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Optical projection tomography based 3D-spatial and quantitative assessments of the diabetic pancreas [Elektronisk resurs]

Alanentalo, Tomas, 1978- (författare)
Ahlgren, Ulf (preses)
Scharfmann, Raphael (opponent)
Umeå universitet Medicinska fakulteten (utgivare)
Publicerad: Umeå : Umeå centrum för molekylär medicin (UCMM), 2008
Engelska 53
Serie: Umeå University medical dissertations, 0346-6612 0346-6612
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  • E-bokAvhandling(Diss. (sammanfattning) Umeå : Umeå universitet, 2008)
Sammanfattning Ämnesord
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  • 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. 

Ämnesord

Natural Sciences  (hsv)
Biological Sciences  (hsv)
Biochemistry and Molecular Biology  (hsv)
Naturvetenskap  (hsv)
Biologiska vetenskaper  (hsv)
Biokemi och molekylärbiologi  (hsv)
NATURAL SCIENCES  (svep)
Biology  (svep)
Cell and molecular biology  (svep)
NATURVETENSKAP  (svep)
Biologi  (svep)
Cell- och molekylärbiologi  (svep)

Genre

government publication  (marcgt)

Indexterm och SAB-rubrik

Nkx
development
optical projection tomography
3D visualisation
pancreas
beta-cell mass
insulitis
diabetes
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