Real-time RT-PCR and cDNA macroarray to study the impact of the genetic polymorphism at the alphas1-casein locus on the expression of genes in the goat mammary gland during lactation

Abstract

Milk fat has a large effect on nutritional, technological and sensorial properties of milk products. The milk fat content and composition are modulated by genetics and nutritional factors and imply a large number of enzymes. The regulation of their gene expression in the mammary gland still needs to be clarified. An association between the extensive polymorphism at the alphas1-casein (alphas1-Cas) locus and both the lipid content and the characteristics of this fraction in caprine milk has been demonstrated. In order to decipher the mechanism responsible for this impact, a quantification of the transcripts of four lipogenic key enzymes (acetyl-CoA carboxylase, fatty acid synthase, lipoprotein lipase and stearoyl-CoA desaturase) was performed using real-time RT-PCR, suggesting an absence of association between the alphas1-Cas genotype and expression variability of the studied genes. This approach has been completed by a more global analysis using a first generation of ruminant macroarray gathering 400 gene probes. The comparison of the expression profiles of lactating goat alphas1-Cas A/A (strong allele) and F/F (defective allele) mammary gland allowed to confirm the expected variability in the expression of known genes (such as those encoding the alphas1-casein) in ruminant mammary tissues as well as to identify up- and down-regulated genes. A second generation of ruminant cDNA macroarray extended to a few thousands of genes is currently in progress and will be applied to study different factors such as the nutritional regulation of gene expression in the mammary gland.