Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis: = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Erlangen ; Nürnberg
[2018]
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | iv, 107 Seiten Illustrationen, Diagramme 21 cm |
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| 245 | 1 | 0 | |a Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis |b = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |c vorgelegt von Maria Lasierra Losada |
| 246 | 1 | 3 | |a Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| 246 | 1 | 1 | |a Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| 264 | 1 | |a Erlangen ; Nürnberg |c [2018] | |
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| 502 | |b Dissertation |c Friedrich-Alexander-Universität Erlangen-Nürnberg |d 2018 | ||
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| adam_text | TABLE OF CONTENTS
1
ABSTRACT...........................................................................................................................................................1
2
ZUSAMMENFASSUNG........................................................................................................................................
2
3
INTRODUCTION....................................................................................................................................................4
3.1 EPITHELIAL-MESENCHYMAL
TRANSITION...........................................................................................
4
3.2 Z E B
L...........................................................................................................................................
7
3.3 THE MULTIFUNCTIONAL
PANCREAS..................................................................................................
11
3.3.1 DIABETES
MELLITUS.............................................................................................................
13
3.4 PANCREAS
ORGANOGENESIS..........................................................................................................
17
3.5 EMT IN THE
PANCREAS.................................................................................................................19
4
OBJECTIVES.....................................................................................................................................................21
5
RESULTS...........................................................................................................................................................23
5.1 ZEBL EXPRESSION IN THE EMBRYONIC
PANCREAS...........................................................................23
5.2 ANALYSIS OF ZEBL DEPLETION IN THE PANCREAS
EPITHELIUM............................................................25
5.2.1 CONFIRMATION OF PROPER RECOMBINATION
EFFICIENCY.............................................................
25
5.2.2 ZEBLA/APANC MICE DISPLAY AN EXPANDED PANCREAS SIZE AT SIX MONTHS
OF AGE
....................
27
5.2.3 DEPLETION OF ZEBL FROM THE PANCREAS EPITHELIUM INCREASES ENDOCRINE
MARKERS
...........
28
5.2.4 INCREASE IN AREA OF ISLETS OF LANGERHANS AND DEFICIENT BLOOD
GLUCOSE REGULATION IN
ZEBL-DEPLETED
MICE.............................................................................................................................
31
5.2.5 ZEBLA/APANC MICE EXHIBIT INCREASED PROLIFERATION AND DECREASED
APOPTOSIS IN
THE
PANCREAS........................................................................................................................................
33
5.3 CHALLENGING THE ZEBLA/APANC
PANCREAS.......................................................................................34
5.3.1 ZEBL DEPLETION IN A DIABETIC MOUSE MODEL EXACERBATES LIVER
PATHOLOGIES
........................
35
5.3.2 ZEBL DEPLETION DOES NOT ALTER SUSCEPTIBILITY TO PANCREATITIS OR TO
TISSUE
REPAIR
RESPONSE....................................................................................................................................
39
5.4 ANALYSIS OF MODERATE ZEBL OVEREXPRESSION IN EPITHELIAL CELLS OF THE
PANCREAS IN NEWBORNS
AND DURING
EMBRYOGENESIS......................................................................................................................40
5.4.1 ZEBL LEVELS ARE 2-FOLD UPREGULATED IN THE PANCREAS OF INDZEBL MICE
.............................
40
5.4.2 ZEBL UPREGULATION IN THE PANCREAS RESULTS IN PANCREAS AGENESIS
WITH EARLY LETHALITY .... 43
5.4.3 ZEBL OVEREXPRESSION IN THE EMBRYONIC PANCREAS INDUCES APOPTOSIS
AND AN
UNDIFFERENTIATED
PHENOTYPE................................................................................................................
45
5.4.4 CELL COMMITMENT IN THE PANCREAS IS NOT AFFECTED BY EXPERIMENTALLY
INCREASED
ZEBL
EXPRESSION...................................................................................................................................
49
5.4.5 EMT-TFS ARE NOT DEREGULATED BY ZEBL INDUCTION IN THE PANCREAS
...................................
52
6
DISCUSSION....................................................................................................................................................
54
6.1 ENDOGENOUS ZEBL EXPRESSION IN THE PANCREAS EPITHELIUM IS CRITICAL
FOR ORGANOGENESIS AND
HOMEOSTASIS.............................................................................................................................................
54
6.1.1 PDXL-CRE RECOMBINATION
EFFICIENCY..................................................................................54
6.1.2 ZEBL-DEPENDENT IMPLICATIONS ON ORGAN DIFFERENTIATION
..................................................
55
6.1.3 EMT-TF ZEBL PLAYS A ROLE IN MAINTAINING PANCREATIC EPITHELIAL
HOMEOSTASIS
.................
56
6.1.4 CONTROL OF EPITHELIAL AND MESENCHYMAL MARKER GENES IN THE
EMBRYONIC AND ADULT
PANCREAS BY ZE B
L.................................................................................................................................
57
6.1.5 ABSENCE OF ZEBL EXPRESSION IN THE ENDOCRINE PRECURSOR CELLS
........................................
57
6.1.6 CHANGES IN ZEBL LEVELS DO NOT ALTER EXPRESSION OF OTHER EMT-TFS
.................................
58
6.1.7 ZEBL DEPLETION LEADS TO PANCREATIC HYPERPLASIA, WHILE
ZEBL OVEREXPRESSION RESULTS IN
PANCREATIC
HYPOPLASIA...............................................................................?.........................................59
6.2 ZEBL IN TISSUE REPAIR AND FIBROSIS DURING PANCREAS
INJURY........................................................60
6.3 ZEBL-DEPENDENT REGULATION OF ENDOCRINE
MARKERS..................................................................61
6.4 INCREASED STEATOSIS IN ZEBL-DEFICIENT
MICE..............................................................................63
7
MATERIALS......................................................................................................................................................
65
7.1 MOUSE
STRAINS..........................................................................................................................
65
7.1.1 STRAINS, TRANSGENES AND GENETICALLY MODIFIED
ALLELES.......................................................65
7.1.2 EQUIPMENT FOR MOUSE
NECROSCOPY...................................................................................66
7.1.3 ANALGESICS, ANESTHETICS AND
DIETS.....................................................................................67
7.2
OLIGONUCLEOTIDES......................................................................................................................67
7.2.1 GENOTYPING
PRIMERS.........................................................................................................67
7.2.2 QRT-PCR MRNA
PRIMERS...................................................................................................67
7.2.3 QRT-PCR MIRNA
PRIMERS..................................................................................................68
7.2.4
PLASMID..............................................................................................................................
68
7.3
ANTIBODIES.................................................................................................................................69
7.3.1 PRIMARY
ANTIBODIES...........................................................................................................
69
7.3.2 SECONDARY
ANTIBODIES......................................................................................................
70
7.4 INSULIN ELISA
KIT.........................................................................................................................70
7.5 BUFFERS, CHEMICALS, SOLUTIONS AND OTHER
REAGENTS....................................................................
71
7.5.1
GENOTYPING.......................................................................................................................71
7.5.2 MOUSE
EXPERIMENTS.........................................................................................................
71
7.5.3
HISTOLOGY...........................................................................................................................71
7.5.4 RNA/MIRNA ISOLATION AND
ANALYSIS..................................................................................
73
7.6
EQUIPMENT................................................................................................................................74
7.7
SOFTWARE...................................................................................................................................75
8
METHODS......................................................................................................................................................
76
8.1 ANIMAL
EXPERIMENTS.................................................................................................................
76
8.1.1 MOUSE
HUSBANDRY............................................................................................................
76
8.1.2
GENOTYPING......................................................................................................................
76
8.1.3 MOUSE NECROPSY AND DISSECTED
TISSUES............................................................................
78
8.1.4 EMBRYO
DISSECTION............................................................................................................
78
8.1.5 GLUCOSE TOLERANCE TEST
(GTT)..........................................................................................
78
8.1.6 INDUCTION OF TYPE 2 DIABETES
(T2D).................................................................................
79
8.1.7 INDUCTION OF CHRONIC PANCREATITIS
(CP).............................................................................
79
8.2 HISTOLOGICAL
TECHNIQUES............................................................................................................
79
8.2.1 TISSUE PROCESSING, PARAFFIN-EMBEDDING AND
SECTIONING...................................................79
8.2.2 CRYOTISSUE PROCESSING AND
SECTIONING..............................................................................80
8.2.3 HEMATOXYLIN & EOSIN (H&E)
STAINING................................................................................80
8.2.4 MASSON S TRICHROME STAINING
(MTS)................................................................................81
8.2.5 OIL RED 0 STAINING
(ORO)..................................................................................................81
8.2.6 IMMUNOHISTOCHEMISTRY (IHC) ON PARAFFIN
SECTIONS..........................................................81
III
8.2.7 INDIRECT IMMUNOFLUORESCENCE LABELING (IF) ON CRYOSECTIONS
81
8.3 HISTOLOGICAL QUANTIFICATION AND
SCORING....................................................................................82
8.3.1 INSULIN SCORING - ADULT
PANCREAS.......................................................................................82
8.3.2 KI67/DEAVED CASPASE-3 - ADULT PANCREAS
SCORING...........................................................82
8.3.3 KI67/CLEAVED CASPASE-3 - EMBRYONIC PANCREAS
SCORING..................................................82
8.3.4 EMT-TFS - EMBRYONIC PANCREAS
SCORING...........................................................................82
8.3.5 LIVER
STEATOSIS..................................................................................................................
82
8.4 MOLECULAR BIOLOGY
TECHNIQUES..................................................................................................
83
8.4.1 PANCREATIC RNA ISOLATION AND QUANTITATIVE
RT-PCR..........................................................83
8.4.2 MRNA CDNA SYNTHESIS AND
QRT-PCR................................................................................84
8.4.3 MIRNA CDNA SYNTHESIS AND
QRT-PCR:...............................................................................
85
8.4.4 INSULIN
ELISA.....................................................................................................................
86
8.5 STATISTICAL
ANALYSIS....................................................................................................................
86
9
REFERENCES...................................................................................................................................................88
10
APPENDIX.....................................................................................................................................................100
10.1 LIST OF
FIGURES......................................................................................................................
100
10.2 SUPPLEMENTARY
FIGURES......................................................................................................
101
10.2.1 LIST OF SUPPLEMENTARY
FIGURES.........................................................................................
101
10.3
ABBREVIATIONS.....................................................................................................................
104
11 ACKNOWLEDGMENTS -
AGRADECIMIENTOS..................................................................................................
106
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| id | DE-604.BV045201804 |
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| indexdate | 2024-07-10T08:11:25Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-030590759 |
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| physical | iv, 107 Seiten Illustrationen, Diagramme 21 cm |
| publishDate | 2018 |
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| spelling | Lasierra Losada, María Verfasser aut Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase vorgelegt von Maria Lasierra Losada Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase Erlangen ; Nürnberg [2018] iv, 107 Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Alexander-Universität Erlangen-Nürnberg 2018 Bauchspeicheldrüse (DE-588)4004722-2 gnd rswk-swf Homöostase (DE-588)4160575-5 gnd rswk-swf Transkriptionsfaktor (DE-588)4303350-7 gnd rswk-swf Organogenese (DE-588)4266352-0 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Transkriptionsfaktor (DE-588)4303350-7 s Bauchspeicheldrüse (DE-588)4004722-2 s Organogenese (DE-588)4266352-0 s Homöostase (DE-588)4160575-5 s DE-604 B:DE-101 application/pdf http://d-nb.info/1166465098/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030590759&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lasierra Losada, María Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase Bauchspeicheldrüse (DE-588)4004722-2 gnd Homöostase (DE-588)4160575-5 gnd Transkriptionsfaktor (DE-588)4303350-7 gnd Organogenese (DE-588)4266352-0 gnd |
| subject_GND | (DE-588)4004722-2 (DE-588)4160575-5 (DE-588)4303350-7 (DE-588)4266352-0 (DE-588)4113937-9 |
| title | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| title_alt | Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| title_auth | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| title_exact_search | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| title_full | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase vorgelegt von Maria Lasierra Losada |
| title_fullStr | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase vorgelegt von Maria Lasierra Losada |
| title_full_unstemmed | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase vorgelegt von Maria Lasierra Losada |
| title_short | Analysis of the role of Zeb1 during pancreas organogenesis and homeostasis |
| title_sort | analysis of the role of zeb1 during pancreas organogenesis and homeostasis analyse der bedeutung von zeb1 wahrend der pankreasorganogenese und homoostase |
| title_sub | = Analyse der Bedeutung von Zeb1 während der Pankreasorganogenese und -homöostase |
| topic | Bauchspeicheldrüse (DE-588)4004722-2 gnd Homöostase (DE-588)4160575-5 gnd Transkriptionsfaktor (DE-588)4303350-7 gnd Organogenese (DE-588)4266352-0 gnd |
| topic_facet | Bauchspeicheldrüse Homöostase Transkriptionsfaktor Organogenese Hochschulschrift |
| url | http://d-nb.info/1166465098/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030590759&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT lasierralosadamaria analysisoftheroleofzeb1duringpancreasorganogenesisandhomeostasisanalysederbedeutungvonzeb1wahrendderpankreasorganogeneseundhomoostase AT lasierralosadamaria analysederbedeutungvonzeb1wahrendderpankreasorganogeneseundhomoostase |
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