Meeting Abstract

P2.15  Tuesday, Jan. 5  Using Nkx6.1 to detect pancreatic beta-cells in the American alligator FREYMILLER, HJ*; HARTY, JJ; EDWARDS, TM; University of Florida; University of Florida; Tulane University haley87@ufl.edu

The antibody F55A10 detects the pancreatic transcription factor Nkx6.1 , which is specific to insulin-producing beta cells, and thus provides a method of selective staining. Nkx6.1 has been used to mark beta cells in both mouse, rat, pig, zebrafish, human and chicken . Being that chicken and alligators are evolutionarily very close, we hypothesized that this antibody would prove to be a successful marker for Alligator mississippiensis beta cells. Prior to staining, peroxidase activity was blocked using hydrogen peroxide. Following this, the primary antibody, Nkx.6.1 was applied overnight at 4⁰C. A panspecific biotinylated secondary antibody bridge, followed by a streptavidin/peroxidase complex was used to link the antibody to the 3,3-diaminobenzene (DAB) dye, which allows for visualization. Nkx6.1 was a successful marker in 5 month old female alligator pancreas. Alligator beta cells are scattered as single cells, in small clusters, and in asymmetrical islets in the pancreas. This stain can be used to assess beta cell quantity in later studies.