Kenez_2014_J.Dairy.Sci_97_3626

A balanced lipolytic regulation in adipose tissues based on fine-tuning of prolipolytic and antilipolytic pathways is of vital importance to maintain the metabolic health in dairy cows. Antilipolytic pathways, such as the G protein-coupled receptor 109A (GPR109A)-mediated pathway and the insulin signaling pathway in bovine adipose tissues may be involved in prohibiting excessive lipomobilization by reducing triglycerol hydrolysis. This study aimed to evaluate the in vitro antilipolytic potential of the mentioned pathways in bovine adipose tissue explants. Therefore, subcutaneous and retroperitoneal adipose tissue samples (approximately 100mg) of German Holstein cows were treated for 90min ex vivo with nicotinic acid (2, 8, or 32muM), nicotinamide (2, 8, or 32muM), beta-hydroxybutyrate (0.2, 1, or 5mM), or insulin (12mU/L), with a concurrent lipolytic challenge provoked with 1muM isoproterenol. Lipolytic and antilipolytic responses of the adipose tissues were assessed by measuring free glycerol and nonesterified fatty acid release. To identify molecular components of the investigated antilipolytic pathways, protein abundance of GPR109A and the extent of hormone-sensitive lipase (HSL) phosphorylation at serine residue 563 were detected by Western blotting. Treatment with nicotinic acid or beta-hydroxybutyrate decreased the lipolytic response in adipose tissue explants and concurrently reduced the extent of HSL phosphorylation, but treatment with nicotinamide or insulin did not. Subcutaneous adipose tissue constitutively expressed more GPR109A protein, but no other depot-specific differences were observed. This study provides evidence that the GPR109A-mediated pathway is functionally existent in bovine adipose tissues, and confirms that HSL phosphorylation at serine residue 563 is also important in antilipolytic regulation in vitro. This antilipolytic pathway may be involved in a balanced lipid mobilization in the dairy cow.