Pancreatic remodeling: beta-cell apoptosis, proliferation and neogenesis, and the measurement of beta-cell mass and of individual beta-cell size.

The endocrine pancreas has a significant remodeling capacity that plays a crucial role in the maintenance of glucose homeostasis. Changes in beta-cell apoptosis, replication and size, and islet neogenesis contribute to the remodeling of the endocrine pancreas. The extent of their respective contribution varies significantly depending on the specific condition under consideration, and it is the balance among them that determines the eventual change in beta-cell mass. Thus, the study of pancreatic remodeling requires the determination of all these factors. In this chapter, we describe the quantification of beta-cell replication based on 3-bromo-2-deoxyuridine incorporation into DNA and on the expression of Ki67 antigen, of beta-cell apoptosis by the TUNEL technique and islet neogenesis by quantification of number of islets budding from pancreatic ducts, by confocal assessment of the expression of islet cell hormones in ductal cells, and by identification of small group of extra-islet beta-cells. Point counting morphometry is used to estimate beta-cell mass and planimetry to determine the cross-sectional area of individual beta-cells, a measure of beta-cell size.