leptin and calpastatin

Single nucleotide polymorphisms (SNPs) carried in calpain (CAPN1), calpastatin (CAST), and leptin (LEP) genes are associated with meat tenderness. Due to the economic importance of this meat quality attribute, the development of fast, reliable, and affordable methods to identify bovine carriers of favorable alleles is of great importance for genetic improvement. Currently, PCR-RFLP is accepted as the standard gold method for genotyping SNPs associated with meat tenderness. But these SNPs can be detected by other techniques as high-resolution melting (HRM) analysis - a post-PCR method - that offers several advantages and has great application potential in the meat industry. In this study, we standardized, validated, and compared the performance of PCR-HRM to that of PCR-RFLP in genotyping bovine SNPs associated with meat tenderness: CAPN4751, CAPN316, CAST2959, CAST282, LEPE2FB, and LEPE2JW. We analyzed genotypes of a total of 380 bovines, 110 Bos taurus and 270 Bos indicus. Results obtained with PCR-HRM were consistent with those found by PCR-RLFP. Furthermore, HRM was found to be highly sensitive, and our results confirmed the repeatability (intra-assay precision) and reproducibility (inter-assay precision) of this assay. An internal control for endonuclease activity was created using site-directed mutagenesis to generate an additional enzymatic restriction point useful to discriminate SNP alleles. Our results show that PCR-HRM is an efficient method that produces reliable and rapid results. However, should be had in account that the method of DNA extraction, the quality and quantity of DNA, analyst-related variations, and primer design may generate challenges for allele discrimination.

Topics: Animals; Calcium-Binding Proteins; Calpain; Cattle; Genotyping Techniques; Leptin; Meat; Polymerase Chain Reaction; Polymorphism, Single Nucleotide

The aim of this study was to assess the associations between the single nucleotide polymorphisms in CAPN1, CAST, DGAT1, FABP4, LEP, RORC and SCD1 genes and the sensory meat quality in an intensively fed commercial population (Charolais, Limousin and Retinta breed). This work carried out analyses on a common Spanish population and evaluated the association between the markers and sensory traits. A total of 161 bulls were allocated to two different feedlots with two different finishing diets. Steaks aged for 7 and 21 days were assessed by both untrained and trained sensory panels. A significant association and allelic substitution effect were observed for markers UoG-CAST, LEP: g.73C>T and SCD1: g.878T>C on different descriptors evaluated by a consumer panel (tenderness and overall acceptability). There are no precedents of these kinds of association studies in a Spanish commercial population. The study suggested that CAST, LEP and SCD1 genes have a potential effect on the different measurements of sensory meat quality.

Topics: Adolescent; Adult; Aged; Alleles; Animal Husbandry; Animals; Breeding; Calcium-Binding Proteins; Cattle; Female; Genotype; Humans; Leptin; Male; Middle Aged; Phenotype; Polymorphism, Single Nucleotide; Red Meat; Spain; Stearoyl-CoA Desaturase; Stress, Mechanical; Taste; Young Adult

This study aimed to investigate differential allele expression (DAE) and polymorphism and parent-of-origin effects on expression of genes related to beef traits. CAST, related to meat tenderness, and DGAT1 and leptin, related to fat deposition, were evaluated. In bovine fetal tissues CAST was expressed twice as much (P < 0.05) in muscle of homozygous GG than in heterozygous AG. Leptin was expressed about one-tenth as much (P < 0.05) in heterozygous TpCm (allele T of paternal origin and allele C of maternal origin) than in homozygous CC. No DAE was observed. The evidence of polymorphism effect on expression of CAST and parent-of-origin effect on leptin contributes to a better understanding of events controlling the expression of genes of economic interest in cattle. Furthermore, if the parent-of-origin effects observed in fetal tissues are confirmed in adult tissues and associated to phenotypic variation, this parental origin criterion may be considered in marker-assisted selection of beef traits.

Topics: Alleles; Animals; Body Composition; Calcium-Binding Proteins; Cattle; Diacylglycerol O-Acyltransferase; Gene Expression Regulation; Heterozygote; Homozygote; Inheritance Patterns; Leptin; Meat; Polymorphism, Single Nucleotide