Imaging pancreatic beta-cells in the intact pancreas.
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.
Hara
Manami
M
Dept. of Medicine, University of Chicago, 5841 South Maryland Ave., MC1027, Chicago, IL 60637, USA. mhara@midway.uchicago.edu
Dizon
Restituto F
RF
Glick
Benjamin S
BS
Lee
Catherine S
CS
Kaestner
Klaus H
KH
Piston
David W
DW
Bindokas
Vytautas P
VP
D000818:Animals; D000831:Animals, Newborn; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D001652:Bile Ducts / Q000166:cytology; D001808:Blood Vessels / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007328:Insulin / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000166:cytology*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence / Q000379:methods; D009419:Nerve Tissue Proteins / Q000235:genetics; D010179:Pancreas / Q000166:cytology* / Q000196:embryology; D010183:Pancreatic Ducts / Q000166:cytology; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
D051792:Basic Helix-Loop-Helix Proteins; D049452:Green Fluorescent Proteins; D007328:Insulin; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C485182:Cyan Fluorescent Protein; C407737:Neurog3 protein, mouse