Polymorphisms in candidate genes for athletic performance and quantification of MCT1 and CD147 in red blood cells of arabian and quarter horses

Author(s)

Inaê Cristina Regatieri

• Document type: Doctoral Thesis
• Press: Jaboticabal. 2016-10-31.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Ciências Agrárias e Veterinárias. Jaboticabal
• Defense date: 2016-10-19
• Advisor: Antonio de Queiroz Neto; Guilherme de Camargo Ferraz; Rogério Abdallah Curi

Abstract

Monocarboxylate transporter isoform 1 (MCT1), present in the red blood cell membranes and its ancillary protein CD147 have as function transport H+ and lactate ions from the plasma into the red blood cells, thereby maintaining acid/base homeostasis and retarding systemic acidosis and muscular fatigue. Thereby, the aim of this study was to compare the amount of MCT1 and CD147 proteins in the red blood cells of Arabian and Quarter Horses with different levels of athletic ability. Furthermore, we investigated polymorphisms for MCT1, CD147, DMRT3, and PDK4 genes in Arabian and Quarter Horses in order to check associations between the polymorphisms and the performance in these breeds. Arabian horses were divided into two groups according to their performance in endurance competition and Quarter Horses were separated by its performance in races, determined by Speed Index. The amount of MCT1 and CD147 proteins in the plasma membrane of red blood cells was determined by western blotting analysis with arbitrary optical density units (OD), using a human specific anti-MCT1 and anti- CD147 antibody. Data for the amounts of proteins were analyzed using the PROC MIXED procedure of SAS software. The model for the analysis included the effects of sex, breed and performance group within breed as fixed effect and age as covariate. The correlations were analyzed by Pearson correlation test using the PROC CORR procedure of SAS software. P values <0.01 were considered statistically significant. Polymorphisms of the genes were analyzed by sequencing (MCT1 and CD147), PCR-RFLP (DMRT3) and ARMS-PCR (PDK4) techniques. The statistical packages Genetics, Lattice and GenABEL were used to compare the frequencies of the groups using the software R, with the Fisher's exact test being performed with significance level of 5%. MCT1 and CD147 proteins were found in the red blood cell membranes of all studied animals. The amount of MCT1 was significantly (p<0.0001) higher in Quarter Horses (2.99 ± 0.35 OD) than in Arabians (1.04 ± 0.08 OD). Quarter Horses (3.23 ± 0.38 OD) also showed bigger contents of CD147 than Arabians (0.88 ± 0.06 OD). There was not statistical difference in the amounts of MCT1 and CD147 between the performance groups of both breeds. Positive correlation was found between the amounts of MCT1 and CD147 (r=0.95; p<0.0001). The A allele from the polymorphisms Lys457Gln:1573A>C of MCT1 and Ile51Val:168A>G of CD147 gene, were fixed in both breeds. A new polymorphism (AY457175.1:c1498G>A) was found in the MCT1 gene sequence. For DMRT3 mutation, all the animals shown to have the C allele fixed for the polymorphism. Arabians showed significant greater frequency of the G allele than Quarter Horses (p<0.01) for the PDK4 polymorphism. However, there was not difference between the groups of performance for both breeds. In summary, it follows that the Quarter Horses have greater amount of MCT1 and CD147 proteins than Arabian. It was not possible to determine the influence of polymorphisms in MCT1, CD147 and DMRT3 genes in the athletic performance of these breeds since they had alleles fixed. There was a significant difference in the frequencies of the PDK4 polymorphism between Arabians and Quarter Horses, but there was not difference between the performance groups. (AU)

• FAPESP's process: 12/24193-0 - POLYMORPHISMS AND EXPRESSION OF MONOCARBOXYLATE TRANSPORTER MCT1 AND THE ANCILLARY PROTEIN CD147 IN RED BLOOD CELLS IN ARABIAN AND QUARTER HORSES
• Grantee: Inaê Cristina Regatieri
• Support Opportunities: Scholarships in Brazil - Doctorate