Effects of a carbohydrate restricted diet on the metabolic state and progressive pancreatic beta-cell loss in transgenic mice expressing a dominant negative GIP receptor [Elektronische Ressource] / by Martina Christine Höfer

Martina Hoefer

Sujets

• Gesundheit

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2007
• Nombre de lectures: 8
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

From the Institute of Veterinary Pathology
Department of General Pathology and Pathological Anatomy
Chair: Prof. Dr. W. Hermanns
Ludwig-Maximilians-University Munich

Under the supervision of Prof. Dr. R. Wanke




Effects of a carbohydrate restricted diet on the metabolic
state and progressive pancreatic beta-cell loss in
transgenic mice expressing a dominant negative GIP
receptor




Inaugural - Dissertation
to achieve the doctor title of veterinary medicine
at the Faculty of Veterinary Medicine of the
Ludwig-Maximilians-University, Munich


by Martina Christine Höfer
from Munich

Munich 2006 Gedruckt mit Genehmigung der Tierärztlichen Fakultät der
Ludwig-Maximilians-Universität München











Dekan: Univ.-Prof. Dr. E. P. Märtlbauer
Referent: Prof. Dr. Wanke
Korreferent: Prof. Dr. Gabius



Tag der Promotion: 09. Februar 2007














Für meine Eltern Table of content - I -
1. Introduction........................................................................................................... 1
2. Literature review................................................................................................... 3
2.1 Diabetes mellitus ............................................................................................ 3
2.1.1 Definition and description of diabetes mellitus............................................ 3
2.1.2 Classification of human diabetes mellitus................................................... 3
2.1.3 Pathophysiology of type 1 diabetes mellitus............................................... 5
2.1.4 Pathophysiology of type 2 diabetes 5
2.2 Nutrition and diabetes mellitus...................................................................... 6
2.2.1 The glycemic index (GI).............................................................................. 7
2.2.2 Dietary fiber ................................................................................................ 8
2.3 Enteroinsular axis and incretin hormones ................................................. 10
2.3.1 Definition of the enteroinsular axis............................................................ 10
2.3.2 Incretin hormones..................................................................................... 10
2.3.2.1 GIP..................................................................................................... 10
2.3.2.2 GLP-1 12
2.3.3 Incretin hormone receptors....................................................................... 13
2.3.3.1 Structure and expression of incretin hormone receptors.................... 13
2.3.3.2 Signal transduction ............................................................................ 14
2.3.3.3 Homologous desensitization of the GIP receptor............................... 14
2.3.4 The role of incretins in diabetes mellitus................................................... 15
2.3.5 Animal models for incretin research ......................................................... 17
dn2.3.5.1 GIPR transgenic mice ..................................................................... 17
-/-2.3.5.2 GIP receptor knockout (GIPR ) mice ............................................... 19
-/-2.3.5.3 GLP-1 receptor knockout (GLP-1R ) mice........................................ 21
2.3.5.4 Double Incretin Receptor Knockout Mice........................................... 22
2.3.5.5 Mt-Exendin-4 transgenic mice............................................................ 23
-/-2.3.5.6 CD26 mice and F344/DuCrj rats...................................................... 24
2.4 Apoptosis ...................................................................................................... 25
2.4.1 Definition of apoptosis .............................................................................. 25
2.4.2 Apoptosis compared to necrosis............................................................... 26
2.4.3 Mechanisms of apoptosis ......................................................................... 26
2.4.3.1 Death signals ..................................................................................... 26
2.4.3.2 Caspases........................................................................................... 26 Table of content - II -

2.4.3.3 Death receptor pathway..................................................................... 27
2.4.3.4 Mitochondrial pathway ....................................................................... 28
2.4.4 Regulatory mechanisms ........................................................................... 29
2.4.4.1 Bcl-2 family ........................................................................................ 29
2.4.4.2 IAPs and Smac/Diablo 30
2.4.5 Role of apoptosis in the endocrine pancreas............................................ 30
2.4.5.1 Physiological role of apoptosis........................................................... 30
2.4.5.2 Role of apoptosis in type 1 diabetes mellitus ..................................... 31
2.4.5.3 Role of apoptosis in type 2 diabetes mellitus 32
2.4.6 Beta-cell death in animal models of type 2 diabetes mellitus ................... 34
2.5 Renewal of the beta-cell mass ..................................................................... 35
2.5.1 Introduction............................................................................................... 35
2.5.2 Animal models for the study of beta-cell renewal ..................................... 36
2.5.2.1 IGF-II transgenic mice........................................................................ 36
2.5.2.2 GLP-1 and Exendin-4 infusion of diabetic rodents............................. 36
2.5.2.3 Glucose administration ...................................................................... 36
2.5.2.4 Pancreatectomy................................................................................. 37
2.5.3 Locations of putative stem cells/progenitor cells ...................................... 37
2.5.4 Other mechanisms of islet-cell regeneration ............................................ 39
3. Animals, materials and methods....................................................................... 41
3.1 Transgenic mice............................................................................................ 41
3.2 Materials ........................................................................................................ 43
3.2.1 Antibodies................................................................................................. 43
3.2.2 Chemicals 43
3.2.3 Enzymes and other reagents.................................................................... 44
3.2.4 Kits ........................................................................................................... 45
3.2.5 Molecular weight standards for DNA and RNA......................................... 45
3.2.6 Equipment ................................................................................................ 45
3.2.7 Composition of buffers.............................................................................. 46
3.2.7.1 Buffer for molecular biological methods ............................................. 46
3.2.7.1.1 Cutting buffer............................................................................... 46
3.2.7.1.2 DEPC-H O (0.1 %)...................................................................... 47 2
3.2.7.1.3 10x Dnase I-buffer....................................................................... 47 Table of content - III -

3.2.7.1.4 Proteinase K solution .................................................................. 47
3.2.7.1.5 TE-buffer ..................................................................................... 47
3.2.7.2 Buffer for Agarose gel electrophoresis............................................... 47
3.2.7.2.1 50x TAE-buffer ............................................................................ 47
3.2.7.2.2 1x ............................................................................... 47
3.2.7.2.3 10x TBE-buffer 47
3.2.7.2.4 1x .............................................................................. 48
3.2.7.3 Buffers for embedding and immunohistological procedures .............. 48
3.2.7.3.1 Solution A.................................................................................... 48
3.2.7.3.2 TBS (Tris-Buffer-Saline) stock solution (ph 7.6/0.05M) ............... 48
3.2.7.3.3 TBS ............................................................................................. 48
3.3 Identification of transgenic mice by PCR ................................................... 48
3.3.1 Primers..................................................................................................... 48
3.3.2 DNA isolation............................................................................................ 49
3.3.3 PCR.......................................................................................................... 49
3.3.4 Gel electrophoresis............................