PanIN-associated pericyte, glial, and islet remodeling in mice revealed by 3D pancreatic duct lesion histology

Lin, Pei Yu; Peng, Shih-Jung; Shen, Chia‐Ning; Pasricha, Pankaj J.; Tang, Shiue–Cheng

doi:10.1152/ajpgi.00071.2016

Cited by 21 publications

(24 citation statements)

References 43 publications

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“…The technical advance of tissue clearing of mouse and human gastrointestinal tissues [10,37,[41][42][43] has enabled us to develop 3D histology for visualisation of the pancreatic neuroinsular network. Using the transparent specimens, we applied both fluorescence and transmitted light microscopy (a dualoptical approach) to examine the pancreatic microstructure and neurovascular network.…”

Section: Discussionmentioning

confidence: 99%

Human pancreatic neuro-insular network in health and fatty infiltration

et al. 2017

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Aims/hypothesis Identification of a pancreatic neuro-insular network in mice suggests that a similar integration of islets and nerves may be present in the human pancreas. To characterise the neuro-insular network and the intra-pancreatic ganglia in a clinically related setting, we examined human pancreases in health and with fatty infiltration via 3-dimensional (3D) histology and compared the human pancreatic microenvironment with its counterpart in mice. Methods Human pancreatic specimens from individuals with normal BMI, high BMI (≥ 25) and type 2 diabetes were used to investigate the neuro-insular network. Transparent specimens were prepared by tissue clearing for transmitted light and deeptissue fluorescence imaging to simultaneously visualise infiltrated adipocytes, islets and neurovascular networks. Results High-definition images of human islets reveal that both the sympathetic and parasympathetic nerves enter the islet core and reside in the immediate microenvironment of islet cells. Around the islets, the neuro-insular network is visualised with 3D histology to identify the intra-pancreatic ganglia (peri-lobular and intra-parenchymal ganglia) and the islet-ganglionic association. In humans, but not in mice, pancreatic fatty infiltration (BMI dependent) features adipocytes infiltrating into the parenchyma and accumulating in the perilobular space, in which the peri-lobular ganglia also reside. We identified the formation of adipose-ganglionic complexes in the peri-lobular space and enlargement of ganglia around adipocytes. In the specimen from the individual with type 2 diabetes, an increase in the number of nerve projections from the intra-parenchymal ganglia is associated with severe fatty infiltration. Conclusions/interpretation We present new perspectives of human pancreas and islet innervation via 3D histology. Our results strongly suggest that fatty infiltration in the human pancreas creates a neurotrophic microenvironment and promotes remodelling of pancreatic innervation.

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Section: Discussionmentioning

confidence: 99%

Human pancreatic neuro-insular network in health and fatty infiltration

et al. 2017

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“…Importantly, with the addition of multicolor immunolabeling, the 3D relationships of normal ducts, precursor lesions, invasive carcinoma, immune cells, islets of Langerhans, vessels, and nerves should be visualizable. 10,15 Clearing methodologies have been described for 100 years. 7 The more recent addition of immunolabeling using fluorescent antibodies has opened the door to 3D microscopy.…”

Section: Discussionmentioning

confidence: 99%

“…When DNA and RNA studies are added in the future, the methodology may be used to define the drivers of 3D morphology. 10,11…”

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confidence: 99%

Immunolabeling of Cleared Human Pancreata Provides Insights into Three-Dimensional Pancreatic Anatomy and Pathology

Noë

Rezaee

Asrani

et al. 2018

The American Journal of Pathology

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Visualizing pathologies in three dimensions can provide unique insights into the biology of human diseases. A rapid and easy-to-implement dibenzyl ether-based technique was used to clear thick sections of surgically resected human pancreatic parenchyma. Protocols were applicable to both fresh and formalin-fixed, paraffin-embedded tissue. The penetration of antibodies into dense pancreatic parenchyma was optimized using both gradually increasing antibody concentrations and centrifugal flow. Immunolabeling with antibodies against cytokeratin 19 was visualized using both light sheet and confocal laser scanning microscopy. The technique was applied successfully to 26 sections of pancreas, providing three-dimensional (3D) images of normal pancreatic tissue, pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasms, and infiltrating pancreatic ductal adenocarcinomas. 3D visualization highlighted processes that are hard to conceptualize in two dimensions, such as invasive carcinoma growing into what appeared to be pre-existing pancreatic ducts and within venules, and the tracking of long cords of neoplastic cells parallel to blood vessels. Expanding this technique to formalin-fixed, paraffin-embedded tissue opens pathology archives to 3D visualization of unique biosamples and rare diseases. The application of immunolabeling and clearing to human pancreatic parenchyma provides detailed visualization of normal pancreatic anatomy, and can be used to characterize the 3D architecture of diseases including pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasm, and pancreatic ductal adenocarcinomas.

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“…Propidium iodide staining (50 μg/ml, Invitrogen) was performed to reveal the nuclei. The labelled specimens were immersed in the optical clearing aqueous solution (RapiClear 1.52 solution [18,19], SunJin Lab, Hsinchu, Taiwan) 1 day before being imaged via confocal microscopy.…”

Section: Methodsmentioning

confidence: 99%