Detection of SGLT1 by western blot in birds

FUNES SC; GATICA SOSA CP; CHEDIACK, JG; CAVIEDES-VIDAL E

Mendoza

Congreso; XXV Reunión Científica Anual de la Sociedad de Biología de Cuyo.; 2008

Universidad Nacional de Cuyo

SGLT1, the Na+/Glucose cotransporter is a relevant component of the glucose uptake of the brush border membrane of the small intestine, though up to date no efforts had been performed to evaluate its expression in birds. Thus, our aim was to develop a western blot assay to identify SGLT1 in birds. We used two bird species Gallus gallus (n=3) and Taeniopygia guttata (n=3). We prepared jejunal homogenates of the small intestines and separated the proteins using SDS-PAGE (8%), transferred to a PVDF membranes. Western blot assays using a SGLT-1 primary antibody (Chemicon, Inc.) against amino acids 402-420 of the putative extracellular loop of SGLT1 of rat small intestine were completed. We assayed rat small intestine homogenates of Sprague-Dawley rats as well, to validate the specificity of the antibody. Immunoblotting was developed by an enhanced chemiluminescence system. Characteristic bands (~50; ~70 and ~225 kDa) of SGLT1 in rats were apparent in both bird species showing that the protein has highly conserved sequences among mammals and birds, as has been proposed for other taxa by colleagues. To our knowledge this is the first western blot assay developed for birds and prove to be a useful tool for avian digestive physiology. Supported by FONCYT 25561 and UNSL 22Q751 to ECV.