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Am J Physiol Endocrinol Metab 290: E1041-E1047, 2006. First published December 20, 2005; doi:10.1152/ajpendo.00365.2005
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INNOVATIVE METHODOLOGY

Imaging pancreatic beta-cells in the intact pancreas

Manami Hara,1 Restituto F. Dizon,1 Benjamin S. Glick,2 Catherine S. Lee,4 Klaus H. Kaestner,4 David W. Piston,5 and Vytautas P. Bindokas3

Departments of 1Medicine, 2Molecular Genetics and Cell Biology, and 3Neurobiology, Pharmacology and Physiology, University of Chicago, Chicago, Illinois; 4Department of Genetics, University of Pennsylvania, Philadelphia, Pennsylvania; and 5Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, Tennessee

Submitted 4 August 2005 ; accepted in final form 12 December 2005

We have developed a method to visualize fluorescent protein-labeled beta-cells in the intact pancreas through combined reflection and confocal imaging. This method provides a 3-D view of the beta-cells in situ. Imaging of the pancreas from mouse insulin I promoter (MIP)-green (GFP) and red fluorescent protein (RFP) transgenic mice shows that islets, beta-cell clusters, and single beta-cells are not evenly distributed but are aligned along the large blood vessels. We also observe the solitary beta-cells in both fetal and adult mice and along the pancreatic and common bile ducts. We have imaged the developing endocrine cells in the embryos using neurogenin-3 (Ngn3)-GFP mice crossed with MIP-RFP mice. The dual-color-coded pancreas from embryos (E15.5) shows a large number of green Ngn3-expressing proendocrine cells with a smaller number of red beta-cells. The imaging technique that we have developed, coupled with the transgenic mice in which beta-cells and beta-cell progenitors are labeled with different fluorescent proteins, will be useful for studying pancreatic development and function in normal and disease states.

development of pancreas; islet formation


Address for reprint requests and other correspondence: M. Hara, Dept. of Medicine, University of Chicago, 5841 South Maryland Ave., MC1027, Chicago, IL 60637 (e-mail: mhara{at}midway.uchicago.edu)






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