Skip to main content
  • More from ADA
    • Diabetes Care
    • Clinical Diabetes
    • Diabetes Spectrum
    • ADA Standards of Medical Care in Diabetes
    • ADA Scientific Sessions Abstracts
    • BMJ Open Diabetes Research & Care
  • Subscribe
  • Log in
  • Log out
  • My Cart
  • Follow ada on Twitter
  • RSS
  • Visit ada on Facebook
Diabetes

Advanced Search

Main menu

  • Home
  • Current
    • Current Issue
    • Online Ahead of Print
    • ADA Scientific Sessions Abstracts
  • Browse
    • By Topic
    • Issue Archive
    • Saved Searches
    • ADA Scientific Sessions Abstracts
    • Diabetes COVID-19 Article Collection
    • Diabetes Symposium 2020
  • Info
    • About the Journal
    • About the Editors
    • ADA Journal Policies
    • Instructions for Authors
    • Guidance for Reviewers
  • Reprints/Reuse
  • Advertising
  • Subscriptions
    • Individual Subscriptions
    • Institutional Subscriptions and Site Licenses
    • Access Institutional Usage Reports
    • Purchase Single Issues
  • Alerts
    • E­mail Alerts
    • RSS Feeds
  • Podcasts
    • Diabetes Core Update
    • Special Podcast Series: Therapeutic Inertia
    • Special Podcast Series: Influenza Podcasts
    • Special Podcast Series: SGLT2 Inhibitors
    • Special Podcast Series: COVID-19
  • Submit
    • Submit a Manuscript
    • Submit Cover Art
    • ADA Journal Policies
    • Instructions for Authors
    • ADA Peer Review
  • More from ADA
    • Diabetes Care
    • Clinical Diabetes
    • Diabetes Spectrum
    • ADA Standards of Medical Care in Diabetes
    • ADA Scientific Sessions Abstracts
    • BMJ Open Diabetes Research & Care

User menu

  • Subscribe
  • Log in
  • Log out
  • My Cart

Search

  • Advanced search
Diabetes
  • Home
  • Current
    • Current Issue
    • Online Ahead of Print
    • ADA Scientific Sessions Abstracts
  • Browse
    • By Topic
    • Issue Archive
    • Saved Searches
    • ADA Scientific Sessions Abstracts
    • Diabetes COVID-19 Article Collection
    • Diabetes Symposium 2020
  • Info
    • About the Journal
    • About the Editors
    • ADA Journal Policies
    • Instructions for Authors
    • Guidance for Reviewers
  • Reprints/Reuse
  • Advertising
  • Subscriptions
    • Individual Subscriptions
    • Institutional Subscriptions and Site Licenses
    • Access Institutional Usage Reports
    • Purchase Single Issues
  • Alerts
    • E­mail Alerts
    • RSS Feeds
  • Podcasts
    • Diabetes Core Update
    • Special Podcast Series: Therapeutic Inertia
    • Special Podcast Series: Influenza Podcasts
    • Special Podcast Series: SGLT2 Inhibitors
    • Special Podcast Series: COVID-19
  • Submit
    • Submit a Manuscript
    • Submit Cover Art
    • ADA Journal Policies
    • Instructions for Authors
    • ADA Peer Review
Islet Studies

Pancreatic Islet Production of Vascular Endothelial Growth Factor-A Is Essential for Islet Vascularization, Revascularization, and Function

  1. Marcela Brissova1,
  2. Alena Shostak1,
  3. Masakazu Shiota2,
  4. Peter O. Wiebe2,
  5. Greg Poffenberger1,
  6. Jeannelle Kantz2,
  7. Zhongyi Chen1,
  8. Chad Carr1,
  9. W. Gray Jerome34,
  10. Jin Chen45,
  11. H. Scott Baldwin6,
  12. Wendell Nicholson1,
  13. David M. Bader7,
  14. Thomas Jetton8,
  15. Maureen Gannon12 and
  16. Alvin C. Powers129
  1. 1Division of Diabetes, Endocrinology, and Metabolism, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee
  2. 2Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, Tennessee
  3. 3Department of Pathology, Vanderbilt University Medical Center, Nashville, Tennessee
  4. 4Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, Tennessee
  5. 5Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, Tennessee
  6. 6Division of Pediatric Cardiology, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee
  7. 7Stahlman Laboratory, Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee
  8. 8Division of Endocrinology and Metabolism, Department of Medicine, University of Vermont College of Medicine, Burlington, Vermont
  9. 9Veterans Affairs Tennessee Valley Healthcare System, Nashville, Tennessee
  1. Address correspondence and reprint requests to Alvin C. Powers, Division of Diabetes, Endocrinology, and Metabolism, 715 PRB, Vanderbilt University, Nashville, TN 37232. E-mail: al.powers{at}vanderbilt.edu
Diabetes 2006 Nov; 55(11): 2974-2985. https://doi.org/10.2337/db06-0690
PreviousNext
  • Article
  • Figures & Tables
  • Suppl Material
  • Info & Metrics
  • PDF
Loading

Article Figures & Tables

Figures

  • FIG. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG. 1.

    Expression of angiogenic factors and their receptors in adult mouse pancreas. A: Colocalization of VEGF-A (red) and insulin (Ins; green) in islet β-cells. B: Colocalization of VEGF-A (red) and glucagon (Glu; green) in islet α-cells. C–F: Colocalization of VEGFR2 (R2; red) and PECAM-1 (green) in pancreatic vasculature. VEGFR2 is detected in microvessels of pancreatic islets and exocrine tissue. Phase contrast image in C corresponds to images in D–F. D: Islet is marked by insulin staining (Ins; blue). Arrows points to duct (C) and periductal capillaries (E and F). Arrows point to larger vessels (C, E, and F) where VEGFR2 expression is downregulated. Adjacent sections were stained for PECAM-1 (green) (H) and VEGFR2 (R2; red) (J). The corresponding phase contrast images are shown in G and I, respectively. Expression of VEGFR2 (R2; red) is downregulated in both larger arterial (a; arrow points to arterial vessel) and venous (v) vessels (J), which are positive for PECAM-1 (H), and remains present in periductal capillary plexus (d, duct). Arrows point to periductal capillaries (H and J). K: Colocalization of Ang-1 (red) and insulin (Ins; green) in islet β-cells. L: Colocalization of Ang-1 (red) and glucagon (Glu; green) indicates that Ang-1 is differentially expressed in islet cells. M and N: Colocalization of Tie2 (red) and VEGFR2 (R2; green) reveals differential expression of RTK receptors in pancreatic endothelium. Somatostatin (Som; blue5) labeling in islet δ-cells outlines the islet (M and N). Arrows point to a larger vessel where VEGFR2 expression is downregulated. O and P: Insulin (Ins; blue) is colocalized with PECAM-1 (green) (O). The section adjacent to that was stained with X-gal to visualize Tie1 (P). Expression of Tie1 receptor is detected in islet microvessels (islet boundaries marked by dotted line) and exocrine tissue (arrow) (P). Q and R: Expression of Tie1 is downregulated in veins (v), while it remains present in arties (a) and periductal capillary plexus (d, duct; arrow) as detected by X-gal staining (R). Corresponding phase contrast image (Q). S–X: Expression of ephrin-A1 and EphB4 in pancreas. S–U: Colocalization of Ephrin-A1 (red) and insulin (Ins; green) in islet β-cells. V–X: 3D reconstructed optical sections through pancreas (30-μm histological section) demonstrate expression of Eph4 receptor in pancreatic venules and veins (v). Arrows point to venules exiting the islet outlined by glucagon (Glu; blue) staining. Bar represents 50 μm (A–X).

  • FIG. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG. 2.

    Development of islet vasculature and establishment of islet blood flow occur concomitantly with islet formation. A–C: Colocalization of VEGF-A (red) and insulin (Ins; green). D: Colocalization of VEGFR2 (R2; red) and insulin (Ins; green). E–G: Colocalization of Ang-1 (red) and insulin (Ins; green). H: Colocalization of Tie2 (red) and glucagon (Glu; green). I–N: Blood flow in developing pancreas. Embryo at e16.5 was infused with 5 μl endothelium-binding tomato lectin (TL). I: Photograph of dissected digestive organs at e16.5. Black solid line shows boundaries of the pancreas (D, duodenum; P, pancreas; S, spleen; St, stomach). J: Micrograph of 10-μm DAPI-counterstained (blue) section from the pancreatic area marked by blue dotted rectangle (I). Lectin (TL; green) is detected in large blood vessels with visible presence of erythrocytes (I) and also in microvascular structures marked by two white dotted rectangles (arrows point to the panels showing enlargement of these two areas). Lectin+ microvascular structures are mostly found to be associated with endocrine cells. K–M: Pancreatic section with lectin-labeled vasculature (TL; green) (K) was costained subsequently with antibodies to VEGFR2 (R2; red) (L) and cocktail of antibodies to three islet hormones (insulin, glucagon, somatostatin, Endo; blue) (M). M: Overlay of lectin label with staining for VEGFR2 (L) and endocrine cells (Endo; blue). Coalescing endocrine cells (blue) are adjacent to the vessels with blood flow (vascular structures double positive for lectin [green] and VEGFR2 [red]). N: Colocalization of insulin (Ins; green), glucagon (Glu; red), and somatostatin (Som; blue) reveals distribution of individual endocrine cells in the developing islets (the same endocrine clusters as in M). O: Model of pancreatic islet vascularization. Bar in A–N represents 50 μm.

  • FIG. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG. 3.

    Reduced VEGF-A production by β-cells results in abnormal islet vasculature. A: VEGF-A production by islets from VEGFfl/fl (white), Rip-Cre;VEGFfl/wt (gray), and Rip-Cre;VEGFfl/fl (black) mice. *P < 0.05, **P < 0.001 compared with VEGFfl/fl mice; †P < 0.05 compared with Rip-Cre;VEGFfl/wt mice. B: Colocalization for insulin (green), glucagon (red), and somatostatin (blue) reveals normal islet morphology in mice with β-cell–reduced VEGF-A expression. C: Phase contrast images of islets in the pancreas (top panels) and corresponding micrographs of pancreatic vasculature labeled with fluorescein isothiocyanate–conjugated tomato lectin (bottom panels; dotted yellow line marks islet boundaries). D: 3D reconstructed optical sections through VEGFfl/fl (top panel) and Rip-Cre;VEGFfl/fl (bottom panel) pancreas (30-μm thick histological sections) demonstrate reduced number of vessels and reduced vessel size in VEGF-A–deficient islet (bottom panel). PECAM-1 (red) is colocalized with somatostatin (blue); dotted white line in both panels marks islet boundaries. Vessel density in Rip-Cre;VEGFfl/fl islet is higher around perimeter where VEGF-A production is maintained by non–β-cells. E: Vessel density in islets declines progressively with VEGF-A reduction. VEGFfl/fl (white), Rip-Cre;VEGFfl/wt (gray), and Rip-Cre;VEGFfl/fl (black) mice. **P < 0.001 compared with VEGFfl/fl mice; †P < 0.001 compared with Rip-Cre;VEGFfl/wt mice. Vessel density in exocrine tissue was normal across all three genotypes; 597 ± 22, 576 ± 19, and 584 ± 24 count/mm2 in VEGFfl/fl, Rip-Cre;VEGFfl/wt, and Rip-Cre;VEGFfl/fl mice, respectively. F: Area per vessel indicates similar reduction of vessel size and/or branching in Rip-Cre;VEGFfl/wt (gray) and Rip-Cre;VEGFfl/fl (black) islets compared with VEGFfl/fl (white) mice. **P < 0.001 compared with VEGFfl/fl mice. Rip-Cre;VEGFfl/wt and Rip-Cre;VEGFfl/fl mice were not statistically different. Area per vessel in exocrine tissue was normal across all three genotypes; 50 ± 3, 47 ± 2, and 48 ± 2 μm2 in VEGFfl/fl, Rip-Cre;VEGFfl/wt, and Rip-Cre;VEGFfl/fl mice, respectively. Bar in B–D represents 50 μm.

  • FIG. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG. 4.

    Mice with reduced production of VEGF-A by β-cells exhibit impaired glucose tolerance. Fasted male (A) and female (B) VEGFfl/fl (○), Rip-Cre;VEGFfl/wt (□), and Rip-Cre;VEGFfl/fl (▪) mice at 16 weeks of age (n = 10−12 per genotype in both male and female groups of mice) received glucose by intraperitoneal injection (2 g/kg of body wt). *P < 0.05 when Rip-Cre;VEGFfl/wt and Rip-Cre;VEGFfl/fl mice compared with VEGFfl/fl mice. C: Plasma insulin levels (males and females combined) normalized for blood glucose concentration, and 15 min intraperitoneal glucose tolerance test are reduced in Rip-Cre;VEGFfl/wt (gray, n = 10) and Rip-Cre;VEGFfl/fl (black, n = 18) mice compared with VEGFfl/fl mice (white, n = 23). **P < 0.01 compared with VEGFfl/fl mice. Rip-Cre;VEGFfl/wt and Rip-Cre;VEGFfl/fl mice were not statistically different. D: Pancreatic insulin content (males and females combined) is similar in VEGFfl/fl (white, n = 6), Rip-Cre;VEGFfl/wt (gray, n = 6), and Rip-Cre;VEGFfl/fl (black, n = 6) mice.

  • FIG. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG. 5.

    Reduced VEGF-A expression in islets impairs insulin output into vascular system. A: Insulin secretory response of isolated islets in cell perifusion system. Insulin secretion of islets isolated from VEGFfl/fl (○), Rip-Cre;VEGFfl/wt (□), and Rip-Cre;VEGFfl/fl (•) mice was analyzed in response to glucose and isobutylmethylxanthine (IBMX). The integrated response to 16.7 mmol/l glucose was 80.4 ± 24.4 ng insulin in VEGFfl/fl mice vs. 80.9 ± 29.5 ng insulin in Rip-Cre;VEGFfl/wt mice and 72.9 ± 9.6 ng insulin in Rip-Cre;VEGFfl/fl mice (n = 3; P = 0.963). The integrated response to 45 μmol/l isobutylmethylxanthine in the presence of 16.7 mmol/l glucose was 418 ± 51 ng insulin in VEGFfl/fl mice vs. 443 ± 62 ng insulin in Rip-Cre;VEGFfl/wt mice and 383 ± 32 ng insulin in Rip-Cre;VEGFfl/fl mice (n = 3; P = 0.705). B: Insulin secretory response of the in situ perfused pancreas. Insulin secretion from the pancreas of Rip-Cre;VEGFfl/fl mice (•) and their wild-type littermates (VEGFfl/fl; ○) was analyzed in response to glucose, isobutylmethylxanthine (IBMX), and arginine. The integrated response to 16.7 mmol/l glucose was 120 ± 17 ng insulin in VEGFfl/fl mice vs. 75.1 ± 15.2 ng insulin in Rip-Cre;VEGFfl/fl mice (n = 5; P = 0.043). The integrated response to 45 μmol/l isobutylmethylxanthine in the presence of 16.7 mmol/l glucose was 136 ± 27 ng insulin in VEGFfl/fl mice vs. 74.0 ± 12.9 ng insulin in Rip-Cre;VEGFfl/fl mice (n = 5; P = 0.0356). The integrated response to 20 mmol/l arginine in the presence of 16.7 mmol/l glucose was 877 ± 151 ng insulin in VEGFfl/fl mice vs. 469 ± 63 ng insulin in Rip-Cre;VEGFfl/fl mice (n = 5; P = 0.015). C: Ultrastructural changes to intra-islet capillaries in islets with diminished VEGF-A levels. Transmission electron microscopy images show intra-islet endothelial cells with adjacent islet cells of wild-type (left panel), Rip-Cre;VEGFfl/wt (middle panel), and Rip-Cre;VEGFfl/fl (right panel) mice. Arrows point to fenestrations and caveolae. Images are representative, taken from either group. BM, basement membrane; C, caveolae; F, fenestration; L, capillary lumen; SG, secretory granule; 40,000× magnification. Bar in C represents 500 nm.

  • FIG. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG. 6.

    Reduced β-cell production of VEGF-A affects revascularization of transplanted islets. Two-hundred islets from wild-type VEGFfl/fl (A, B, E, and F) or Rip-Cre;VEGFfl/fl (C, D, G, and H) mice were transplanted immediately after isolation beneath the renal capsule of NOD-SCID mice. The presence of functional graft vasculature was assessed at 7 days (VEGFfl/fl n = 3; Rip-Cre;VEGFfl/fl n = 6) and 1 month (VEGFfl/fl n = 5; Rip-Cre;VEGFfl/fl n = 6) posttransplantation (post-TX) by infusion of the endothelium-binding tomato lectin fluorescein isothiocyanate (TL; green). A, C, E, and G: Overlay of lectin label (TL) with staining for hormones produced by islet β-cells (Ins, insulin; blue) and non–β-cells (non–β, glucagon, somatostatin, and PP; red). Dotted white line marks graft boundaries; kidney cortex is beneath the dotted line. I: At 1 month posttransplantation, Rip-Cre;VEGFfl/fl islet grafts (black) have reduced vessel density compared with wild-type controls (white). ***P < 0.0001 compared with VEGFfl/fl islet transplants. J: Area per vessel indicates similar vessel size and/or branching in wild-type (white) and Rip-Cre;VEGFfl/fl (black) islet transplants. Bar in A–H represents 50 μm.

PreviousNext
Back to top

In this Issue

November 2006, 55(11)
  • Table of Contents
  • Index by Author
Sign up to receive current issue alerts
View Selected Citations (0)
Print
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for your interest in spreading the word about Diabetes.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Pancreatic Islet Production of Vascular Endothelial Growth Factor-A Is Essential for Islet Vascularization, Revascularization, and Function
(Your Name) has forwarded a page to you from Diabetes
(Your Name) thought you would like to see this page from the Diabetes web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Pancreatic Islet Production of Vascular Endothelial Growth Factor-A Is Essential for Islet Vascularization, Revascularization, and Function
Marcela Brissova, Alena Shostak, Masakazu Shiota, Peter O. Wiebe, Greg Poffenberger, Jeannelle Kantz, Zhongyi Chen, Chad Carr, W. Gray Jerome, Jin Chen, H. Scott Baldwin, Wendell Nicholson, David M. Bader, Thomas Jetton, Maureen Gannon, Alvin C. Powers
Diabetes Nov 2006, 55 (11) 2974-2985; DOI: 10.2337/db06-0690

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Add to Selected Citations
Share

Pancreatic Islet Production of Vascular Endothelial Growth Factor-A Is Essential for Islet Vascularization, Revascularization, and Function
Marcela Brissova, Alena Shostak, Masakazu Shiota, Peter O. Wiebe, Greg Poffenberger, Jeannelle Kantz, Zhongyi Chen, Chad Carr, W. Gray Jerome, Jin Chen, H. Scott Baldwin, Wendell Nicholson, David M. Bader, Thomas Jetton, Maureen Gannon, Alvin C. Powers
Diabetes Nov 2006, 55 (11) 2974-2985; DOI: 10.2337/db06-0690
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • RESEARCH DESIGN AND METHODS
    • RESULTS
    • DISCUSSION
    • Acknowledgments
    • Footnotes
    • REFERENCES
  • Figures & Tables
  • Suppl Material
  • Info & Metrics
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • Glucagon Resistance and Decreased Susceptibility to Diabetes in a Model of Chronic Hyperglucagonemia
  • Acyl-Ghrelin Influences Pancreatic β-Cell Function by Interference with KATP Channels
  • Pancreatic β-Cell–Specific Deletion of VPS41 Causes Diabetes Due to Defects in Insulin Secretion
Show more Islet Studies

Similar Articles

Navigate

  • Current Issue
  • Online Ahead of Print
  • Scientific Sessions Abstracts
  • Collections
  • Archives
  • Submit
  • Subscribe
  • Email Alerts
  • RSS Feeds

More Information

  • About the Journal
  • Instructions for Authors
  • Journal Policies
  • Reprints and Permissions
  • Advertising
  • Privacy Policy: ADA Journals
  • Copyright Notice/Public Access Policy
  • Contact Us

Other ADA Resources

  • Diabetes Care
  • Clinical Diabetes
  • Diabetes Spectrum
  • Scientific Sessions Abstracts
  • Standards of Medical Care in Diabetes
  • BMJ Open - Diabetes Research & Care
  • Professional Books
  • Diabetes Forecast

 

  • DiabetesJournals.org
  • Diabetes Core Update
  • ADA's DiabetesPro
  • ADA Member Directory
  • Diabetes.org

© 2021 by the American Diabetes Association. Diabetes Print ISSN: 0012-1797, Online ISSN: 1939-327X.