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Research article - Peer-reviewed, 2014

Laser Microdissection of Pancreatic Islets Allows for Quantitative Real-Time PCR Detection of Islet-Specific Gene Expression in Healthy and Diabetic Cats

Öhlund, Malin; Franzen, Petra; Andersson, Göran; Ström Holst, Bodil; Lau, Joey

Abstract

Background: Feline diabetes mellitus shares many similarities with human type 2 diabetes mellitus, including clinical, physiological and pathological features of the disease. The domestic cat spontaneously develops diabetes associated with insulin resistance in their middle age or later, with residual but declining insulin secretion. Humans and cats share largely the same environment and risk factors for diabetes, such as obesity and physical inactivity. Moreover, amyloid formation and loss of beta cells are found in the islets of the diabetic cat, as in humans. Altogether, the diabetic cat is a good model for type 2 diabetes in humans. The aims of the present study were to isolate feline islets using laser microdissection and to develop a quantitative method for detection of mRNA levels in islets of healthy and diabetic cats.

Results: By using the laser microdissection technique, we were able to meticulously sample islets from both healthy and diabetic cats. Insulin staining of separate sections showed many beta cells in islets from healthy cats, whereas few insulin positive cells were found in islets from diabetic cats. By quantitative real-time PCR, mRNA levels of the islet-specific genes INS, PDX1, IAPP, CHGA and IA-2 could be detected in both healthy and diabetic cats.

Conclusions: Laser microdissection allows distinct studies of islets without contamination of acinar cells. Previous attempts in isolating feline islets with different collagenase-based protocols have led to damaged islets or islets coated with exocrine acinar cells, which either way compromise the results obtained from gene expression studies. The use of the laser microdissection technique eliminates these problems as shown in this study. Differences in gene expression between healthy and diabetic cats can reveal underlying mechanisms for beta cell dysfunction and decreased beta cell mass in human type 2 diabetes.

Keywords: Beta cell; Type 2 Diabetes mellitus; Felis catus; Gene expression; Islet isolation; Laser microdissection; Pancreatic islets

Keywords

Beta cell; Type 2 Diabetes mellitus; Felis catus; Gene expression; Islet isolation; Laser microdissection; Pancreatic islets

Published in

Journal of gastroenterology, pancreatology & liver disorders
2014, volume: 1, number: 4, pages: 1-9
Publisher: Symbiosis Open Access

Authors' information

Öhlund, Malin
Swedish University of Agricultural Sciences, Department of Clinical Sciences
Franzen, Petra
Uppsala University
Swedish University of Agricultural Sciences, Department of Animal Breeding and Genetics
Swedish University of Agricultural Sciences, Department of Clinical Sciences
Lau, Joey
Uppsala University

UKÄ Subject classification

Medical Bioscience

Publication Identifiers

DOI: https://doi.org/10.15226/2374-815X/1/4/00121

URI (permanent link to this page)

https://res.slu.se/id/publ/64035