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Leading researchers and clinical investigators describe their best cutting-edge techniques for studying, at both the molecular and biochemical levels, the defects in insulin production and action associated with diabetes. Written in step-by-step detail to ensure ready reproducibility and robust results, these techniques allow investigators, both novices and those already active in the field, to study every major facet of insulin production and action. Each protocol includes an introduction to the technique, an explanation of its application, and a list of materials. Practical notes discuss how to avoid pitfalls, as well as how to adapt the methods to your own research.
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M E T H O D S I N M O L E C U L A R M E D I C I N E TM Diabetes Mellitus Methods and Protocols Edited by by Edited Sabire Özcan Humana Press 1 Isolation of Islets of Langerhans from Rodent Pancreas Colleen B. Kelly, Libby A. Blair, John A. Corbett, and Anna L. Scarim 1. Introduction Pancreatic b-cells, responsible for the synthesis and secretion of insulin in response to a glucose challenge, are located in the islets of Langerhans. Islets are comprised of a heterogeneous population of endocrine cells, including insulin-producing b-cells (approx. 65–70%), glucagon-secreting a-cells (20–25%), somatostatin-secreting d-cells, and polypeptide (PP)-secreting cells. Much of the cellular and biochemical information concerning the mechanisms by which glucose stimulates insulin secretion by pancreatic b-cells has been obtained in studies using islets isolated from rodents (1). Rat islets provide an ideal source of insulin-producing tissue to study pancreatic b-cell function as insulin secretion by rat islets closely parallels insulin secretion by human islets and it is possible to obtain a large number of islets (300–600) from a single rat pancreas. With the widespread