Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Köln
2018
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | 170, XVI Seiten Illustrationen 21 cm |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV045423307 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | t | ||
| 008 | 190124s2018 gw a||| m||| 00||| eng d | ||
| 015 | |a 18,H08 |2 dnb | ||
| 016 | 7 | |a 1161869433 |2 DE-101 | |
| 020 | |c Broschur | ||
| 035 | |a (OCoLC)1098204779 | ||
| 035 | |a (DE-599)DNB1161869433 | ||
| 040 | |a DE-604 |b ger |e rda | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c XA-DE | ||
| 049 | |a DE-355 |a DE-83 | ||
| 084 | |a 570 |2 sdnb | ||
| 100 | 1 | |a Schneider, Svenja |d 1986- |e Verfasser |0 (DE-588)116006492X |4 aut | |
| 245 | 1 | 0 | |a Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models |c vorgelegt von Svenja Schneider aus Oldenburg |
| 264 | 1 | |a Köln |c 2018 | |
| 300 | |a 170, XVI Seiten |b Illustrationen |c 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Universität zu Köln |d 2018 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u http://d-nb.info/1161869433/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030809154&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-030809154 | ||
Datensatz im Suchindex
| _version_ | 1804179304654831616 |
|---|---|
| adam_text | TABLE OF CONTENT
TABLE OF
CONTENT.......................................................................................................................
I
LIST OF
FIGURES.............................................................................................................................V
LIST OF
TABLES..............................................................................................................................VI
ABBREVIATIONS..................................................................................................................
VII
S U M M A R
Y...................................................................................................................................VIII
ZUSAMMENFASSUNG...................................................................................................................
X
1.
INTRODUCTION..................................................................................................................
1
1.1 PROXIMAL SPINAL MUSCULAR ATROPHY - PATHOGENESIS, CLASSIFICATION AND
GENETICS
........
1
1.1.1 SMA
PATHOGENESIS.........................................................................................................
2
1.1.2 SMA
CLASSIFICATION..........................................................................................................
3
1.1.3 MOLECULAR GENETICS OF S M A
.........................................................................................
5
1.2 THE SMN
PROTEIN.....................................................................................................................7
1.2.1 THE HOUSEKEEPING FUNCTION OF THE SMN PROTEIN
.....................................................
7
1.2.2 AXONAL TRANSPORT AND TRANSLATION FUNCTION OF THE SMN PROTEIN
...........................
8
1.2.3 ACTIN REGULATION AND CYTOSKELETAL DYNAMICS FUNCTION OF THE SMN
PROTEIN
.......
8
1.3 SMA ANIMAL
MODELS................................................................................................................9
1.3.1 ZEBRAFISH SMA
MODELS................................................................................................
10
1.3.2 MOUSE SMA M
ODELS....................................................................................................10
1.4 SMA
THERAPY..........................................................................................................................
14
1.4.1 COMPOUNDS CORRECTING ABERRANT SMN2 SPLICING
...................................................
14
1.4.2 COMPOUNDS INCREASING SMN2
TRANSCRIPTION...........................................................16
1.4.3 SMN GENE REPLACEMENT THERAPY USING VIRAL VECTORS
............................................
18
1.4.4 SMN-INDEPENDENT THERAPEUTIC TARGETS (NEUROPROTECTIVE
AGENTS).....................19
1.5 MODIFIER GENES IN SM
A........................................................................................................
20
1.5.1 IDENTIFICATION OF SMA MODIFIERS IN INVERTEBRATE SMA MODELS
...........................
20
1.5.2 IDENTIFICATION OF SMA MODIFIERS IN HUMAN DISCORDANT SMA FAMILIES
................
21
1.5.3 NEUROCALCIN DELTA (NCALD) - A NOVEL HUMAN SMA GENETIC M ODIFIER
................
22
1.5.4 NEURONAL CALCIUM SENSOR 1 (NCS1) AS A POTENTIAL MODIFIER FOR SMA
.................
27
1.6 CAVEOLIN 1 (CAV1) AND ITS POTENTIAL IMPLICATION IN S M A
..............................................
28
1.6.1 ROLE AND FUNCTIONS OF CAVEOLINS
................................................................................
28
1.6.2 CAV1 AND ITS POTENTIAL LINK TO S M A
.........................................................................
29
2. AIMS OF
STUDY................................................................................................................31
3.
RESULTS.............................................................................................................................
35
CHAPTER I: CHARACTERIZATION OF NCALD IN THE BACKGROUND OF S M A
...............
35
3.1 NCALD LOCALIZATION AT RELEVANT SMA PATHOLOGICAL
SITES................................................35
3.1.1 NCALD LOCALIZATION IN PRIMARY M N S
.........................................................................35
3.1.2 NCALD LOCALIZATION AT THE
NMJ..................................................................................
37
3.2 NCALD EXPRESSION DURING NEURONAL DIFFERENTIATION
.....................................................
38
3.3 NCALD DOWNREGULATION IN A SEVERE SMA MOUSE MODEL ON MIXED50
BACKGROUND... 40
3.3.1 GENERATION OF MIXED50 SMA MICE WITH
NCALD HETEROZYGOUS KNOCKOUT...............41
3.3.2 SURVIVAL AND WEIGHT PROGRESSION OF NCALD DOWNREGULATED SMA MICE ON
MIXEDSO
BACKGROUND......................................................................................................................
42
3.3.3 EXPRESSION LEVELS OF SMN AND NCALD IN UNINJECTED MIXED50
BACKGROUND
ANIMALS
.........................................................................................................................................
44
3.3.4 MOTORIC ABILITIES OF NCALD DOWNREGULATED SMA MICE ON MIXED50
BACKGROUND......................................................................................................................................
45
3.4 IMPACT OF NCALD DOWNREGULATION IN LOW-DOSE SMN ASO-INJECTED SMA M
ICE
.......
46
3.4.1 GENERATION OF THE MILD SMA MOUSE MODEL ON MIXED50 BACKGROUND
................
46
3.4.2 FUNCTIONALITY AND DISTRIBUTION OF THE SMN ASO IN THE INJECTED SMA
MICE
......
48
3.4.3 SURVIVAL, GROSS MORPHOLOGY, AND WEIGHT PROGRESSION OF LOW-DOSE SMN
ASO-
INJECTED SMA
MICE..........................................................................................................................49
3.4.4 MOTORIC TESTING OF LOW-DOSE SMN ASO-INJECTED SMA M ICE
................................
52
3.4.5 STRUCTURAL STUDIES OF LOW-DOSE SMN ASO-INJECTED SMA MICE - NMJ AND
MUSCLE FIBER
ANALYSES....................................................................................................................
53
3.4.6 ELECTRONEUROGRAPHIC MEASUREMENTS WITH LOW-DOSE SMN ASO-INJECTED
SMA
MICE
..........................................................................................................................................56
3.4.7 TIME COURSE EXPERIMENTS FOR SMN AND NCALD LEVELS IN SMN
ASO-INJECTED
MIXEDSO BACKGROUND
MICE............................................................................................................
58
3.5 WHY ARE SMA MICE ON MIXEDSO BACKGROUND LESS AFFECTED THAN CONGENIC
C57BL6/N
OR FVB/N SMA
ANIMALS?...................................................................................................................
60
3.6 SMNKO/WT - A MOUSE MODEL FOR MILD
SMA?........................................................................62
CHAPTER II: NCS1 AS A POTENTIAL MODIFIER FOR SPINAL MUSCULAR ATROPHY....
63
3.7 NCS1 LOCALIZATION AND EXPRESSION STUDIES AT RELEVANT SMA PATHOLOGY
SITES
.............
64
3.8 NCS1 AND NEURITE OUTGROWTH REGULATION IN IN VITRO SYSTEMS
........................................
66
3.8.1 NCS1 EXPRESSION DURING NEURONAL DIFFERENTIATION
.................................................
66
3.8.2 ESTABLISHMENT OF NCS1 EXPRESSION REGULATION IN VITRO
.........................................
67
3.8.3 NCS1 DOWNREGULATION TRIGGERS NEURONAL
DIFFERENTIATION......................................68
3.8.4 NCSL DOWNREGULATION RESCUES NEURITE OUTGROWTH IMPAIRMENTS IN SMN-
DEFICIENT NSC34
CELLS.....................................................................................................................
69
3.8.5 NCS1 OVEREXPRESSION IMPAIRS NEURITE OUTGROWTH AND INDUCES MEMBRANE
BLEBBING IN
VITRO.............................................................................................................................
70
3.9 NCSL AND AXON GROWTH REGULATION IN THE SMA ZEBRAFISH MODEL
.................................
72
3.9.1 NCSL DOWNREGULATION INDUCES TERMINAL AXON BRANCHING IN ZEBRAFISH
LARVAE.. 72
3.9.2 NCSL DOWNREGULATION RESCUES AXONAL TRUNCATION PHENOTYPE IN SMA
ZEBRAFISH
LARVAE
.........................................................................................................................................73
3.9.3 NCSL OVEREXPRESSION SHOWS NO AXONAL PHENOTYPE IN ZEBRAFISH LARVAE
...........
74
3.10 POSSIBLE MECHANISTIC ROLE OF NCS1 IN THE SMA PATHOLOGY
...........................................
75
3.10.1 NCSL DOWNREGULATION HAS NO IMPACT ON BULK OR CLATHRIN-MEDIATED
ENDOCYTOSIS......................................................................................................................................75
3.10.2 POSSIBLE INVOLVEMENT OF NCS1 IN THE PERKL/2-DEPENDENT
DIFFERENTIATION
PATHWAY
.........................................................................................................................................77
CHAPTER III: PRELIMINARY DATA ON CAVEOLIN-1 - AN INTERACTION PARTNER OF
S M N
.................................................................................................................................................
79
3.11 EXPRESSION AND LOCALIZATION STUDIES OF CAV1 IN MNS AND AT THE
NMJ.......................80
3.11.1 CAV1 EXPRESSION AND LOCALIZATION STUDIES IN PRIMARY MURINE MNS
...................
80
3.11.2 CAV1 IS EXPRESSED IN THE VASCULATURE IN THE TVA BUT NOT AT THE
NMJ
...............
83
3.12 CAV1 EXPRESSION IN MNS DURING
DIFFERENTIATION.............................................................
84
3.13 EXPRESSION STUDIES OF CAV1 IN WT AND SMA
MEFS.........................................................86
3.14 CAVEOLIN-DEPENDENT ENDOCYTOSIS IN HUMAN
FIBROBLASTS................................................89
4.
DISCUSSION......................................................................................................................
91
4.1 NCALD DOWNREGULATION IN A LOW-DOSE SMN ASO INJECTED SMA MOUSE MODEL
.......
92
4.2 NOVEL POTENTIAL SMA THERAPY TARGETS: NCALD AND PLS3 AS HUMAN SMA
GENETIC
MODIFIERS.............................................................................................................................................
95
4.3 NCALD FUNCTION DURING MN DIFFERENTIATION AND DYNAMIC REGULATION OF
SMN AND
NCALD DURING
AGING...........................................................................................................................
96
4.4 THE USE OF THE SMN ASO-INJECTED MIXED50 MOUSE MODEL AND THE
INFLUENCE OF
GENETIC
BACKGROUND...........................................................................................................................
98
4.5 THE POTENTIAL OF NCS1 AS A NOVEL SMA MODIFIER AND DIRECT COMPARISON
TO
NCALD................................................................................................................................................100
4.6 THE IMPLICATION OF CAV1 IN
SMA......................................................................................
104
5. CONCLUSIONS AND FUTURE PERSPECTIVES
............................................................
107
6. MATERIALS AND
METHODS........................................................................................
109
6.1 M
ATERIALS..............................................................................................................................109
6.1.1 LABORATORY
INSTRUMENTS............................................................................................109
6.1.2 MOUSE WORK
EQUIPMENT...........................................................................................110
6.1.3
CHEMICALS....................................................................................................................I
L L
6.1.4
REAGENTS......................................................................................................................112
6.1.5
KITS................................................................................................................................
113
6.1.6
ENZYMES.......................................................................................................................114
6.1.7 EUKARYOTIC CELL CULTURE
LINES.....................................................................................
114
6.1.8 MOUSE INBRED
STRAINS.................................................................................................115
6.1.9 PRIMARY ANTIBODIES AND STAINING
REAGENTS.............................................................115
6.1.10 SECONDARY
ANTIBODIES................................................................................................116
6.1.11 SOLUTIONS AND M
EDIA..................................................................................................
116
6.1.12
PRIMERS.........................................................................................................................118
6.1.13
SIRNAS...........................................................................................................................
118
6.1.14
MORPHOLINOS................................................................................................................119
6.1.15 GENERATED
PLASMIDS..................................................................................................
119
6.1.16 ANTISENSE OLIGONUCLEOTIDES (ASOS) FOR
IN VIVO
INJECTION.....................................119
6.1.17 SOFTWARE, DATABASES AND INTERNET
PACKAGES........................................................ 120
6.2 M
ETHODS................................................................................................................................121
6.2.1 CELL CULTURE W O RK
........................................................................................................121
6.2.2 NUCLEIC ACID
METHODS................................................................................................125
6.2.3 WORKING WITH
BACTERIA...............................................................................................129
6.2.4 PROTEINBIOCHEMISTRY
METHODS.................................................................................130
6.2.5 HISTOLOGICAL
METHODS.................................................................................................133
6.2.6 WORKING WITH LABORATORY M
ICE................................................................................138
6.2.7 WORKING WITH LABORATORY
ZEBRAFISH........................................................................143
6.2.8 STATISTICAL
ANALYSES....................................................................................................
144
7.
REFERENCES.....................................................................................................................145
8. PUBLICATIONS, PRESENTATIONS, SCHOLARSHIPS AND AW ARDS
....................
168
ACKNOWLEDGMENTS................................................................................................................
XIII
EIDESSTATTLICHE
ERKLAERUNG.................................................................................................
XV
CURRICULUM
VITAE...................................................................................................................XVI
|
| any_adam_object | 1 |
| author | Schneider, Svenja 1986- |
| author_GND | (DE-588)116006492X |
| author_facet | Schneider, Svenja 1986- |
| author_role | aut |
| author_sort | Schneider, Svenja 1986- |
| author_variant | s s ss |
| building | Verbundindex |
| bvnumber | BV045423307 |
| ctrlnum | (OCoLC)1098204779 (DE-599)DNB1161869433 |
| discipline | Biologie |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01421nam a2200349 c 4500</leader><controlfield tag="001">BV045423307</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">t</controlfield><controlfield tag="008">190124s2018 gw a||| m||| 00||| eng d</controlfield><datafield tag="015" ind1=" " ind2=" "><subfield code="a">18,H08</subfield><subfield code="2">dnb</subfield></datafield><datafield tag="016" ind1="7" ind2=" "><subfield code="a">1161869433</subfield><subfield code="2">DE-101</subfield></datafield><datafield tag="020" ind1=" " ind2=" "><subfield code="c">Broschur</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)1098204779</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)DNB1161869433</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rda</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="044" ind1=" " ind2=" "><subfield code="a">gw</subfield><subfield code="c">XA-DE</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-355</subfield><subfield code="a">DE-83</subfield></datafield><datafield tag="084" ind1=" " ind2=" "><subfield code="a">570</subfield><subfield code="2">sdnb</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Schneider, Svenja</subfield><subfield code="d">1986-</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)116006492X</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models</subfield><subfield code="c">vorgelegt von Svenja Schneider aus Oldenburg</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Köln</subfield><subfield code="c">2018</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">170, XVI Seiten</subfield><subfield code="b">Illustrationen</subfield><subfield code="c">21 cm</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="b">Dissertation</subfield><subfield code="c">Universität zu Köln</subfield><subfield code="d">2018</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">B:DE-101</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://d-nb.info/1161869433/04</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030809154&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="999" ind1=" " ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-030809154</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV045423307 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T08:17:45Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-030809154 |
| oclc_num | 1098204779 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-83 |
| owner_facet | DE-355 DE-BY-UBR DE-83 |
| physical | 170, XVI Seiten Illustrationen 21 cm |
| publishDate | 2018 |
| publishDateSearch | 2018 |
| publishDateSort | 2018 |
| record_format | marc |
| spelling | Schneider, Svenja 1986- Verfasser (DE-588)116006492X aut Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models vorgelegt von Svenja Schneider aus Oldenburg Köln 2018 170, XVI Seiten Illustrationen 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Universität zu Köln 2018 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1161869433/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030809154&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schneider, Svenja 1986- Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models |
| subject_GND | (DE-588)4113937-9 |
| title | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models |
| title_auth | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models |
| title_exact_search | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models |
| title_full | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models vorgelegt von Svenja Schneider aus Oldenburg |
| title_fullStr | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models vorgelegt von Svenja Schneider aus Oldenburg |
| title_full_unstemmed | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models vorgelegt von Svenja Schneider aus Oldenburg |
| title_short | Identification and characterization of spinal muscular atrophy (SMA) modifiers - insight from cellular and vertebrate disease models |
| title_sort | identification and characterization of spinal muscular atrophy sma modifiers insight from cellular and vertebrate disease models |
| topic_facet | Hochschulschrift |
| url | http://d-nb.info/1161869433/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030809154&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schneidersvenja identificationandcharacterizationofspinalmuscularatrophysmamodifiersinsightfromcellularandvertebratediseasemodels |
Es ist kein Print-Exemplar vorhanden.
Inhaltsverzeichnis