Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2014
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 69 Bl. Ill., graph. Darst. 30 cm |
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| adam_text | TABLE OF CONTENTS
SUMMARY 1
ZUSAMMENFASSUNG 2
INTRODUCTION 3
THE MOTOR SYSTEM AND HEREDITARY SPASTIC PARAPLEGIAS 3
CEREBELLUM 4
PATHOGENIC MECHANISMS OF HEREDITARY SPASTIC PARAPLEGIAS 5
HEREDITARY SPASTIC PARAPLEGIA TYPE 15 (SPG15) 6
AIM OF THIS STUDY 9
MATERIAL AND METHODS 10
MATERIALS 10
CHEMICALS 10
MICROBIAL STRAINS AND LABORATORY ANIMALS 10
BACTERIAL VECTORS 10
ANTIBODIES 11
MICROBIOLOGY METHODS 12
PREPARATION OF CHEMO-COMPETENT BACTERIA 12
TRANSFORMATION OF COMPETENT BACTERIA 13
SCREENING OF LAMBDA PHAGE LIBRARY 13
MOLECULAR BIOLOGY METHODS 15
CLONING OF TARGETING VECTOR AND RESTRICITON DIGEST 15
BLUNTING OF STICKY ENDS AND DEPHOSPHORYLATION 15
AGAROSE GEL ELECTROPHORESIS 16
DNA LIGATION 16
DNA SEQUENCING 16
ISOLATION AND PURIFICATION OF NUCLEIC ACIDS 17
ISOLATION OF DNA FROM A-PHAGE 17
HTTP://D-NB.INFO/1047969289
MINI PREPARATION OF PLASMID DNA BACTERIAL CLONES 18
MIDI PREPARATION OF PLASMID DNA 18
SLOT LYSIS 18
PURIFICATION OF DNA FROM AN AGAROSE GEL 19
PURIFICATION OF DNA FOR ES CELL CULTURE 19
PHENOL-CHLOROPHORM EXTRACTION OF MOUSE GENOMIC DNA FROM TAIL BIOPSY 20
DNA EXCTRACTION FROM MOUSE TAIL BIOPSIES BY HOT-SHOT METHOD 20
PURIFICATION OF RNA FROM MOUSE TISSUES 21
POLYMERASE CHAIN REACTION (PCR) 21
GENOTYPING PCR 22
CELL BIOLOGY METHODS 22
MOUSE EMBRIONIC STEM CELL CULTURE 22
BLASTOCYST INJECTION AND GENERATION OF CHIMERIC MICE 23
SOUTHERN BLOT 24
NORTHERN BLOT 24
RADIO-LABELLING THE PROBES 25
NON-RADIOACTIVE IN SITU HYBRIDIZATION 25
HISTOLOGY 26
TISSUE SECTIONING 26
HISTOLOGICAL STAINING 27
HEMATOXILIN AND EOSIN STAINING 27
SUDAN BLACK STAINING 27
ULTRASTRUCUTRAL ANALYSIS 28
IMMUNOHISTOCHEMISTRY 28
PROTEIN BIOCHEMISTRY 29
PROTEIN PREPARATION FOR WESTERN BLOT ANALYSIS 29
IMMUNOBLOTTING 30
ZFYVE26-GFP OVEREXPRESSION AND CELL CULTURE 30
C0S7 CULTURE 31
CORTICAL NEURON CULTURE 31
TRANSFERRIN UPTAKE 31
BEHAVIORAL ANALYSIS 32
MORIS WATER MAZE 32
VIDEO BASED QUANTITATIVE GAIT ANALYSIS 33
RESULTS 34
EXPRESSION OF ZFYVE26 IN MURINE TISSUES 34
EXPRESSION OF ZFYVE26 IN THE BRAIN 34
SUBCELLULAR LOCALIZATION OF ZFYVE26 IN VITRO 36
GENERATION OF ZFYVE26-KO MICE 38
PHENOTYPE OF ZFYVE26-KO MICE 39
LEARNING AND MEMORY WERE NOT AFFECTED IN 5-MONTHS-OLD ZFYVE26-KO MICE 40
ZFYVE26-KO MICE SHOWED PROGRESSIVE MOVEMENT DISORDER 41
DISRUPTION OF ZFYVE26 CAUSES SEVERE NEURON LOSS IN CNS 42
NEURON LOSS AND ASTROCYTOSIS IN THE CORTEX AND CEREBELLUM OF ZFYVE26-KO
MICE 44
WHITE MATTER DEGENERATION IN THE SPINAL CORD OF ZFYVE26-KO MICE 45
ACCUMULATION OF LIPOFUSCIN-LIKE MATERIAL IN THE ZFYVE26-KO NEURONS 46
FINE STRUCTURE OF PURKINJE CELLS 48
AUTOFLUORESCENT DEPOSITS ARE LAMPL POSITIVE 48
INTERNALIZATION OF TRANSFERRIN IS NOT AFFECTED IN ZFYVE26-KO NEURONS 49
ZFYVE26-GFP IS ASSOCIATED WITH AUTOPHAGOSOMES IN VITRO 50
NO MAJOR CHANGE IN ENDOLYSOSOMAL OR AUTOPHAGY MARKERS 51
STATISTICS 52
DISCUSSION 53
EXPRESSION OF ZFYVE26 IN DIFFERENT TISSUES 53
CELLULAR FUNCTION OF ZFYVE26 53
ZFYVE26-KO MICE DEVELOPED COMPLICATED HSP 54
CELLULAR PATHOLOGY UPON DISRUPTION OF ZFYVE26 55
POSIBLE LINKS BETWEEN ENDOSOMAL LOCALIZATION AND LYSOSOMAL PATHOLOGY
UPON
DESRUPTION OFZFYVE26 55
AUTOPHAGY 57
CONCLUSIONS AND PERSPECTIES 59
LITERATURE 61
APPENDIX 66
CURRICULUM VITAE 66
PUBLICATIONS 67
ACKNOWLEDGEMENTS 68
EHRENWORTLICHE ERKLARUNG 69
|
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| spelling | Khundadze, Mukhran 1984- Verfasser (DE-588)1047969068 aut Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 von Mukhran Khundadze 2014 69 Bl. Ill., graph. Darst. 30 cm txt rdacontent n rdamedia nc rdacarrier Jena, Univ., Diss., 2014 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027449870&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Khundadze, Mukhran 1984- Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 |
| subject_GND | (DE-588)4113937-9 |
| title | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 |
| title_auth | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 |
| title_exact_search | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 |
| title_full | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 von Mukhran Khundadze |
| title_fullStr | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 von Mukhran Khundadze |
| title_full_unstemmed | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 von Mukhran Khundadze |
| title_short | Generation and characterization of a mouse model for hereditary spastic paraplegia SPG15 |
| title_sort | generation and characterization of a mouse model for hereditary spastic paraplegia spg15 |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027449870&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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