Identification of Genetic Variation of Melatonin Receptor 1A (MTNR1A) Gene in Kıvırcık Breed Ewes by MnlI and RsaI Restriction Enzymes

Melatonin reseptör 1A (MTNR1A) geni koyunlarda mevsime bağlı üreme fonksiyonlarını düzenleyen bir hormon olan melatonini kodlamaktadır. Bu çalışmanın amacı Kıvırcık ırkı koyunda MTNR1A geninin genetik varyasyonunun ve allel çeşitliliğini belirlemektir. Kırklareli ve İstanbul illerindeki 5 farklı çiftlikten olmak üzere toplam 110 adet Kıvırcık ırkı koyununa ait kan örnekleri toplanarak DNA izolasyonu yapılmıştır. MTNR1A geninde polimorfik olan ekzon 2 bölgesi PZR ile yükseltgenmiş olup MnlI ve RsaI enzimleri kullanılarak allel ve genotip tespitleri yapılmıştır. Gözlenen alleller ve genotipler MnlI için M (0.891) ve m (0.109) alleleri ile MM (0.782) ve Mm (0.218) genotipleri, RsaI için C (0.682) ve T (0.318) alleleri ile CC (0.582), CT (0.200) ve TT (0.218) genotipleri olmuştur. Kıvırcık koyunlarında mm genotipi gözlenmemiş olup, en yüksek oranda gözlenen genotipler, koyunlarda mevsim dışı üreme faaliyetlerini pozitif olarak etkilediği bilinen MM (%78) ve CC (%58) olarak tespit edilmiştir. Büyük bir çoğunluğu MM ve CC genotiplerine sahip olan Kıvırcık ırkı koyunların mevsim dışı kuzulatmada yaygın olarak kullanılması yetiştiricilere önerilebilir.

Kıvırcık Irkı Koyunlarda Melatonin Reseptör 1A (MTNR1A) Geninin MnlI ve RsaI Restriksiyon Enzimleri ile Genetik Varyasyonunun Belirlenmesi

Melatonin receptor 1A (MTNR1A) gene encodes melatonin hormone which regulates the function of seasonal reproductive activity in sheep. The aim of this study was to make the genetic characterization and identify the variant alleles of MTNR1A gene in Kıvırcık breed. Blood samples of 110 Kıvırcık sheep were collected from five different farms located in Kırklareli and Istanbul. DNA extraction was performed from blood samples. Exon 2, the polymorphic region of Melatonin receptor 1A gene, was amplified and PCR products were genotyped by using MnlI and RsaI enzymes. The observed alleles and genotypes for MnlI enzyme were; M (0.891), m (0.109) and MM (0.782), Mm (0.218) respectively. Kıvırcık sheep was null from mm genotype. Also identified alleles were C (0.682), T (0.318) and genotypes were CC (0.582), CT (0.200), TT (0.218) for RsaI enzyme. The most frequent genotypes were MM (78%) and CC (58%) in Kıvırcık ewes. Since MM and CC genotypes were known with their positive effect on out of season reproductive activities, Kıvırcık ewes with these genotypes might suggested to be used in out of season lambing when demanded.

PDF

___

Yılmaz A, Ekiz B, Özcan M, Kaptan C, Hanoglu H, Erdogan I, Kocak O: Carcass traits of improved and indigenous lamb breeds of north- western Turkey under an intensive production system. Ital J Anim Sci, 8, 667, 2009. DOI: 10.4081/ijas.2009.663
Yalçın BC: Sheep and goats in Turkey. FAO Animal Production and Health Paper, 60, Rome, Italy, 1986.
Mura MC, Luridiana S, Daga C, Bini PP, Carcangiu V: Genotype at the MTNR1A locus and response to melatonin treatment in Sarda lambs. Ital J Anim Sci, 8 (Suppl. 2): 114-116, 2009. DOI: 10.4081/ijas.2009.s2.114
Malpaux B, Migaud M, Tricoire H, Chemineau P: Biology of mammalian photoperiodism and the critical role of the pineal gland and melatonin. J Biol Rhythms, 16, 336-347, 2001. DOI: 10.1177/074873001129002051
Bittman EL, Dempsey RJ, Karsch FJ: Pineal melatonin secretion drives the reproductive response to daylength in the ewe. Endocrinology, 113, 2283, 1983. DOI: 10.1210/endo-113-6-2276
Malpaux B, Daveau A, Maurice-Mandon F, Duarte G, Chemineau P: Evidence that melatonin acts in the premammillary hypothalamic area to control reproduction in the ewe: Presence of binding sites and stimulation of luteinizing hormone secretion by in situ microimplant delivery. Endocrinology, 139, 1508-1516, 1998. DOI: 10.1210/endo.139.4.5879
Pelletier J, Bodin L, Hanocq E, Malpaux B, Teyssier J, Thimonier J, Chemineau P: Association between expression of reproductive seasonality and alleles of the gene for mel 1a receptor in the ewe. Biol Reprod, 62, 1096-1101, 2000. DOI: 10.1095/biolreprod62.4.1096
Muselin F, Dumitrescu E, Cristina R, Doma A, Trif A: Protective effect of melatonin on aluminum accumulation in some organs of rats. J Fac Vet Med Istanbul Univ, 41, 26-30, 2015. DOI: 10.16988/iuvfd.2015.88209
Martinez-Royo A, Lahoz B, Alabart JL, Folch J, Calvo JH: Characterisation of the melatonin receptor 1A (MTNR1A) gene in the Rasa Aragonesa sheep breed: Association with reproductive seasonality. Anim Reprod Sci, 133, 169-175, 2012. DOI: 10.1016/j.anireprosci.2012.06.018
Mateescu RG, Lunsford AK, Thonney ML: Association between melatonin receptor 1A gene polymorphism and reproductive performance in Dorset ewes. J Anim Sci, 87, 2485-2488, 2009. DOI: 10.2527/jas.2008
Reppert SM, Weaver DR, Ebisawa T: Cloning and characterization of a mammalian melatonin receptor that mediates reproductive and circadian responses. Neuron, 13, 1177-1185, 1994. DOI: 10.1016/0896- (94)90055-8
Carcangiu V, Mura MC, Vacca GM, Dettori ML, Pazzola M, Daga C, Luridiana S: Characterization of the melatonin receptor gene MT1 in Mouflon (Ovis Gmelini Musimon) and its relationship with reproductive activity. Mol Reprod Dev, 77, 196, 2010. DOI: 10.1002/mrd.21125
Wright CW: Polymorphisms at the melatonin (MTNR1A) gene and their association to reproductive performance in fall lambing ewes. MS Thesis, South Dakota State Univ, 2000.
Teyssier J, Migaud M, Debus N, Maton C, Tillard E, Malpaux B, Chemineau P, Bodin L: Expression of seasonality in Merinos d'Arles ewes of different genotypes at the MT1 melatonin receptor gene. Animal, 5, 36, 2011. DOI: 10.1017/S1751731110001813
Chu MX, Cheng DX, Liu WZ, Fang L, Ye SC: Association between melatonin receptor 1A gene and expression of reproductive seasonality in sheep. Asian-Aust J Anim Sci, 19, 1079-1084, 2006. DOI: 10.5713/ ajas.2006.1079
Hernandez X, Bodin L, Chesneau D, Guillaume D, Chemineau P, Malpaux B, Migaud M: Relationship between MT1 melatonin receptor gene polymorphism and seasonal physiological responses in Île-de-France ewes. Reprod Nutr Dev, 45, 151-162, 2005. DOI: 10.1051/ rnd:2005012
Kaczor U, Kmiecik M, Molik E, Rychlik T: Polymorphism in the melatonin receptor gene MT1 (locus MTNR1A) in sheep. Arch Tierz, 49, 262, 2006.
Shahroudi FE, Nassiry MR, Valizadh R, Heravi A: Genetic poly- morphism at MTNR1A, CAST and CAPN loci in Iranian Karakul sheep. Iran J Biotech, 4, 117-122, 2006.
Faigl V, Arnyasi M, Keresztes M, Kulcsar M, Reiczigel J, Danko G, Javor A, Cseh S, Huszeniccza G: Seasonality of reproduction and MT1 receptor gene polymorphism in Awassi sheep. Reprod Domest Anim , 11, 2008. DOI: 10.1111/j.1439-0531.2008.01232.x
Şeker İ, Özmen Ö, Çınar Kul B, Ertuğrul O: Polymorphism in melatonin receptor 1A (MTRN1A) gene in Chios, White Karaman and Awassi sheep breeds. Kafkas Univ Vet Fak Derg, 17, 865-868, 2011. DOI: 9775/kvfd.2010.3811
Elmacı C, Şahin Ş, Öner Y: Distribution of different alleles of aromatase cytochrome P450 (CYP19) and melatonin receptor 1A (MTRN1A) genes among native Turkish sheep breeds. Kafkas Univ Vet Fak Derg, 19, 929-933, DOI: 10.9775/kvfd.2013.8900
Carcangiu V, Mura MC, Vacca GM, Pazzola M, Dettori ML, Luridiana S, Bini PP: Polymorphism of the Melatonin Receptor MT1 gene and its relationship with seasonal reproductive activity in the Sarda sheep breed. Anim Rep Sci, 116, 65-72, 2009. DOI: 10.1016/j.anireprosci.2009.01.005
Notter DR, Cockett NE, Hadfield TS: Evaluation of melatonin receptor 1A as a candidate gene influencing reproduction in an autumn- lambing sheep flock. J Anim Sci, 81, 912-917, 2003.
Hristova D, Georgieva S, Yablanski T, Tanchev S, Slavov R, Bonev G: Genetic polymorphism of the melatonin receptor MT1 gene in four Bulgarian sheep breeds. Agri Sci Tech, 4, 187-192, 2012.
Hatami M, Rahimi Mianji G, Farhadi A: Association of melatonin receptor 1A gene polymorphisms with production and reproduction traits in Zandi sheep. Iran J App Anim Sci 4, 75-78, 2014.
Moradi N, Rahimi Mianji G, Nazifi N, Nourbakhsh A: Polymorphism of the melatonin receptor 1A gene and its association with litter size in Zel and Naeini sheep breeds. Iran J Appl Anim Sci, 4, 79-87, 2014.
Saxena VK, Jha BK, Meena AS, Naqvi SMK: Sequence analysis and identification of new variations in the coding sequence of melatonin receptor gene (MTNR1A) of Indian Chokla sheep breed. Meta Gene, 2, 450- , 2014. DOI: 10.1016/j.mgene.2014.05.005
Saxena VK, Jha BK, Meena AS, Narula HK, Kumar D, Naqvi SMK: Assessment of genetic variability in the coding sequence of melatonin receptor gene (MTNR1A) in Tropical Arid sheep breeds of India. Reprod Domestic Anim, 50, 517-521, 2015. DOI: 10.1111/rda.12503
Trecherel E, Batailler M, Chesneau D, Delagrange P, Malpaux B, Chemineau P, Migaud M: Functional characterization of polymorphic variants for ovine MT1 melatonin receptors: Possible implication for seasonal reproduction in sheep. Anim Reprod Sci, 122, 328-334, 2010. DOI: 10.1016/j.anireprosci.2010.10.007
Messer LA, Wang L, Tuggle CK, Yerle M, Chardon P, Pomp D, Womack JE, Barendse W, Crawford AM, Notter DR, Rothschild MF: Mapping of the melatonin receptor la (MTNR1A) gene in pigs, sheep, and cattle. Mamm Genome, 8, 368-370, 1997. DOI: 10.1007/s003359900444
Yeh F, Yang RC, Boyle T: Popgene (v.1.32) Microsoft Windows-based freeware for Population Genetic Analysis, 2000. http://www.ualberta. ca/~fyeh/Pop32.exe; Accessed: 04.03.2015.
Sezenler T, Koycu E, Yaman Y, Ceyhan A, Küçükkebapçı M, Yüksel MA: Determining some reproductive characteristics of Kıvırcık, Gökçeada and Sakız sheep races. TAGEM/95K120250 Project Report, Bandırma,