.) on performance, organ, weight and serum cholesterol levels on broilers. J Med Plants 11(9) 121-129 (In Persian).
Masoudzadeh SH, Mohammadabadi M, Khezri A, et al. (2020a) Effects of diets with different levels of fennel (Foeniculum vulgare) seed powder on DLK1 gene expression in brain, adipose tissue, femur muscle and rumen of Kermani lambs. Small Rumin Res 193, e106276.
Masoudzadeh SH, Mohammadabadi MR, Khezri A, et al. (2020b) DLK1 Gene Expression in Different Tissues of Lamb. Iran J Appl Anim Sci 10 (4), 669-677.
Mohamadipoor Saadatabadi L, Mohammadabadi M, Amiri Z, et al. (2021) Signature selection analysis reveals candidate genes associated with production traits in Iranian sheep breeds. BMC Vet Res 17 (1), 1-9
Mohammadabadi M, Masoudzadeh SH, Khezri A, et al. (2021) Fennel (Foeniculum vulgare) seed powder increases Delta-Like Non-Canonical Notch Ligand 1 gene expression in testis, liver, and humeral muscle tissues of growing lambs. Heliyon 7 (12), e08542.
Mohammadabadi M, Shaban Jorjandy D, Arabpoor Raghabadi Z, et al. (2022) The role of fennel on DLK1 gene expression in sheep heart tissue. Agric Biotechnol J 14 (2), 155-170 (In Persian)
Mohammadabadi M, Soflaei M (2020) Tissue-specific mRNA expression profile of BMP15 gene in goat. Agric Biotechnol J 12, 191-208 (In Persian).
Mohammadabadi MR (2016) Role of clostridium perfringens in pathogenicity of some domestic animals. J Adv Agric 7 (3), 1117-1121.
Mohammadabadi MR (2017) Inter-Simple Sequence Repeat loci Associations with Predicted Breeding Values of Body Weight in Kermani Sheep. Genet 3rd Millen 14 (4), 4383-4390.
Mohammadabadi MR (2019) Dlk1 gene expression in Raini Cashmere goat using Real Time PCR. Agric Biotechnol J 11 (3), 191-205 (In Persian).
Mohammadabadi MR (2020) Expression of ESR1 gene in Raini Cashmere goat using real time PCR. Agric Biotechnol J 12 (1), 177-192 (In Persian).
Mohammadabadi MR, Esfandyarpoor E, Mousapour A (2017) Using Inter Simple Sequence Repeat Multi-Loci Markers for Studying Genetic Diversity in Kermani Sheep. J Res Dev 5 (2), e154.
Mohammadabadi MR, Kheyrodin H, Latifi A, Babenko OI (2022) mRNA expression profile of DNAH1 gene in testis tissue of Raini Cashmere goat. Agric Biotechnol J 14 (3), 243-256.
Mohammadifar A, Mohammadabadi MR (2011) Application of Microsatellite Markers for a Study of Kermani Sheep Genome. Iran J Anim Sci 42 (4), 337-344 (In Persian).
Oczkowicz M, Piestrzyska-Kajtoch A, Piórkowska K, et al. (2010) Expression of DLK1 and MEG3 genes in porcine tissues during postnatal development. Genet Mol Biol 33, 790-794.
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.
Rocha ST, Tevendale M, Knowles E, et al. (2007) Restricted co-expression of DLK1 and the reciprocally imprinted non-coding RNA, Gtl2: Implications for cis-acting control. Dev Biol, 306, 810–823.
Rodriguez P, Sassi Y, Troncone L, et al. (2019) Deletion of delta-like 1 homologue accelerates fibroblast–myofibroblast differentiation and inducesmyocardial fibrosis. Eur Heart J 40, 967–978.
Safaei SMH, Dadpasand M, Mohammadabadi M, et al. (2022) An Origanum majorana Leaf Diet Influences Myogenin Gene Expression, Performance, and Carcass Characteristics in Lambs. Animals 13 (1), e14.
Sakajiri S, O’Kelly J, Yin D, et al. (2005) DLK1 in normal and abnormal hematopoiesis. Leukemia 19, 1404–1410.
Serini G, Bochaton-Piallat ML, Ropraz P, et al. (1998) The fibronectin domain ED-A is crucial for myofibroblastic phenotype induction by transforming growth factor-beta1. J Cell Biol 142, 873–881.
Shahsavari M, Mohammadabadi M, Khezri A, et al. (2021) Correlation between insulin-like growth factor 1 gene expression and fennel (Foeniculum vulgare) seed powder consumption in muscle of sheep. Animal Biotechnology 33, 1-11.
Small E, Cronquist A (1976) A practical and natural taxonomy for Cannabis. Taxon, 405-435.
Tanimizu N, Nishikawa M, Saito H, et al. (2003) Isolation of hepatoblasts based on the expression of dlk/pref-1. J Cell Sci 116, 1775-1786.
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).
Travers JG, Kamal FA, Robbins J, et al. (2016) Cardiac fibrosis: the fibroblast awakens. Circ Res 118, 1021–1040.
Wang XS, Tang CH, Yang XQ, Gao WR (2008) Characterization, amino acid composition and in vitro digestibility of hemp (Cannabis sativa L.) proteins. Food Chem 107(1), 11-18.
Yevtodiyenko A, Schmidt JV (2006) DLK1 expression marks developing endothelium and sites of branching morphogenesis in the mouse embryo and placenta. Dev Dyn 235, 1115–1123.
Agricultural Biotechnology Journal
Volume 15, Issue 1
April 2023
Pages 217-234

Files

Share

How to cite

Statistics