Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Ulm
2021
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | 89 Blätter Illustrationen, Diagramme 30 cm |
Internformat
MARC
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| 100 | 1 | |a Park, Deung-Dae |e Verfasser |0 (DE-588)1262521882 |4 aut | |
| 245 | 1 | 0 | |a Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |c Deung-Dae Park |
| 264 | 1 | |a Ulm |c 2021 | |
| 300 | |a 89 Blätter |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |b Dissertation |c International Graduate School in Molecular Medicine Ulm |d 2022 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1271066041/04 |3 Inhaltsverzeichnis |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-034560102 | ||
| 883 | 2 | |8 1\p |a dnb |d 20221117 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
Datensatz im Suchindex
| _version_ | 1804185793300791296 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
1.
INTRODUCTION
.....................................................................................................................
10
1.1
CARDIOVASCULAR VASCULAR
DISEASE
(CVD)
.....................................................................
10
1.2
THERAPEUTIC
HEART
FAILURE
STRATEGIES
............................................................................
10
1.3
ZEBRAFISH
AS
A
MODEL
TO
STUDY
HEART
DISEASE
AND
NOVEL
THERAPEUTIC
STRATEGIES
...........
11
1.3.1
FORWARD
AND
REVERSE
GENETIC
APPROACHES
IN
ZEBRAFISH
........................................
12
1.3.2
CARDIOMYOCYTE PROLIFERATION
AND
REGENERATION
IN
ZEBRAFISH
................................
13
1.4
CHROMATIN
REMODELING
AND
THE
SWI/SNF
COMPLEX
...................................................
14
1.4.1
.
THE
STRUCTURE
OF
SWI/SNF
COMPLEX
........................................................
15
1.4.2
SWI/SNF
COMPLEX
FUNCTION
IN
THE
HEART
.............................................................
15
1.5
AIM
OF
WORK
...............................................................................................................
16
2.
MATERIALS
&
METHODS
......................................................................................................
17
2.1
MATERIALS
....................................................................................................................
17
2.1.1
TECHNICAL
INSTRUMENTS
.........................................................................................
17
2.1.2
CONSUMABLES
.......................................................................................................
19
2.1.3
KITS
......................................................................................................................
21
2.1.4
PLASMIDS
.............................................................................................................
22
2.1.5
BACTERIA
STRAINS....................................................................................................
22
2.1.6 RESTRICTION
ENZYMES
............................................................................................
22
2.1.7
CHEMICAL
REAGENTS
...............................................................................................
23
2.1.8
IMPORTANT
COMMERCIAL
ASSAY
...............................................................................
23
2.1.9
ANTIBODIES
..........................................................................................................
24
2.1.10
OLIGONUCLEOTIDES
...............................................................................................
26
2.1.11
FISH
LINES
...........................................................................................................
29
2.1.12
SOFTWARE............................................................................................................
29
2.1.13
DATABASES
..........................................................................................................
29
4
2.2
METHODS
.....................................................................................................................
30
2.2.1
ZEBRAFISH
..............................................................................................................
30
2.2.2
JB-4
EMBEDDING
AND
HEMATOXYLIN
AND
EOSIN
(H&E)
STAINING
...........................
30
2.2.3
MF20/S46
STAINING
..............................................................................................
31
2.2.4
IN
SITU
HYBRIDIZATION
............................................................................................
31
2.2.5
EDU
INCORPORATION
AND
IMMUNOFLUORESCENCE
(IF)
STAINING
.................................
32
2.2.6 IMAGING
AND
COUNTING
CARDIOMYOCYTES
...............................................................
32
2.2.7 POSITIONAL
CLONING
...............................................................................................
33
2.2.8
SEQUENCING GENOMIC
REGION
OF
MUTATION
.............................................................
33
2.2.9
RNA
EXTRACTION
...................................................................................................
33
2.2.10
REVERSE
TRANSCRIPTION
.........................................................................................
34
2.2.11
POLYMERASE
CHAIN REACTION
(PCR)
.....................................................................
34
2.2.12 AGAROSE
GEL
ELECTROPHORESIS
..............................................................................
35
2.2.13
PURIFICATION
AND
ISOLATION
OF
DNA
FROM
AGAROSE
GEL
........................................
35
2.2.14
QUANTITATIVE
REAL
TIME
PCR
(QRT-PCR)
............................................................
35
2.2.15
MICROINJECTION
...................................................................................................
36
2.2.16
CELL
CULTURE
.......................................................................................................
36
2.2.17
SMALL
INTERFERING
RNA
(SIRNA)
KNOCKDOWN
....................................................
37
2.2.18 PROTEIN
EXTRACTION
AND
WESTERN
BLOT
SAMPLE
PREPARATION
(ZEBRAFISH)
...............
37
2.2.19 PROTEIN
EXTRACTION
AND
WESTERN
BLOT
SAMPLE
PREPARATION
(CELL)
.........................
38
2.2.20
WESTERN
BLOT
ANALYSIS
........................................................................................
38
2.2.21
LIBRARY PREPARATION
AND
SEQUENCING..................................................................
38
2.2.22
BIOINFORMATICS
ANALYSIS
OF
DIFFERENTIALLY
EXPRESSED
GENES
................................
39
2.2.23
CRYOINJURY
.........................................................................................................
39
2.2.24
ADULT
ZEBRAFISH
HEART
DISSECTION
AND
ACID
FUCHSIN
ORANG
G
(AFOG)
STAINING
39
2.2.25
CELL
COUNT
AND
VIABILITY
ASSAY
(AC
16)
..............................................................
40
2.2.26
TWO-STEP
CELL
CYCLE
ASSAY
(AC
16)
....................................................................
40
5
2.2.27
STATISTICS............................................................................................................
41
3.
RESULTS
............................................................................................................................
42
3.1
THE
HEART
OF
STONE
MUTATION
AUGMENTS
CARDIOMYOCYTE
PROLIFERATION
IN
ZEBRAFISH
42
3.2
HEART
OF
STONE
ENCODES
ZEBRAFISH
SWI/SNF-RELATED
MATRIX-ASSOCIATED ACTIN
DEPENDENT
REGULATOR
OF
CHROMATIN SUBFAMILY
E
MEMBER
1
(SMARCEL)
..............................
46
3.3
TARGETED
KNOCKDOWN
OF
SMARCEL
IN
WILD-TYPE
ZEBRAFISH
INCREASES
EMBRYONIC
CARDIOMYOCYTE
PROLIFERATION
............................................................................................
48
3.4
ECTOPIC
EXPRESSION
OF
WILD-TYPE
SMARCEL
MRNA
RESCUES
CARDIOMYOCYTE
PROLIFERATION
IN
HOS
MUTANTS
.................................................................................................................
51
3.5
OVEREXPRESSION
OF
SMARCEL
REDUCES
CARDIOMYOCYTE
PROLIFERATION
IN
EMBRYONIC
ZEBRAFISH
.........................................................................................................................
53
3.6
CELL
CYCLE
ACTIVITY
IS
POSITIVELY
REGULATED
BY
THE
LOSS
OF
SMARCEL
FUNCTION
IN
HOS
HEARTS
...............................................................................................................................
56
3.7
SMARCEL
REGULATES
JAK/STAT3
SIGNALING
TO
ORCHESTRATE
CELL
CYCLE
ACTIVITY
IN
ZEBRAFISH
HEARTS
..............................................................................................................................
57
3.8
ECTOPIC
OVEREXPRESSION
OF
STAT
3
MRNA
INDUCES
CARDIOMYOCYTE PROLIFERATION
AND
CARDIAC
HYPERPLASIA
IN
ZEBRAFISH
EMBRYOS.......................................................................
62
3.9
KNOCKDOWN
OF
STAT3
MODERATES
HYPERPLASTIC
CARDIAC
IMPAIRMENT
IN
HOS
MUTANT
.....
63
3.10
SMARCEL
ALSO
INVOLVES
IN
CARDIOMYOCYTE
PROLIFERATION DURING
ADULT
HEART
REGENERATION
IN
ZEBRAFISH
.................................................................................................
65
3.11
THE
ROLE
OF
SMARCEL
IN
CELL
CYCLE
ACTIVATION VIA
THE
JAK/STAT3
PATHWAY
IS
EVOLUTIONARY
CONSERVED....................................................................................................
66
4.
DISCUSSION
.......................................................................................................................
72
5.
SUMMARY
.........................................................................................................................
77
6.
LIST
OF
REFERENCES
...........................................................................................................
78
7.
LIST
OF
PUBLICATIONS
.........................................................................................................
86
8.
ACKNOWLEDGEMENTS.........................................................................................................
87
9.
STATUTORY
DECLARATION
.......................................................................................................
89
6
|
| adam_txt |
TABLE
OF
CONTENTS
1.
INTRODUCTION
.
10
1.1
CARDIOVASCULAR VASCULAR
DISEASE
(CVD)
.
10
1.2
THERAPEUTIC
HEART
FAILURE
STRATEGIES
.
10
1.3
ZEBRAFISH
AS
A
MODEL
TO
STUDY
HEART
DISEASE
AND
NOVEL
THERAPEUTIC
STRATEGIES
.
11
1.3.1
FORWARD
AND
REVERSE
GENETIC
APPROACHES
IN
ZEBRAFISH
.
12
1.3.2
CARDIOMYOCYTE PROLIFERATION
AND
REGENERATION
IN
ZEBRAFISH
.
13
1.4
CHROMATIN
REMODELING
AND
THE
SWI/SNF
COMPLEX
.
14
1.4.1
.
THE
STRUCTURE
OF
SWI/SNF
COMPLEX
.
15
1.4.2
SWI/SNF
COMPLEX
FUNCTION
IN
THE
HEART
.
15
1.5
AIM
OF
WORK
.
16
2.
MATERIALS
&
METHODS
.
17
2.1
MATERIALS
.
17
2.1.1
TECHNICAL
INSTRUMENTS
.
17
2.1.2
CONSUMABLES
.
19
2.1.3
KITS
.
21
2.1.4
PLASMIDS
.
22
2.1.5
BACTERIA
STRAINS.
22
2.1.6 RESTRICTION
ENZYMES
.
22
2.1.7
CHEMICAL
REAGENTS
.
23
2.1.8
IMPORTANT
COMMERCIAL
ASSAY
.
23
2.1.9
ANTIBODIES
.
24
2.1.10
OLIGONUCLEOTIDES
.
26
2.1.11
FISH
LINES
.
29
2.1.12
SOFTWARE.
29
2.1.13
DATABASES
.
29
4
2.2
METHODS
.
30
2.2.1
ZEBRAFISH
.
30
2.2.2
JB-4
EMBEDDING
AND
HEMATOXYLIN
AND
EOSIN
(H&E)
STAINING
.
30
2.2.3
MF20/S46
STAINING
.
31
2.2.4
IN
SITU
HYBRIDIZATION
.
31
2.2.5
EDU
INCORPORATION
AND
IMMUNOFLUORESCENCE
(IF)
STAINING
.
32
2.2.6 IMAGING
AND
COUNTING
CARDIOMYOCYTES
.
32
2.2.7 POSITIONAL
CLONING
.
33
2.2.8
SEQUENCING GENOMIC
REGION
OF
MUTATION
.
33
2.2.9
RNA
EXTRACTION
.
33
2.2.10
REVERSE
TRANSCRIPTION
.
34
2.2.11
POLYMERASE
CHAIN REACTION
(PCR)
.
34
2.2.12 AGAROSE
GEL
ELECTROPHORESIS
.
35
2.2.13
PURIFICATION
AND
ISOLATION
OF
DNA
FROM
AGAROSE
GEL
.
35
2.2.14
QUANTITATIVE
REAL
TIME
PCR
(QRT-PCR)
.
35
2.2.15
MICROINJECTION
.
36
2.2.16
CELL
CULTURE
.
36
2.2.17
SMALL
INTERFERING
RNA
(SIRNA)
KNOCKDOWN
.
37
2.2.18 PROTEIN
EXTRACTION
AND
WESTERN
BLOT
SAMPLE
PREPARATION
(ZEBRAFISH)
.
37
2.2.19 PROTEIN
EXTRACTION
AND
WESTERN
BLOT
SAMPLE
PREPARATION
(CELL)
.
38
2.2.20
WESTERN
BLOT
ANALYSIS
.
38
2.2.21
LIBRARY PREPARATION
AND
SEQUENCING.
38
2.2.22
BIOINFORMATICS
ANALYSIS
OF
DIFFERENTIALLY
EXPRESSED
GENES
.
39
2.2.23
CRYOINJURY
.
39
2.2.24
ADULT
ZEBRAFISH
HEART
DISSECTION
AND
ACID
FUCHSIN
ORANG
G
(AFOG)
STAINING
39
2.2.25
CELL
COUNT
AND
VIABILITY
ASSAY
(AC
16)
.
40
2.2.26
TWO-STEP
CELL
CYCLE
ASSAY
(AC
16)
.
40
5
2.2.27
STATISTICS.
41
3.
RESULTS
.
42
3.1
THE
HEART
OF
STONE
MUTATION
AUGMENTS
CARDIOMYOCYTE
PROLIFERATION
IN
ZEBRAFISH
42
3.2
HEART
OF
STONE
ENCODES
ZEBRAFISH
SWI/SNF-RELATED
MATRIX-ASSOCIATED ACTIN
DEPENDENT
REGULATOR
OF
CHROMATIN SUBFAMILY
E
MEMBER
1
(SMARCEL)
.
46
3.3
TARGETED
KNOCKDOWN
OF
SMARCEL
IN
WILD-TYPE
ZEBRAFISH
INCREASES
EMBRYONIC
CARDIOMYOCYTE
PROLIFERATION
.
48
3.4
ECTOPIC
EXPRESSION
OF
WILD-TYPE
SMARCEL
MRNA
RESCUES
CARDIOMYOCYTE
PROLIFERATION
IN
HOS
MUTANTS
.
51
3.5
OVEREXPRESSION
OF
SMARCEL
REDUCES
CARDIOMYOCYTE
PROLIFERATION
IN
EMBRYONIC
ZEBRAFISH
.
53
3.6
CELL
CYCLE
ACTIVITY
IS
POSITIVELY
REGULATED
BY
THE
LOSS
OF
SMARCEL
FUNCTION
IN
HOS
HEARTS
.
56
3.7
SMARCEL
REGULATES
JAK/STAT3
SIGNALING
TO
ORCHESTRATE
CELL
CYCLE
ACTIVITY
IN
ZEBRAFISH
HEARTS
.
57
3.8
ECTOPIC
OVEREXPRESSION
OF
STAT
3
MRNA
INDUCES
CARDIOMYOCYTE PROLIFERATION
AND
CARDIAC
HYPERPLASIA
IN
ZEBRAFISH
EMBRYOS.
62
3.9
KNOCKDOWN
OF
STAT3
MODERATES
HYPERPLASTIC
CARDIAC
IMPAIRMENT
IN
HOS
MUTANT
.
63
3.10
SMARCEL
ALSO
INVOLVES
IN
CARDIOMYOCYTE
PROLIFERATION DURING
ADULT
HEART
REGENERATION
IN
ZEBRAFISH
.
65
3.11
THE
ROLE
OF
SMARCEL
IN
CELL
CYCLE
ACTIVATION VIA
THE
JAK/STAT3
PATHWAY
IS
EVOLUTIONARY
CONSERVED.
66
4.
DISCUSSION
.
72
5.
SUMMARY
.
77
6.
LIST
OF
REFERENCES
.
78
7.
LIST
OF
PUBLICATIONS
.
86
8.
ACKNOWLEDGEMENTS.
87
9.
STATUTORY
DECLARATION
.
89
6 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Park, Deung-Dae |
| author_GND | (DE-588)1262521882 |
| author_facet | Park, Deung-Dae |
| author_role | aut |
| author_sort | Park, Deung-Dae |
| author_variant | d d p ddp |
| building | Verbundindex |
| bvnumber | BV049298819 |
| ctrlnum | (OCoLC)1335768129 (DE-599)DNB1271066041 |
| discipline | Medizin |
| discipline_str_mv | Medizin |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV049298819 |
| illustrated | Illustrated |
| index_date | 2024-07-03T22:38:43Z |
| indexdate | 2024-07-10T10:00:53Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034560102 |
| oclc_num | 1335768129 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 89 Blätter Illustrationen, Diagramme 30 cm |
| publishDate | 2021 |
| publishDateSearch | 2021 |
| publishDateSort | 2021 |
| record_format | marc |
| spelling | Park, Deung-Dae Verfasser (DE-588)1262521882 aut Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish Deung-Dae Park Ulm 2021 89 Blätter Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation International Graduate School in Molecular Medicine Ulm 2022 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1271066041/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034560102&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p dnb 20221117 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Park, Deung-Dae Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |
| subject_GND | (DE-588)4113937-9 |
| title | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |
| title_auth | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |
| title_exact_search | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |
| title_exact_search_txtP | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |
| title_full | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish Deung-Dae Park |
| title_fullStr | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish Deung-Dae Park |
| title_full_unstemmed | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish Deung-Dae Park |
| title_short | Role of Smarce1, a subunit of the SWI/SNF chromatin remodeling complex, in orchestrating cardiomyocyte proliferation in zebrafish |
| title_sort | role of smarce1 a subunit of the swi snf chromatin remodeling complex in orchestrating cardiomyocyte proliferation in zebrafish |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1271066041/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034560102&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT parkdeungdae roleofsmarce1asubunitoftheswisnfchromatinremodelingcomplexinorchestratingcardiomyocyteproliferationinzebrafish |
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