Imaging pancreatic beta-cells in the intact pancreas

• Published in: American journal of physiology: endocrinology and metabolism 2006-05, Vol.290 (5), p.E1041-E1047
• Main Authors: Hara, Manami, Dizon, Restituto F, Glick, Benjamin S, Lee, Catherine S, Kaestner, Klaus H, Piston, David W, Bindokas, Vytautas P
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Basic Helix-Loop-Helix Transcription Factors - genetics; Bile Ducts - cytology; Blood Vessels - cytology; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Image Processing, Computer-Assisted; Imaging, Three-Dimensional - methods; Insulin - genetics; Insulin-Secreting Cells - cytology; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Mice; Mice, Transgenic; Microscopy, Fluorescence - methods; Nerve Tissue Proteins - genetics; Pancreas - cytology; Pancreas - embryology; Pancreatic Ducts - cytology; Promoter Regions, Genetic - genetics; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Red Fluorescent Protein
• ISSN: 0193-1849; 1522-1555
• DOI: 10.1152/ajpendo.00365.2005

Departments of 1 Medicine, 2 Molecular Genetics and Cell Biology, and 3 Neurobiology, Pharmacology and Physiology, University of Chicago, Chicago, Illinois; 4 Department of Genetics, University of Pennsylvania, Philadelphia, Pennsylvania; and 5 Department 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 -cells in the intact pancreas through combined reflection and confocal imaging. This method provides a 3-D view of the -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, -cell clusters, and single -cells are not evenly distributed but are aligned along the large blood vessels. We also observe the solitary -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 -cells. The imaging technique that we have developed, coupled with the transgenic mice in which -cells and -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 )