Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:355-epub-292963 Inhaltsverzeichnis |
| Beschreibung: | 125 S. Ill., graph. Darst. |
Internformat
MARC
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| 245 | 1 | 0 | |a Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina |c von Barbara Maria Braunger |
| 264 | 1 | |c 2013 | |
| 300 | |a 125 S. |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
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| adam_text | TABLE OF CONTENTS
1 INTRODUCTION: 1
1.1 RETINA 1
1.1.1 RETINAL NEURONS AND GLIA CELLS 1
1.1.1.1 RETINAL GANGLION CELLS 1
1.1.1.2 THE INNER NUCLEAR LAYER (INL): 2
1.1.1.3 NEURONS OF THE INL: AMACRINE CELLS, BIPOLAR CELLS AND HORIZONTAL
CELLS 2
1.1.1.4 RETINAL GLIA CELLS: ASTROCYTES, MIITLER GLIA CELLS AND MICROGLIA
3
1.1.1.5 CELLS OF THE OUTER NUCLEAR LAYER (ONL): PHOTORECEPTORS 4
1.1.1.6 THE RETINAL PIGMENT EPITHELIUM 6
1.1.2 PHOTOTRANSDUCTION AND VISUAL CYCLE 7
1.1.2.1 PHOTOTRANSDUCTION 7
1.1.2.2 VISUAL CYCLE 8
1.1.3 ELECTROPHYSIOLOGY: ELECTRORETINOGRAPHY (ERG) 9
1.1.4 RETINAL VESSELS IN DEVELOPMENT AND ADULTHOOD 10
1.2 NORRIN 12
1.2.1 NORRIE DISEASE: SYMPTOMS AND GENETICS 12
1.2.2 THE PROTEIN NORRIN 12
1.2.3 THE BIOLOGICAL FUNCTIONS OF NORRIN 13
1.2.3.1 ANGIOGENIC PROPERTIES OF NORRIN 13
1.2.3.2 NEUROPROTECTIVE PROPERTIES OF NORRIN 14
1.2.3.3 NORRIN IN REPRODUCTION AND DEVELOPMENT 15
1.2.4 NORRIN SIGNALING AND THE WNT/FJ-CATENIN PATHWAY 16
1.3 ANIMAL MODELS TO INDUCE CELL SPECIFIC STRESS 19
1.3.1 THE RETINOPATHIA OF PREMATURITY (ROP) MODEL 19
1.3.2 A DAMAGE MODEL FOR PHOTORECEPTOR DEGENERATION: LIGHT DAMAGE 19
1.3.3 NEUROPROTECTIVE SIGNALING PATHWAYS IN THE CONTEXT OF PHOTORECEPTOR
DEGENERATION.. 19
1.4 AIM OF THE CURRENT WORK 22
2 RESULTS 25
2.1 CHARACTERIZATION OF RPE65-NORRIN-2 MICE 25
2.1.1 SOUTHERN BLOT: SUCCESSFUL GENOMIC INSERTION OF THE TRANSGENE
CONSTRUCT 25
2.1.2 NORTHERN BLOT ANALYSIS: EFFECTIVE TRANSCRIPTION OF THE CONSTRUCT
26
2.1.3 WESTERN BLOT ANALYSIS: EFFECTIVE PROTEIN TRANSLATION 27
2.1.4 IMMUNOHISTOCHEMISTRY: RETINAL LOCALIZATION OF THE TRANSGENIC
NORRIN PROTEIN 27
HTTP://D-NB.INFO/1047649829
2.1.5 RPE65-NORRIN-2 MICE AND WNT/P-CATENIN ACTIVATION 28
2.2 PHENOTYPE ANALYSIS OF RPE65-NORRIN-2 ANIMALS 31
2.2.1 RETINAL MORPHOLOGY 31
2.2.2 ULTRASTRUCTURE OF RPE65-NORRIN-2 ANIMALS 32
2.2.3 VASCULAR PHENOTYPE 33
2.3 THE RETINOPATHIA OF PREMATURITY (ROP) MODEL 35
2.4 LIGHT DAMAGE EXPERIMENTS 40
2.4.1 MORPHOMETRY ANALYSIS 40
2.4.2 IN VIVO SLO AND OCT
IMAGING BEFORE AND AFTER LIGHT DAMAGE 43
2.4.3 FUNCTIONAL ANALYSIS: ELECTRORETINOGRAPHY (ERG) ANALYSIS 45
2.4.4 ANALYSIS OF THE VISUAL CYCLE 47
2.4.5 LIGHT EXPOSURE AND APOPTOSIS 49
2.4.6 WNT-/P-CATENIN SIGNALING AND NORRIN MEDIATED NEUROPROTECTION 51
2.5 NEUROPROTECTIVE FACTORS 54
2.5.1 TRANSGENIC NORRIN OVEREXPRESSION AND NEUROPROTECTIVE FACTORS 54
2.5.2 CORRELATION BETWEEN NORRIN INDUCED WNT/FJ-CATENIN ACTIVATION, EDN2
UPREGULATION
AND LIGHT INDUCED APOPTOSIS 58
2.5.3 BRAIN-DERIVED NEUROTROPHIC FACTOR AND THE PROTECTIVE EFFECTS OF
NORRIN 61
3 DISCUSSION 67
3.1 NORRIN IN THE CONTEXT OF NEUROPROTECTION 67
3.1.1 CHARACTERIZATION AND MORPHOLOGY OF RPE65-NORRIN-2 MICE 67
- 3.1.2 NORRIN AND LIGHT EXPOSURE 68
3.1.3 NORRIN, WNT/P-CATENIN AND EDN2 SIGNALING 69
3.1.4 NORRIN NEUROPROTECTION AND BDNF AFTER LIGHT EXPOSURE 70
3.2 NORRIN IN THE CONTEXT OF ANGIOGENESIS 73
3.2.1 VASCULAR PHENOTYPE OF RPE65-NORRIN-2 ANIMALS 73
3.2.2 NORRIN IN THE ROP MODEL 74
3.3 FUTURE DIRECTIONS 75
4 SUMMARY - ZUSAMMENFASSUNG 77
4.1 SUMMARY 77
4.2 ZUSAMMENFASSUNG 78
5 MATERIAL AND METHODS 79
5.1 METHODS 79
5.1.1 ANIMAL MODELS 79
5.1.1.1 RPE65-NORRIN-2 MICE 79
5.1.1.2 |3BL-CRYSTALKN NORRIN
MICE 79
5.1.1.3 WNT-REPORTERMICE: TOPGAL MICE 79
5.1.2 DNA ANALYSIS 80
5.1.2.1 DNA ISOLATION 80
5.1.2.2 GENOTYPING: POLYMERASE CHAIN REACTION (PCR) 80
5.1.2.2.1 GENOTYPING FOR THE TRANSGENES: NORRIN AND LACZ 80
5.1.2.2.2 GENOTYPING FOR THE LEUCINE VARIANT OF RPE65 81
5.1.2.3 AGAROSE GEL ELECTROPHORESIS 82
5.1.2.4 SOUTHERN BLOT ANALYSIS 82
5.1.2.5 SOUTHERN AND NORTHERN BLOT PROBE 83
5.1.3 RNA ANALYSIS 83
5.1.3.1 RNA ISOLATION 83
5.1.3.2 COMPLEMENTARY DNA (CDNA) SYNTHESIS 84
5.1.3.3 QUANTITATIVE REALTIME RT-PCR 85
5.1.3.4 PRIMER FOR REAL-TIME RT-PCR 85
5.1.3.5 NORTHERN BLOT ANALYSIS 86
5.1.4 PROTEIN ANALYSIS 87
5.1.4.1 PROTEIN ISOLATION 87
5.1.4.2 WESTERN BLOT ANALYSIS - 87
5.1.4.3 ANTIBODIES AND BLOCKING SOLUTIONS FOR WESTERN BLOT ANALYSIS 88
5.1.4.4 APOPTOSIS: CELL DEATH DETECTION ELISA 88
5.1.5 HISTOLOGY 89
5.1.5.1 PHENOTYPE ANALYSIS 89
5.1.5.2 IMMUNOHISTOCHEMISTRY 89
5.1.5.3 ANTIBODIES FOR IMMUNOHISTOCHEMISTRY 90
5.1.5.4 APOPTOSIS: TUNEL ANALYSIS 91
5.1.6 ANIMAL EXPERIMENTS 92
5.1.6.1 OXYGEN-INDUCED RETINAL DAMAGE: THE RETINOPATHIA PRAEMATURORUM
(ROP) MODEL.. 92
5.1.6.2 LIGHT DAMAGE 92
5.1.6.3 LIGHT DAMAGE AND MORPHOMETRIC READ OUT: THE SPIDER DIAGRAM 93
5.1.6.4 INTRAVITREAL INJECTIONS 94
5.1.6.5 FUNDUS IMAGING AND ANGIOGRAPHY 94
5.1.6.6 RETINAL WHOLE MOUNTS 94
5.1.6.7 FUNCTIONAL ANALYSIS: ELECTRORETINOGRAPHY (ERG) 95
5.1.6.8 RHODOPSIN RECOVERY IN DARKNESS 95
5.1.7 STATISTICAL ANALYSIS 96
5.2 MATERIAL 97
5.2.1 CHEMICALS AND REAGENTS 97
5.2.1.1 ENZYMES AND TAQ POLYMERASE 98
5.2.1.2 KITS 99
5.2.2 CONSUMABLE SUPPLIES AND EQUIPMENT 99
5.2.2.1 CONSUMABLE SUPPLIES 99
5.2.2.2 EQUIPMENT 99
5.2.3 GELS, BUFFERS, DILUTIONS 101
5.2.3.1 BUFFERS AND DILUTIONS 101
5.2.3.1.1 HISTOLOGY 101
5.2.3.1.2 PROTEIN ANALYSIS 101
5.2.3.1.3 DNA ANALYSIS 103
5.2.3.1.4 RNA ANALYSIS 103
5.2.3.1.5 ANIMAL EXPERIMENTS 104
5.2.3.2 POLYACRYLAMIDE GEL ELECTROPHORESIS 104
6 REFERENCES 105
7 ABBREVIATIONS 115
8 FIGURE AND TABLE LEGEND 119
8.1 FIGURE LEGEND 119
8.2 TABLE LEGEND 121
9 DANKSAGUNG 123
10 ERKIARUNG 125
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| spelling | Braunger, Barbara Maria Verfasser aut Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina von Barbara Maria Braunger 2013 125 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Regensburg, Univ., Diss., 2013 Photorezeptor (DE-588)4045934-2 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 gnd rswk-swf Netzhaut (DE-588)4171512-3 gnd rswk-swf Neuroprotektivum (DE-588)4675103-8 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Netzhaut (DE-588)4171512-3 s Photorezeptor (DE-588)4045934-2 s Neuroprotektivum (DE-588)4675103-8 s Molekularbiologie (DE-588)4039983-7 s b DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:355-epub-292963 http://epub.uni-regensburg.de/29296/ Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:355-epub-292963 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027012054&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Braunger, Barbara Maria Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina Photorezeptor (DE-588)4045934-2 gnd Molekularbiologie (DE-588)4039983-7 gnd Netzhaut (DE-588)4171512-3 gnd Neuroprotektivum (DE-588)4675103-8 gnd |
| subject_GND | (DE-588)4045934-2 (DE-588)4039983-7 (DE-588)4171512-3 (DE-588)4675103-8 (DE-588)4113937-9 |
| title | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina |
| title_auth | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina |
| title_exact_search | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina |
| title_full | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina von Barbara Maria Braunger |
| title_fullStr | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina von Barbara Maria Braunger |
| title_full_unstemmed | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina von Barbara Maria Braunger |
| title_short | Molecular analysis of the neuroprotective role of Norrin for photoreceptors in the mammalian retina |
| title_sort | molecular analysis of the neuroprotective role of norrin for photoreceptors in the mammalian retina |
| topic | Photorezeptor (DE-588)4045934-2 gnd Molekularbiologie (DE-588)4039983-7 gnd Netzhaut (DE-588)4171512-3 gnd Neuroprotektivum (DE-588)4675103-8 gnd |
| topic_facet | Photorezeptor Molekularbiologie Netzhaut Neuroprotektivum Hochschulschrift |
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| work_keys_str_mv | AT braungerbarbaramaria molecularanalysisoftheneuroprotectiveroleofnorrinforphotoreceptorsinthemammalianretina |