Noninvasive in vivo imaging of pancreatic islet cell biology

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Abstract

Advanced imaging techniques have become a valuable tool in the study of complex biological processes at the cellular level in biomedical research. Here, we introduce a new technical platform for noninvasive in vivo fluorescence imaging of pancreatic islets using the anterior chamber of the eye as a natural body window. Islets transplanted into the mouse eye engrafted on the iris, became vascularized, retained cellular composition, responded to stimulation and reverted diabetes. Laser-scanning microscopy allowed repetitive in vivo imaging of islet vascularization, beta cell function and death at cellular resolution. Our results thus establish the basis for noninvasive in vivo investigations of complex cellular processes, like beta cell stimulus-response coupling, which can be performed longitudinally under both physiological and pathological conditions.

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Figure 1: Pancreatic islet transplantation into the anterior chamber of the eye.
Figure 2: Noninvasive imaging of RIP-GFP islet engraftment and vascularization.
Figure 3: Pancreatic islets engrafted in the anterior chamber of the eye maintain glucose homeostasis.
Figure 4: Noninvasive in vivo imaging of beta cell [Ca2+]i handling.
Figure 5: Noninvasive in vivo imaging of beta cell death.

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Acknowledgements

This study was supported by grant DK-58508 and DK-075487 (to A.C.) from the US National Institutes of Health, Juvenile Diabetes Research Foundation International grant 3-2007-73 (to S.S.) and 4-2004-361, the Swedish Research Council, the Novo Nordisk Foundation, Karolinska Institutet, the Swedish Diabetes Association, The Family Knut and Alice Wallberg Foundation, Eurodia (FP6-518153), European Foundation for the Study of Diabetes, the European Foundation for the Study of Diabetes–Lilly Research Program, Berth von Kantzow's Foundation, the Family Erling-Persson Foundation and the Diabetes Research Institute Foundation.

Author information

S.S., D.N. and A.C. developed the experimental transplantation platform. S.S., D.N., O.C., J.Y., R.D.M., A.P., T.M., M.K., B.L. and A.C. did the experiments. J.W. was responsible for generating the transgenic mice. C.R. was involved in designing the transplantation protocols and writing the manuscript. S.S., D.N., I.B.L. and P.-O.B. were responsible for designing the overall experimental plan and writing the manuscript. P.-O.B. was the originator of the idea of using the anterior chamber of the eye for noninvasive in vivo imaging of pancreatic islet cell biology.

Correspondence to Alejandro Caicedo or Per-Olof Berggren.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1 and Supplementary Methods (PDF 103 kb)

Supplementary Video 1

Visualization of beta cells and blood vessels in engrafted islets in the anterior chamber of the eye. The movie shows a representative animated 3D reconstruction of the vascularization of an engrafted RIP-GFP islet in the anterior chamber of the eye. Intravenously applied Texas Red fluorescence (red) and GFP fluorescence of beta-cells (green) are observed. (MOV 2224 kb)

Supplementary Video 2

Ca2+ dye loading of islet grafts in the anterior chamber of the eye. The movie shows a representative animated three-dimensional reconstruction of two pancreatic islets engrafted in the anterior chamber of the eye. The reflection image of the islets is displayed in gray scale. Fura-Red and Fluo-4 loading of the islets in red and green, respectively. (MOV 2083 kb)

Supplementary Video 3

In vivo recording of cytoplasmic free Ca2+ concentration changes. The movie shows a representative experiment with cells of a pancreatic islet engrafted in the anterior chamber of the eye loaded with Fluo-4 (green) and Fura-Red (red). Systemic stimulation with 1 mg/kg glibenclamide is applied at 3 min. (MOV 2010 kb)

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Speier, S., Nyqvist, D., Cabrera, O. et al. Noninvasive in vivo imaging of pancreatic islet cell biology. Nat Med 14, 574–578 (2008) doi:10.1038/nm1701

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