Determining Beta Cell Mass, Apoptosis, Proliferation, and Individual Beta Cell Size in Pancreatic Sections
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Pancreatic beta cells have a significant remodeling capacity which plays an essential role in the maintenance of glucose homeostasis. Beta cell apoptosis, replication, size, dedifferentiation, and (neo)generation contribute to the beta cell mass regulation. However, the extent of their respective contribution varies significantly depending on the specific condition, and it is the balance among them that determines the eventual change in beta cell mass. Thus, the study of the pancreatic beta cell mass regulation requires the determination of all these factors. In this chapter, we describe the quantification of beta cell replication based on the incorporation of thymidine analogs into replicated DNA strands and on the expression of Ki67 antigen and phosphorylation of histone H3. Beta cell apoptosis is analyzed by the TUNEL technique, and beta cell mass and cross-sectional area of individual beta cells are determined by computerized image processing methods.
Key wordsBrdU EdU PHH3 Ki67 antigen Replication TUNEL Apoptosis Beta cell mass Cross-sectional individual beta cell area Pancreatic islets
This work has been supported by grants from the Catalan Diabetes Association (NT), University of Barcelona (NT), and Carlos III Health Institute (ISCIII) PI16/00462 co-funded by FEDER funds/European Regional Development Fund (ERDF) – “A Way to Build Europe” (EM) – and by CIBERDEM which is a project of ISCIII.
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