Dynamics of embryonic pancreas development using real-time imaging.
Current knowledge about developmental processes in complex organisms has relied almost exclusively on analyses of fixed specimens. However, organ growth is highly dynamic, and visualization of such dynamic processes, e.g., real-time tracking of cell movement and tissue morphogenesis, is becoming increasingly important. Here, we use live imaging to investigate expansion of the embryonic pancreatic epithelium in mouse. Using time-lapse imaging of tissue explants in culture, fluorescently labeled pancreatic epithelium was found to undergo significant expansion accompanied by branching. Quantification of the real-time imaging data revealed lateral branching as the predominant mode of morphogenesis during epithelial expansion. Live imaging also allowed documentation of dynamic beta-cell formation and migration. During in vitro growth, appearance of newly formed beta-cells was visualized using pancreatic explants from MIP-GFP transgenic animals. Migration and clustering of beta-cells were recorded for the first time using live imaging. Total beta-cell mass and concordant aggregation increased during the time of imaging, demonstrating that cells were clustering to form "pre-islets". Finally, inhibition of Hedgehog signaling in explant cultures led to a dramatic increase in total beta-cell mass, demonstrating application of the system in investigating roles of critical embryonic signaling pathways in pancreas development including beta-cell expansion. Thus, pancreas growth in vitro can be documented by live imaging, allowing visualization of the developing pancreas in real-time.
Puri
Sapna
S
Diabetes Center, Department of Medicine, University of California, San Francisco, CA 94143, USA.
Hebrok
Matthias
M
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002465:Cell Movement; D046149:Embryo Culture Techniques; D005260:Female; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D053823:Hedgehog Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology; D018398:Homeodomain Proteins / Q000235:genetics; D007328:Insulin / Q000032:analysis; D050417:Insulin-Secreting Cells / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D009024:Morphogenesis; D038081:Organogenesis; D010179:Pancreas / Q000737:chemistry / Q000166:cytology / Q000196:embryology*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics
D015415:Biomarkers; D053823:Hedgehog Proteins; D018398:Homeodomain Proteins; D007328:Insulin; D012333:RNA, Messenger; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D049452:Green Fluorescent Proteins