عنوان مقاله [English]
Introduction: Generally, genetic markers, associated with metabolic pathways, might be used in marker-assisted selection to improve production and reproduction traits in farm animals. Especially, some traits with low heritabilities, such as reproduction and health traits could not be easily improved by classic selection strategies. However, the use of genetic markers and marker-assisted selection are considered powerful tools for the genetic improvement of low-heritable traits. Thus, detection of genetic markers associated with production and reproduction traits is an effective way to save endangered indigenous breeds, such as the Markhoz goat, a mohair-producing breed in Iran. The thyroid hormone-responsive (THRSP) gene is a candidate gene involved in thyroid hormone functions and the lipogenesis process. This gene is a protein-coding gene, with two exons, located on chromosome 29 in goats. There are some reports on the association of THRSP with production traits in farm animals. There are limited studies on metabolic pathways of THRSP and its associations with important economic traits in small ruminants and no study on the association of THRSP and reproduction traits was found in the literature. The aim of the present study was the investigation of the THRSP gene polymorphism and its association with body weight and litter size traits in Markhoz goats.
Materials and methods: A total of 140 blood samples of Markhoz goats were randomly collected from the Research and Breeding Station in Kurdistan province in western Iran. Genomic DNA was extracted from whole blood samples. A pair of primers were designed using the Primer 3 online software to amplify a 486 bp fragment in THRSP gene exon 1. The designed primers were as follows:
Polymorphism of the amplified fragment was investigated using the single-strand conformational polymorphism (SSCP) method and sequencing of three random samples for each SSCP pattern. The observed sequences were aligned to the GenBank reference sequence using the MegAlign module of DNAStar software and compared based on the Clustal W method. The sequences were translated by the Translate section of the ExPASy website (us.expasy.org/translatetool). Associations of body weight traits, including birth weight, three-month, six-month, nine-month, and 12-month body weights, with the observed genotypes were investigated using a general linear model, fitting genotype, birth year, birth type, and dam age as the fixed factors. The association of the observed genotypes with litter size was investigated using a two-way Chi-squared test. The SAS 9.4 program was employed for the association analyses.
Results and discussion: In the studied samples, two different SSCP patterns and two single-nucleotide polymorphisms (SNPs) were detected in 148 and 173 bp locations of the studied fragment, as g.148G>A (resulting in Arginine to Glutamine amino acids exchange) and g.173A>G (a synonymous mutation), both as heterozygous loci. In the studied population, the frequency of the double-homozygous genotype, GG/AA (0.743), was noticeably higher than the double-heterozygous genotype, which is GA/AG (0.257). The alleles G and A had high frequencies, both equal to 0.871 in the 148 and 173 bp loci, respectively. Both loci had a significant deviation from Hardy-Weinburg equilibrium (P<0.0001). Polymorphism of the studied fragment did not have any significant effect on body weight traits. A significant association was observed between the detected genotypes and litter size. Whereby, litter size in double-heterozygous does (1.21) was significantly higher than the double-homozygous individuals (1.04), (P = 0.015). The present study is probably the first report on the association of the THRSP gene with litter size in goats. The THRSP is a protein which involves in thyroid hormone function and therefore might affect many metabolic pathways. However, a significant association of the observed genotypes with prolificacy is probably due to the role of THRSP in lipids metabolism and the association of lipids metabolism with reproduction performance.
Conclusions: The THRSP exon 1 is polymorphic in Markhoz goats. The studied fragment did not have any significant associations with body weight traits, but it had a significant effect on litter size. Based on the results of this study, the THRSP gene could be considered a candidate gene for litter size, and the detected SNPs at 148 and 173 bp of the studied fragment, could be used for marker-assisted selection in Markhoz goats. However, more studies on possible associations between THRSP polymorphism and production and reproduction traits are still needed in other goat breeds.