Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
München
Verl. Dr. Hut
2014
|
| Edition: | 1. Aufl. |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | 145 S. Ill., graph. Darst. |
| ISBN: | 9783843916653 |
Staff View
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| adam_text | TABLE OF CONTENTS
TABLE OF CONTENTS
PUBLICATIONS AND PRESENTATIONS 5
ZUSAMMENFASSUNG - ABSTRACT 7
TABLE OF CONTENTS 11
1 INTRODUCTION 15
1.1 IBD
- A PROTOTYPIC MULTIFACTORIAL
CHRONIC DEGENERATIVE LIFESTYLE DISEASE 15
1.2 MULTI-FUNCTIONAL ROLE OF THE EPITHELIUM IN MUCOSAL
HOMEOSTASIS 17
1.3 MOUSE MODELS FOR THE IMPACT OF EPITHELIAL INTEGRITY ON IBD 19
1.4 UNFOLDED PROTEIN RESPONSES FOR HOMEOSTASIS AND IN DISEASE 20
1.4.1 THE ER UNFOLDED PROTEIN RESPONSE 20
1.4.2 INVOLVEMENT OF ER UPR MECHANISMS IN DISEASE PATHOGENESES 23
1.4.3 THE MITOCHONDRIAL UNFOLDED PROTEIN
RESPONSE 25
1.4.4 THE ER - MITOCHONDRIAL SIGNALING NETWORK 27
1.5 HSP60 - CHAPERONE AND DANGER SIGNAL 29
2 AIM OF THE WORK 35
3 MATERIAL AND METHODS 37
3.1 SUPPLIER OF EXPERIMENTAL MATERIAL 37
3.2 ANIMAL MODELS 38
3.2.1 ETHICS STATEMENT AND ANIMAL HOUSING 38
3.2.2 PURCHASE OF ESTABLISHED MOUSE MODELS 38
3.2.3 GENERATION OF NEW MOUSE MODELS 39
3.2.4 EMBRYO PREPARATION AND GENOTYPING 40
3.2.5 INDUCTION OF POSTNATAL GENOMIC
RECOMBINATION AND MONITORING OF ANIMALS 40
3.2.6 NUTRIENT UPTAKES IN SMALL INTESTINAL EVERTED GUT RINGS 41
3.2.7 CALORIMETRY OF FECAL SAMPLES 41
3.2.8 USSING CHAMBER MEASUREMENTS 41
3.2.9 TISSUE PROCESSING, H&E-, ALCIAN BLUE-, IHC- AND IF STAINING, ISH
42
3.2.10 ISOLATION OF TOTAL PRIMARY IEC AND JEJUNAL VILLUS
TIP AND CRYPT BOTTOM EPITHELIUM 44
3.3 CELL LINES AND BACTERIA 45
3.3.1 MODE K 45
3.3.2 COMPETENT E. COLI 46
3.4 PLASMIDS 47
3.5 MOLECULAR CLONING 48
3.6 TRANSFECTION OF ADHERENT CELL LINES 54
3.7 CHROMATIN IMMUNOPRECIPITATION FOR ANALYSIS OF
DNA-PROTEIN-INTERACTION 55
3.8 QUANTITATIVE REAL TIME PCR 56
3.9 DISCONTINUOUS DENATURING POLYACRYLAMID
GEL ELECTROPHORESIS 58
3.10 WESTERN BLOT 59
3.11 ELECTRON MICROSCOPY 61
3.12 RNA MICROARRAY 61
3.13 STATISTICAL ANALYSIS 62
11
HTTP://D-NB.INFO/1054742057
TABLE OF CONTENTS
4 RESULTS 63
4.1 INTERRELATION OF ER- AND MT UPR SIGNALING 63
4.1.1 PROMOTER ANALYSIS OF ER- AND MT UPR RESPONSIVE GENES 63
4.1.2 CONSTRUCTION AND IN VITRO VALIDATION OF CHOP AND NATF6
OVEREXPRESSING VECTORS 64
4.2 GENERATION OF THE NATF6-HA AND CHOP-HA CONDITIONAL TRANSGENIC MOUSE
MODELS 67
4.3 GENERATION OF THE CONSTITUTIVE AND
CONDITIONAL HSP60 KNOCKOUT MOUSE MODEL 67
4.4 CHARACTERIZATION OF THE CONSTITUTIVE HSP60MOUSE MODEL 67
4.4.1 GENOTYPING 67
4.4.2 EMBRYOLETHALITY OF HSP60** MICE 68
4.4.3 PHENOTYPIC SCREENING OF SELECTED ORGANS OF HSP60 ~ MICE 68
4.4.4 BASIC CHARACTERIZATION OF THE INTESTINE OF HSP60* - MICE 69
4.4.5 HSP&Y- MICE IN AN ACUTE AN A CHRONIC MODEL OF INTESTINAL
INFLAMMATION 72
4.5 CHARACTERIZATION OF THE HSP60
UI
EC
MOUSE 76
4.5.1 GENOTYPING OF THE CONDITIONAL KNOCK OUT ALLELE 76
4.5.2 EMBRYOLETHALITY OF THE HSP60
UA,EC
MOUSE 76
4.6 CHARACTERIZATION OF THE TAMOXIFEN INDUCIBLE HSP60
S,ILEC
MOUSE 78
4.6.1 PHENOTYPIC, METABOLIC AND INTESTINAL CHARACTERIZATION OF THE HSP60
UJ,EC
MOUSE 78
4.6.2 FUNCTIONAL CHARACTERIZATION OF THE EPITHELIUM IN HSPEO*
1
-
MICE 82
4.6.3 CHARACTERIZATION OF THE EPITHELIAL STRESS RESPONSES IN HSP60^ 84
4.6.4 CHARACTERIZATION OF THE MITOCHONDRIAL APPEARANCE IN IEC OF HSP60
1/4 ,EC
MICE 88
4.6.5 EFFECTS OF HSP60 DEPLETION ON EPITHELIAL PROLIFERATION 90
4.7 CHARACTERIZATION OF A STEM CELL SPECIFIC HSP60 DEPLETION IN HSP6(Y
JMSC
MICE 94
5 DISCUSSION 97
5.1 HSP60
4BIEC
MICE - THE PARADOX OF EMBRYOLETHALITY 97
5.2 HSP60
S/I,EC
MICE - THE COMPLEXITY OF A FATAL PHENOTYPE 98
5.3 THE IMPACT OF HSP60 DEFICIENCY ON MITOCHONDRIA IN IEC 100
5.4 HSP60
!JI
EC
MICE - A MODEL FOR ACTIVATED MT UPR AND OXIDATIVE STRESS IN IEC 101
5.5 HSPSWEC MJCE _
A
MODEL FOR STEM CELL LOSS AND EPITHELIAL REGENERATION? 102
5.6 THE COMPENSATORY POTENTIAL OF HSP60 IN HAPLODEFICIENT HSP60* MICE
105
6 CONCLUSION AND PERSPECTIVE 109
7 APPENDIX 111
7.1 SEQUENCE OF HA-NATF6 AND NAFT6-HA 111
7.2 GENERATION OF THE NATF6-HA CONDITIONAL TRANSGENIC MOUSE 113
7.3 SEQUENCE OF HA-CHOP AND CHOP-HA 115
7.4 GENERATION OF THE CHOP-HA CONDITIONAL TRANSGENIC MOUSE 116
7.5 SEQUENCE OF HSP60 AND TRUNCATED HSP60 118
7.6 GENERATION OF THE HSP60 CONSTITUTIVE AND CONDITIONAL KNOCK OUT MOUSE
MODEL 119
12
TABLE OF CONTENTS
LIST OF FIGURES 123
LIST OF TABLES 125
LIST OF ABBREVIATIONS 127
LIST OF REFERENCES 129
ACKNOWLEDGEMENTS 141
CURRICULUM VITAE 143
ERKIARUNG 145
13
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| author_facet | Berger, Emanuel Clemens |
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| author_sort | Berger, Emanuel Clemens |
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| bvnumber | BV042031362 |
| classification_tum | OEK 161d |
| ctrlnum | (OCoLC)889875158 (DE-599)BVBBV042031362 |
| discipline | Ökotrophologie |
| edition | 1. Aufl. |
| format | Thesis Book |
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| indexdate | 2024-07-10T01:10:59Z |
| institution | BVB |
| isbn | 9783843916653 |
| language | English |
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| spelling | Berger, Emanuel Clemens Verfasser aut Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models Emanuel Clemens Berger 1. Aufl. München Verl. Dr. Hut 2014 145 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: München, Techn. Univ., Diss., 2014 Zsfassung in dt. Sprache Proteine (DE-588)4076388-2 gnd rswk-swf Darmepithel (DE-588)4220708-3 gnd rswk-swf Signaltransduktion (DE-588)4318717-1 gnd rswk-swf Hitzeschock-Proteine (DE-588)4255487-1 gnd rswk-swf Homöostase (DE-588)4160575-5 gnd rswk-swf Gentechnisch veränderter Organismus (DE-588)4353579-3 gnd rswk-swf Maus (DE-588)4169148-9 gnd rswk-swf Tiermodell (DE-588)4140660-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Darmepithel (DE-588)4220708-3 s Homöostase (DE-588)4160575-5 s Signaltransduktion (DE-588)4318717-1 s Hitzeschock-Proteine (DE-588)4255487-1 s Maus (DE-588)4169148-9 s Tiermodell (DE-588)4140660-6 s DE-604 Gentechnisch veränderter Organismus (DE-588)4353579-3 s Proteine (DE-588)4076388-2 s b DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027472800&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Berger, Emanuel Clemens Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models Proteine (DE-588)4076388-2 gnd Darmepithel (DE-588)4220708-3 gnd Signaltransduktion (DE-588)4318717-1 gnd Hitzeschock-Proteine (DE-588)4255487-1 gnd Homöostase (DE-588)4160575-5 gnd Gentechnisch veränderter Organismus (DE-588)4353579-3 gnd Maus (DE-588)4169148-9 gnd Tiermodell (DE-588)4140660-6 gnd |
| subject_GND | (DE-588)4076388-2 (DE-588)4220708-3 (DE-588)4318717-1 (DE-588)4255487-1 (DE-588)4160575-5 (DE-588)4353579-3 (DE-588)4169148-9 (DE-588)4140660-6 (DE-588)4113937-9 |
| title | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models |
| title_auth | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models |
| title_exact_search | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models |
| title_full | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models Emanuel Clemens Berger |
| title_fullStr | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models Emanuel Clemens Berger |
| title_full_unstemmed | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models Emanuel Clemens Berger |
| title_short | Impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models |
| title_sort | impact of unfolded protein responses on intestinal epithelial homeostasis in genetically modified mouse models |
| topic | Proteine (DE-588)4076388-2 gnd Darmepithel (DE-588)4220708-3 gnd Signaltransduktion (DE-588)4318717-1 gnd Hitzeschock-Proteine (DE-588)4255487-1 gnd Homöostase (DE-588)4160575-5 gnd Gentechnisch veränderter Organismus (DE-588)4353579-3 gnd Maus (DE-588)4169148-9 gnd Tiermodell (DE-588)4140660-6 gnd |
| topic_facet | Proteine Darmepithel Signaltransduktion Hitzeschock-Proteine Homöostase Gentechnisch veränderter Organismus Maus Tiermodell Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027472800&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT bergeremanuelclemens impactofunfoldedproteinresponsesonintestinalepithelialhomeostasisingeneticallymodifiedmousemodels |