Abstract
Proper identification of pancreatic ducts is a major challenge for researchers performing partial duct ligation (PDL), because pancreatic ducts, which are covered with acinar cells, are translucent and thin. Although damage to pancreatic ducts may activate quiescent ductal stem cells, which may allow further investigation into ductal stem cells for therapeutic use, there is a lack of effective techniques to visualize pancreatic ducts. In this study, we report a new method for identifying pancreatic ducts. First, we aimed to visualize pancreatic ducts using black, waterproof fountain pen ink. We injected the ink into pancreatic ducts through the bile duct. The flow of ink was observed in pancreatic ducts, revealing their precise architecture. Next, to visualize pancreatic ducts in live animals, we injected fluorescein-labeled bile acid, cholyl-lysyl-fluorescein into the mouse tail vein. The fluorescent probe clearly marked not only the bile duct but also pancreatic ducts when observed with a fluorescent microscope. To confirm whether the pancreatic duct labeling was successful, we performed PDL on Neurogenin3 (Ngn3)-GFP transgenic mice. As a result, acinar tissue is lost. PDL tail pancreas becomes translucent almost completely devoid of acinar cells. Furthermore, strong activation of Ngn3 expression was observed in the ligated part of the adult mouse pancreas at 7 days after PDL.
Highlights
More than 420 million people suffer from diabetes worldwide.[1]
The ink injection strategy allowed us to identify pancreatic ducts, we screened for chemicals that are not harmful to the animal, so that it could be used in live animals
CLF labeled pancreatic ducts (Fig. 2B, C). Both head and tail pancreatic ducts were clearly visualized in live animals. To fully understand this method for visualizing pancreatic ducts, we investigated whether this method affects any other organs and tissues
Summary
More than 420 million people suffer from diabetes worldwide.[1]. Slowing the increasing prevalence of diabetes is a major challenge in the medical field, because no effective drug presently exists. Transplantation of pancreatic islets is a reliable diabetes treatment that is presently available.[2,3] Islets can be generated from pancreatic progenitor cells that emerge when the tissue is damaged Where these progenitor cells reside remains controversial.[4,5,6,7,8] It is generally thought that beta cell growth occurs only by self-renewal of mature beta cells under normal physiological conditions.[9,10,11] In other studies, it has been reported that beta cell progenitors can be generated by partial duct ligation (PDL) in adult mice.[12,13,14] The procedure involves ligation of the pancreatic duct that drains pancreatic enzymes into the duodenum. The outcomes of studies using the ligation technique appear to vary across
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