INVESTIGATION OF G+265C AND G-1539A SINGLE NUCLEOTIDE POLYMORPHISMS OF TOLL-LIKE RECEPTOR 4 GENE (TLR4) IN SOME CATTLE BREEDS RAISED IN TURKEY

Authors

  • Özgecan Korkmaz Ağaoğlu Burdur Mehmet Akif Ersoy University Faculty of Veterinary Medicine, Department of Animal Science, Burdur, Turkey http://orcid.org/0000-0002-7414-1725
  • Bilal Akyüz Erciyes University, Faculty of Veterinary Medicine, Department of Genetics, Kayseri, 38039, Turkey http://orcid.org/0000-0001-7548-9830
  • Emel Zeytünlü Mehmet Akif Ersoy University, Institute of Health Sciences, Department of Animal Science, Burdur, 15030, Turkey
  • Ali Reha Ağaoğlu Burdur Mehmet Akif Ersoy University Faculty of Veterinary Medicine, Department of Obstetrics and Gynecology, Burdur, 15030, Turkey http://orcid.org/0000-0002-6545-8800

DOI:

https://doi.org/10.26873/SVR-914-2020

Abstract

Toll-like receptors play an essential role in how the innate immune system reacts to pathogens. These receptors help the innate immune system recognise the antigenic structure of pathogens and initiate the inflammatory response. TLR4 is one of these receptors, and it has been identified as a candidate molecular marker for resistance to mastitis in cattle. This study aimed to describe G-1539A (BgII) in promoter and G+265C (MspI) in 5'-UTR polymorphisms in the TLR4 gene using Zavot (n=60), East Anatolian Red (EAR, n=49), Anatolian Black (AB, n=59), South Anatolian Red (SAR, n=42), Turkish Gray (TG, n=60) and Holstein (n=218) cattle breeds. For this purpose, we used polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) to genotype a total of 488 cattle. Two alleles (A and G) and three genotypes (GG, GA and AA) were examined by digestion of the PCR product with BgII restriction enzyme. The frequency of the G allele was higher in all investigated breeds except Zavot. The GA genotype was found to be the most common genotype in the EAR, SAR, TG, AB, Holstein, and Zavot breeds. Two alleles (C and G) and three genotypes (GG, GC and CC) were examined by digestion of the PCR product for the G+265C SNP with the MspI restriction enzyme. The most prevalent in terms of frequency was the C allele in all breeds with regard to G+265C SNP. All examined breeds were within the Hardy-Weinberg equilibrium (p>0.05). Consequently, the existence of two SNPs of the TLR4 gene has been reported for the first time in six cattle breeds raised in Turkey. The study demonstrated that the investigated breeds continue to exhibit variation in terms of these two SNPs. It might be possible to utilise these polymorphisms in efforts to breed herds resistant to significant breeding diseases, such as mastitis.

Key words: cattle; PCR-RFLP; promoter; SNP; TLR4

RAZISKOVANJE ENOJNIH NUKLEOTIDNIH POLIMORFIZMOV G+265C IN G-1539A GENA RECEPTORJA TLR4 PRI NEKATERIH PASMAH KRAV, GOJENIH V TURČIJI

 

Povzetek: Tolični receptorji oz. receptorji TLR (iz angl. Toll-like receptors) igrajo bistveno vlogo pri reakciji prirojenega imunskega sistema na patogene. Ti receptorji pomagajo prirojenemu imunskemu sistemu prepoznati antigensko strukturo patogenov in sprožiti vnetni odziv. TLR4 je bil izmed skupine toličnih receptorjev prepoznan kot kandidatni molekularni označevalec za odpornost na mastitis pri govedu. V študiji smo želeli opisati G-1539A (BgII) v promotorju in G+265C (MspI) v 5'-UTR polimorfizmih gena TLR4 pri različnih pasmah goveda: pasmi Zavot (n = 60), vzhodno anatolskem rdečem govedu (EAR, n = 49), anatolijskemu črnemu govedu (AB, n = 59), južno anatolskemu rdečemua govedu (SAR, n = 42), turškem sivem govedu (TG, n = 60) in govedu pasme Holstein (n = 218). V ta namen smo uporabili verižno reakcijo s polimerazo in polimorfizem dolžin restrikcijskih odsekov (PCR-RFLP) za genotipizacijo 488 govedi. Z restrikcijsko cepitvijo produktov PCR z restrikcijskim encimom BgII smo pregledali dva alela (A in G) in tri genotipe (GG, GA in AA). Pogostost alela G je bila večja pri vseh raziskanih pasmah, razen pasmi Zavot. Ugotovljeno je bilo, da je genotip GA najpogostejši genotip pri pasmah goveda EAR, SAR, TG, AB, Holstein in Zavot. Dva alela (C in G) in trije genotipi (GG, GC in CC) so bili pregledani z restrikcijsko cepitvijo produkta PCR za SNP G+65C z restrikcijskim encimom MspI. Alel C je bil najbolj razširjen pri vseh pasmah. Vse pregledane pasme so bile znotraj Hardy-Weinbergovega ravnotežja (p > 0,05). V raziskavo torej poročamo prvič o obstoju dveh SNP gena TLR4 pri šestih pasmah govedi, vzrejenih v Turčiji. Študija je pokazala, da so pri preiskanih pasmah vidne razlike v teh dveh SNP. Te polimorfizme bi bilo mogoče uporabiti pri prizadevanju za vzrejo čred, odpornih na pomembne bolezni pri govedu, kot je mastitis.

Ključne besede: govedo; PCR-RFLP; promotor; SNP; TLR4

Author Biography

Özgecan Korkmaz Ağaoğlu, Burdur Mehmet Akif Ersoy University Faculty of Veterinary Medicine, Department of Animal Science, Burdur, Turkey

Department of Animal Science,

Assoc. Prof. Dr.

References

(1.) Jones WP, Hansen LB, Chester-Jones H. Response of health care to selection for milk yield of dairy cattle. J Dairy Sci 1994; 77: 3137–52.

(2.) Lucy MC. Reproductive loss in high-producing dairy cattle: where will it end? J Dairy Sci 2001; 84: 277–93.

(3.) Egger-Danner C, Willam A, Fuerst C, Schwarzenbacher H, Fuerst-Waltl B. Hot topic: effect of breeding strategies using genomic information on fitness and health. J Dairy Sci 2012; 95: 4600–9.

(4.) Parker Gaddis KL, Cole JB, Clay JS, Maltecca C. Genomic selection for producer-recorded health event data in US dairy cattle. J Dairy Sci 2014; 97: 3190–9.

(5.) Takeda K, Akira S. Roles of toll-like receptors in innate immune responses. Genes Cells 2001; 6: 733–42.

(6.) Takeda K, Kaisho T, Akira S. Toll-like receptors. Annu Rev. Immunol 2003; 21: 335–376.

(7.) Kaisho T, Akira S. Toll-like receptor function and signaling. J Allergy Clin Immunol 2006; 117: 979–87.

(8.) Medzhitov R, Janeway C. Innate immunity. N Engl J Med 2000; 343: 338–44.

(9.) Akira S, Hemmi H. Recognition of pathogen-associated molecular patterns by TLR family. Immunol Lett 2003; 85: 85–95.

(10.) Mucha R, Bhide MR, Chakurkar EB, Novak M, Mikula IS. Toll-like receptors TLR1, TLR2 and TLR4 gene mutations and natural resistance to Mycobacterium avium subsp. paratuberculosis infection in cattle. Vet Immunol Immunopathol 2009; 128: 381–8.

(11.) Coll RC, O'Neill LA. New insights into the regulation of signaling by Toll-like receptors and Nod-like receptors. J Innate Immun 2010; 2: 406–21.

(12.) Wang X, Xu S, Gao X, Ren H, Chen J. Genetic polymorphism of TLR4 gene and correlation with mastitis in cattle. J Genet Genomics 2007; 34(5): 406–12.

(13.) Akira S, Takeda K, Kaisho T. Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immun 2001; 2(8): 675–80.

(14.) Akira S, Takeda KT. Toll-like receptor signaling. Nat Rev Immunol 2004; 4(7): 499–511.

(15.) Hawn TR, Misch EA, Dunstan, SJ, et al. A common human TLR1 polymorphism regulates the innate immune response to lipopeptides. Eur J Immunol 2007; 37(8): 2280–9.

(16.) Omueti KO, Mazur DJ, Thompson KS, Lyle EA, Tapping RI. The polymorphism P315L of human Toll-like receptor 1 impairs innate immune sensing of microbial cell wall components. J Immunol 2007; 178: 6387–94.

(17.) White SN, Kata SR, Womack JE. Comparative fine maps of bovine toll-like receptor 4 and toll-like receptor 2 regions. Mamm Genome 2003; 14(2): 149–55.

(18.) Schwartz DA. The genetics of innate immunity. Chest 2002; 121: 62S–8S.

(19.) Sharma BS, Leyva I, Schenkel F, Karrow NA. Association of toll-like receptor 4 polymorphisms with somatic cell score and lactation persistency in Holstein bulls. J Dairy Sci 2006; 89(9): 3626–35.

(20.) Pinedo PJ, Buergelt CD, Donovan GA, et al. Candidate gene polymorphisms (BoIFNG, TLR4, SLC11A1) as risk factors for paratuberculosis infection in cattle. Prev Vet Med 2009; 91(2/4): 189–96.

(21.) Pulido I, Leal M, Genebat M, Pacheco YM, Sáez ME, Soriano-Sarabia N. The TLR4 ASP299GLY polymorphism is a risk factor for active tuberculosis in Caucasian HIV-infected patients. Curr HIV Res 2010; 8(3): 253–8.

(22.) Prakash O, Kumar A, Sonwane A, et al. Polymorphism of cytokine and innate immunity genes associated with bovine brucellosis in cattle. Mol Biol Rep 2014; 41(5): 2815–25.

(23.) Goldammer T, Zerbe H, Molenaar A, et al. Mastitis increases mammary mRNA abundance of a-defensin 5, toll-like receptor 2 (TLR2), and TLR4 but not TLR9 in cattle. Clin Vaccine Immunol 2004; 11: 174–85.

(24.) Zhang LP, Gan QF, Ma TH, et al. Toll-like receptor 2 gene polymorphism and its relationship with SCS in dairy cattle. Anim Biotechnol 2009; 20(3): 87–95.

(25.) Yuan Z R, Li J, Zhang, LP, et al. Novel SNPs polymorphism of bovine CACNA2D1 gene and their association with somatic cell score. Afr J Biotechnol 2011; 10(10): 1789–93.

(26.) Li Z, Zhang H, Wang H, et al. Polymorphism in the promoter of TLR4 gene by PCR-RFLP and its association with somatic cell score in Chinese Holstein. Arch Tierzucht 2014; 57(6):1–6.

(27.) Yeh FC, Yang RC, Boyle T. POPGENE version 1.32: Microsoft Windows–based freeware for population genetic analysis. Edmonton, Alberta, Canada : Center for International Forestry Research, University of Alberta, 2000.

(28.) Rezazadeh M, Hajilooi M, Rafiei A, et al. TLR4 polymorphism in Iranian patients with brucellosis. J Infect 2006; 53(3): 206–10.

(29.) Wang XP, Xu SZ, Gao X, Li JY. Cloning and SNP screening of the TLR4 gene and the association between its polymorphism and somatic cell score in dairy cattle. S Afr J Anim Sci 2008; 38(2): 101–9.

(30.) Mariotti M, Williams JL, Dunner S, et al. Polymorphisms within the toll-like receptor (TLR)-2, -4, and -6 genes in cattle. Diversity 2009; 1: 7–18.

Downloads

Additional Files

Published

2020-03-31

How to Cite

Korkmaz Ağaoğlu, Özgecan, Akyüz, B., Zeytünlü, E., & Ağaoğlu, A. R. (2020). INVESTIGATION OF G+265C AND G-1539A SINGLE NUCLEOTIDE POLYMORPHISMS OF TOLL-LIKE RECEPTOR 4 GENE (TLR4) IN SOME CATTLE BREEDS RAISED IN TURKEY. Slovenian Veterinary Research, 57(1). https://doi.org/10.26873/SVR-914-2020

Issue

Section

Original Research Article