Vessel painting of the microcirculation using fluorescent lipophilic tracers

Ravnic, Dino J.; Jiang, Xian; Wolloscheck, Tanja; Pratt, Joanne H.; Huss, Harold T.; Mentzer, Steven J.; Konerding, Moritz A.

doi:10.1016/j.mvr.2005.06.002

Cited by 42 publications

(46 citation statements)

References 10 publications

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“…Tissue labelling Vessel painting [30,34,35] was performed by cardiac perfusion of the lectin (wheat germ agglutinin)-Alexa Fluor 488 conjugate (30 μg/g body weight; Cat. No.…”

Section: Methodsmentioning

confidence: 99%

“…This allowed the identification of the spatial features of the GFAP + Schwann cells and NG2 + pericytes (note: although named as a neuron-'glial' antigen, the NG2 cell-surface chondroitin sulfate proteoglycan is highly expressed in the pancreatic/islet pericytes and has been used as a histological marker [5,33]) and their association with the islet lesion and vasculature [13,17,[34][35][36]. Qualitative and quantitative analyses of the islet injury-induced gliosis and pericyte remodelling around the feeding arteriole, and their pathophysiological implications, are presented and discussed in this report.…”

Section: Introductionmentioning

confidence: 99%

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Plasticity of Schwann cells and pericytes in response to islet injury in mice

et al. 2013

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Aims/hypothesis Islet Schwann (glial) cells and pericytes are the microorgan's accessory cells positioned at the external and internal boundaries facing the exocrine pancreas and endothelium, respectively, adjacent to the endocrine cells. Plasticity of glial cells and pericytes is shown in the glial scar formation after injury to the central nervous system. It remains unclear whether similar reactive cellular responses occur in insulitis. We applied three-dimensional (3D) histology to perform qualitative and quantitative analyses of the islet Schwann cell network and pericytes in normal, streptozotocin-injected (positive control of gliosis) and NOD mouse models. Methods Vessel painting paired with immunostaining of mouse pancreatic tissue was used to reveal the islet Schwann cells and pericytes and their association with vasculature. Transparent islet specimens were prepared by optical clearing to facilitate 3D confocal microscopy for panoramic visualisation of the tissue networks.Results In-depth microscopy showed that the islet Schwann cell network extends from the peri-islet domain into the core. One week after streptozotocin injection, we observed intraislet perivascular gliosis and an increase in pericyte density. In early/moderate insulitis in the NOD mice, perilesional gliosis occurred at the front of the lymphocytic infiltration with atypical islet Schwann cell morphologies, including excessive branching and perivascular gliosis. Meanwhile, pericytes aggregated on the walls of the feeding arteriole at the peri-and intralesional domains with a marked increase in surface marker density. Conclusions/interpretationThe reactive cellular responses demonstrate plasticity and suggest a stop-gap mechanism consisting of the Schwann cells and pericytes in association with the islet lesion and vasculature when injury occurs.

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“…Tissue labelling Vessel painting [30,34,35] was performed by cardiac perfusion of the lectin (wheat germ agglutinin)-Alexa Fluor 488 conjugate (30 μg/g body weight; Cat. No.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Plasticity of Schwann cells and pericytes in response to islet injury in mice

et al. 2013

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“…Vessel painting [13,21] was performed by cardiac perfusion of the lectin (wheat germ agglutinin)-Alexa Fluor 488, 555 or 633 conjugate (30 μg/g of body weight; Invitrogen, Carlsbad, CA, USA) followed by 4% paraformaldehyde perfusion fixation. Afterwards, pancreases were harvested and the vibratome sections of the tissue (∼400 μm) were post-fixed in 4% paraformaldehyde solution for 1 h at 25°C.…”

Section: Methodsmentioning

confidence: 99%

“…use of the immersion solution to reduce scattering as light travels in the specimen [18][19][20]) to allow for deeptissue microscopy. Here, in islet imaging, we employed the same penetrative 3-D microscopy with vessel painting (perfusion of blood vessels with fluorescent probes) [13,21] and islet histology [7,8,22,23] to provide a global and integral view of the islet sympathetic neurovascular complex.…”

Section: Introductionmentioning

confidence: 99%

3-D imaging and illustration of the perfusive mouse islet sympathetic innervation and its remodelling in injury

Chiu

Hua

et al. 2012

Diabetologia

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Aims/hypothesis Sympathetic nerves influence islet hormone levels in the circulation. Insights into islet sympathetic innervation and its remodelling in diabetes may impact future therapeutics. However, standard immunohistochemistry and microtome-based microscopy cannot provide an integral view of the islet neurovascular complex. We prepared transparent islet specimens to investigate the spatial relationship between sympathetic nerves, blood vessels and islet cells in normal, streptozotocin-injected and non-obese diabetic mouse models. Methods Cardiac perfusion of fluorescent lectin was used to label pancreatic blood vessels. Tyrosine hydroxylase and nuclear staining were used to reveal islet sympathetic innervation and microstructure. Optical clearing (i.e. use of immersion solution to reduce scattering) was applied to enable 3-dimensional confocal microscopy of islets to visualise the sympathetic neurovascular complex in space. Results Unlike previously reported morphology, we observed perfusive intra-islet, perivascular sympathetic innervation, in addition to peri-islet contacts of sympathetic nerves with alpha cells and sympathetic fibres encircling the adjacent arterioles. The intra-islet axons became markedly prominent in streptozotocin-injected mice (2 weeks after injection). In non-obese diabetic mice, lymphocytic infiltration remodelled the peri-islet sympathetic axons in early insulitis. Conclusions/interpretation We have established an imaging approach to reveal the spatial features of mouse islet sympathetic innervation. The neurovascular complex and sympathetic nerve-alpha cell contact suggest that sympathetic nerves modulate islet hormone secretion through blood vessels, in addition to acting directly on alpha cells. In islet injuries, sympathetic nerves undergo different remodelling in response to different pathophysiological cues.

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“…Tissue labeling. Blood vessels of the kidney and islet graft were labeled by vessel painting (13,33) via cardiac perfusion of the lectin-Alexa Fluor 488 conjugate (30 g/g of body wt, catalog no. W11261; Invitrogen) followed by 4% paraformaldehyde perfusion fixation.…”

Section: Animals and Islet Transplantationmentioning

confidence: 99%

Three-dimensional islet graft histology: panoramic imaging of neural plasticity in sympathetic reinnervation of transplanted islets under the kidney capsule

Juang

Peng

Kuo

et al. 2014

American Journal of Physiology-Endocrinology and Metabolism

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Juang J, Peng S, Kuo C, Tang S. Three-dimensional islet graft histology: panoramic imaging of neural plasticity in sympathetic reinnervation of transplanted islets under the kidney capsule. Am J Physiol Endocrinol Metab 306: E559 -E570, 2014. First published January 14, 2014; doi:10.1152/ajpendo.00515.2013.-Microscopic examination of transplanted islets in an ectopic environment provides information to evaluate islet engraftment, including revascularization and reinnervation. However, because of the dispersed nature of blood vessels and nerves, global visualization of the graft neurovascular network has been difficult. In this research we revealed the neurovascular network by preparing transparent mouse islet grafts under the kidney capsule with optical clearing to investigate the sympathetic reinnervation via three-dimensional confocal microscopy. Normoglycemic and streptozotocin-induced diabetic mice were used in syngeneic islet transplantation, with both groups maintaining euglycemia after transplantation. Triple staining of insulin/glucagon, blood vessels, and tyrosine hydroxylase (sympathetic marker) was used to reveal the graft microstructure, vasculature, and sympathetic innervation. Three weeks after transplantation, we observed perigraft sympathetic innervation similar to the peri-islet sympathetic innervation in the pancreas. Six weeks after transplantation, prominent intragraft, perivascular sympathetic innervation was achieved, resembling the pancreatic intraislet, perivascular sympathetic innervation in situ. Meanwhile, in diabetic recipients, a higher graft sympathetic nerve density was found compared with grafts in normoglycemic recipients, indicating the graft neural plasticity in response to the physiological difference of the recipients and the resolving power of this imaging approach. Overall, this new graft imaging method provides a useful tool to identify the islet neurovascular complex in an ectopic environment to study islet engraftment. islet transplantation; graft innervation; neural plasticity; sympathetic nerves; three dimensional ISLET TRANSPLANTATION HAS BEEN PROPOSED as an effective cure for patients with unstable type 1 diabetes in whom the diabetes care alone is inadequate to avoid serious complications (2,8,15,25). To achieve long-term graft survival, revascularization and reinnervation of islets in the new microenvironment are essential for islet engraftment (5,16,27,30,34,38). The neurovascular integration is particularly important for grafts to receive signals from the circulation and nerves in response to physiological cues to maintain glucose homeostasis and avoid hypoglycemia. However, because of the dispersed nature of blood vessels and nerves, global and integrated visualization of the graft microstructure, vasculature, and innervation has been difficult, even in animals. The difficulty of examining the neurovascular complex in a three-dimensional (3D) continuum has limited our understanding of islet engraftment after transplantation to help evaluate and improve the proce...

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Vessel painting of the microcirculation using fluorescent lipophilic tracers

Cited by 42 publications

References 10 publications

Plasticity of Schwann cells and pericytes in response to islet injury in mice

Plasticity of Schwann cells and pericytes in response to islet injury in mice

3-D imaging and illustration of the perfusive mouse islet sympathetic innervation and its remodelling in injury

Three-dimensional islet graft histology: panoramic imaging of neural plasticity in sympathetic reinnervation of transplanted islets under the kidney capsule

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