Change search
ReferencesLink to record
Permanent link

Direct link
Optical projection tomography based 3D-spatial and quantitative assessments of the diabetic pancreas
Umeå University, Faculty of Medicine, Umeå Centre for Molecular Medicine (UCMM).
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The gastrointestinal tract comprises a number of digestive organs including the stomach and pancreas. The stomach is involved in the digestion and short term storage of food while the pancreas is a mixed endocrine and exocrine gland which provides the body with hormones and enzymes essential for nutritional utilisation. The pancreas consists of three different cell lineages, acinar, ductal and endocrine cells. The endocrine cells, organised in the islets of Langerhans, are scattered throughout the exocrine parenchyma and regulate blood glucose levels by production of hormones such as glucagon and insulin.

The Nkx family of homeodomain proteins controls numerous processes during development. Previous studies have identified two members belonging to the Nkx6 subfamily of Nkx proteins, Nkx6.1 and Nkx6.2. We have described the cloning and embryonic expression pattern of Nkx6.3. All three members of the Nkx6 gene family were shown to be expressed in partially overlapping domains during the development of the gastrointestinal tract and the central nervous system. Nkx6.2 was also identified as a transient marker for pancreatic exocrine cells.

Analysing gene expression patterns and morphological features in tissues and organs is often performed by stereologic sampling which is a labour-intensive two dimensional approach that rely on certain assumptions when calculating e.g. β-cell mass and islet number in the pancreas. By combined improvements in immunohistochemical protocols, computational processing and tomographic scanning, we have developed a methodology based on optical projection tomography (OPT) allowing for 3D visualisation and quantification of specifically labelled objects within intact adult mouse organs. In the pancreas, this technique allows for spatial and quantitative measurements of total islet number and β-cell mass. We have further developed a protocol allowing for high resolution regional analyses based on global OPT assessments of the pancreatic constitution. This methodology is likely to facilitate detailed cellular and molecular analysis of user defined regions of interest in the pancreas, at the same time providing information on the overall disease state of the gland.

Type 1 diabetes mellitus (T1D) can occur at any age and is characterized by the marked inability of the pancreas to secrete insulin due to an autoimmune destruction of the insulin producing β-cells. Information on the key cellular and molecular events underlying the recruitment of lymphocytes, their infiltration of the islets of Langerhans and consequent β-cell destruction is essential for understanding the pathogenesis of T1D. Using the developed methodology we have recorded the spatial and quantitative distribution of islet β-cells and infiltrating lymphocytes in the non obese diabetic (NOD) mouse model for T1D. This study shows that the smaller islets, which are predominantly organised in the periphery of the organ, are the first to disappear during the progression of T1D. The larger islets appear more resistant and our data suggest that a compensatory proliferative process is going on side by side with the autoimmune-induced β-cell destruction. Further, the formation of structures resembling tertiary lymphoid organs (TLOs) in areas apparently unaffected by insulitis suggests that local factors may provide cues for the homing of these lymphocytes back to the pancreas.

Place, publisher, year, edition, pages
Umeå: Umeå centrum för molekylär medicin (UCMM) , 2008. , 53 p.
Umeå University medical dissertations, ISSN 0346-6612 ; 1232
Keyword [en]
Nkx, development, optical projection tomography, 3D visualisation, pancreas, beta-cell mass, insulitis, diabetes
National Category
Biochemistry and Molecular Biology
URN: urn:nbn:se:umu:diva-1939ISBN: 978-91-7264-700-8OAI: diva2:142505
Public defence
2008-12-13, Betula, 6M, Norrlands Universitetssjukhus, Umeå, 09:00 (English)
Available from: 2008-11-25 Created: 2008-11-25 Last updated: 2011-04-05Bibliographically approved
List of papers
1. Cloning and analysis of Nkx6.3 during CNS and gastrointestinal development
Open this publication in new window or tab >>Cloning and analysis of Nkx6.3 during CNS and gastrointestinal development
Show others...
2006 In: Gene expression patterns, ISSN 1567-133X, Vol. 6, no 2, 162-170 p.Article in journal (Refereed) Published
urn:nbn:se:umu:diva-3688 (URN)
Available from: 2008-11-25 Created: 2008-11-25Bibliographically approved
2. Tomographic molecular imaging and 3D quantification within adult mouse organs.
Open this publication in new window or tab >>Tomographic molecular imaging and 3D quantification within adult mouse organs.
Show others...
2007 (English)In: Nature Methods, ISSN 1548-7091, Vol. 4, no 1, 31-33 p.Article in journal (Refereed) Published
Animals, Diabetes Mellitus; Type 1/*diagnosis/physiopathology, Disease Models; Animal, Disease Progression, Image Processing; Computer-Assisted/*methods, Imaging; Three-Dimensional/*methods, Islets of Langerhans/*pathology, Mice, Pancreas/*pathology, Sensitivity and Specificity, Tomography/*methods
urn:nbn:se:umu:diva-5821 (URN)10.1038/nmeth985 (DOI)17143281 (PubMedID)
Available from: 2007-11-30 Created: 2007-11-30 Last updated: 2009-05-14Bibliographically approved
3. High-resolution three-dimensional imaging of islet-infiltrate interactions based on optical projection tomography assessments of the intact adult mouse pancreas
Open this publication in new window or tab >>High-resolution three-dimensional imaging of islet-infiltrate interactions based on optical projection tomography assessments of the intact adult mouse pancreas
Show others...
2008 (English)In: Journal of Biomedical Optics, ISSN 1083-3668, Vol. 13, no 5, 054070- p.Article in journal (Refereed) Published
Abstract [en]

A predicament when assessing the mechanisms underlying the pathogenesis of type-1 diabetes (T1D) has been to maintain simultaneous global and regional information on the loss of insulin-cell mass and the progression of insulitis. We present a procedure for high-resolution 3-D analyses of regions of interest (ROIs), defined on the basis of global assessments of the 3-D distribution, size, and shape of molecularly labeled structures within the full volume of the intact mouse pancreas. We apply a refined protocol for optical projection tomography (OPT)-aided whole pancreas imaging in combination with confocal laser scanning microscopy of site-directed pancreatic microbiopsies. As such, the methodology provides a useful tool for detailed cellular and molecular assessments of the autoimmune insulitis in T1D. It is anticipated that the same approach could be applied to other areas of research where 3-D molecular distributions of both global and regional character is required.

Place, publisher, year, edition, pages
Bellingham, WA: SPIE--the International Society for Optical Engineering, 2008
biological organs, biomedical optical imaging, cellular biophysics, diseases, image resolution, laser applications in medicine, molecular biophysics, optical tomography, optical projection tomography, confocal microscopy, pancreas, microbiopsy, insulitis, diabetes
urn:nbn:se:umu:diva-23351 (URN)10.1117/1.3000430 (DOI)19021448 (PubMedID)
Available from: 2009-06-12 Created: 2009-06-12 Last updated: 2011-04-05Bibliographically approved
4. Quantification and 3-D imaging of the insulitis-induced destruction of β-cells in murine type 1 diabetes
Open this publication in new window or tab >>Quantification and 3-D imaging of the insulitis-induced destruction of β-cells in murine type 1 diabetes
Show others...
2010 (English)In: Diabetes, ISSN 0012-1797, E-ISSN 1939-327X, Vol. 59, no 7, 1756-1764 p.Article in journal (Refereed) Published
Abstract [en]

Objective: The aim of this study was to refine the information regarding the quantitative and spatial dynamics of infiltrating lymphocytes and remaining beta-cell volume during the progression of type 1 diabetes in the NOD mouse model of the disease.

Research design and methods: Using an ex vivo technique, optical projection tomography (OPT), we quantified and assessed the 3D spatial development and progression of insulitis and beta-cell destruction in pancreas from diabetes prone NOD and non-diabetes prone congenic NOD.H-2b mice between 3 and 16 weeks of age.

Results: Together with results showing the spatial dynamics of the insulitis process we provide data of beta-cell volume distributions down to the level of the individual islets and throughout the pancreas during the development and progression of type 1 diabetes. Our data provide evidence for a compensatory growth potential of the larger insulin(+) islets during the later stages of the disease around the time point for development of clinical diabetes. This is in contrast to smaller islets, which appear less resistant to the autoimmune attack. We also provide new information on the spatial dynamics of the insulitis process itself, including its apparently random distribution at onset, the local variations during its further development, and the formation of structures resembling tertiary lymphoid organs at later phases of insulitis progression.

Conclusions: Our data provides a powerful tool for phenotypic analysis of genetic and environmental effects on type 1 diabetes etiology as well as for evaluating the potential effect of therapeutic regimes.

pancreas, mouse, mice
National Category
Endocrinology and Diabetes
urn:nbn:se:umu:diva-33656 (URN)10.2337/db09-1400 (DOI)000279615100025 ()20393145 (PubMedID)
Available from: 2010-04-30 Created: 2010-04-30 Last updated: 2014-10-07Bibliographically approved

Open Access in DiVA

fulltext(1368 kB)2044 downloads
File information
File name FULLTEXT01.pdfFile size 1368 kBChecksum MD5
Type fulltextMimetype application/pdf

Search in DiVA

By author/editor
Alanentalo, Tomas
By organisation
Umeå Centre for Molecular Medicine (UCMM)
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 2044 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 685 hits
ReferencesLink to record
Permanent link

Direct link