Single cell migration in live embryos: the role of E-cadherin in zebrafish germ cell migration
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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Münster
2019
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| Beschreibung: | 96, VIII Blätter Illustrationen 30 cm 1 CD-ROM |
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MARC
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| 100 | 1 | |a Schumacher, Isabel |d 1992- |e Verfasser |0 (DE-588)1190048574 |4 aut | |
| 245 | 1 | 0 | |a Single cell migration in live embryos |b the role of E-cadherin in zebrafish germ cell migration |c vorgelegt von Schumacher, Isabel |
| 264 | 1 | |a Münster |c 2019 | |
| 300 | |a 96, VIII Blätter |b Illustrationen |c 30 cm |e 1 CD-ROM | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Westfälische Wilhelms-Universität Münster |d 2019 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-031420554 | ||
Datensatz im Suchindex
| _version_ | 1804180295680786432 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
1
INTRODUCTION
...........................................................................................................
1
1.1
SINGLE
CELL
MIGRATION
..................................................................................................
1
1.1.1
CLASSICAL
MIGRATION
MODES
....................................................................................
1
1.1.2
FORCE
TRANSMISSION
IN
ADHESION-INDEPENDENT
MIGRATION
.................................
2
1.2
ZEBRAFISH
PGCS
AS
MODEL
FOR
SINGLE
CELL
MIGRATION
..............................................
4
1.2.1
OVERVIEW
OF
ZEBRAFISH
PGC
MIGRATION
.................................................................
4
1.2.2
ZEBRAFISH
PGCS
EMPLOY
BLEBBING
MOTILITY
...........................................................
5
1.2.3
THE
MOLECULAR
BASIS
FOR
ZEBRAFISH
PGC
MIGRATION
...............................................
6
1.3
THE
CELL-CELL
ADHESION
PROTEIN
E-CADHERIN
..........................................................
8
1.3.1
E-CADHERIN
CONNECTS
THE
EXTRACELLULAR
ENVIRONMENT
WITH
THE
INTRACELLULAR
CYTOSKELETON
......................................................................................................................
8
1.3.2
THE
ROLE
OF
CADHERINS
IN
CELL
MIGRATION
..............................................................
9
1.4
AIM
OF
THIS
STUDY
.....................................................................................................
11
2
MATERIAL
AND
METHODS
........................................................................................
12
2.1
MATERIALS
...................................................................................................................
12
2.1.1
ZEBRAFISH
STRAINS
..................................................................................................
12
2.1.2
BACTERIA
FOR
PLASMID
PROPAGATION
......................................................................
12
2.1.3
COMMERCIAL
KITS
..................................................................................................
12
2.1.4
REAGENTS,
CHEMICALS,
MEDIA
AND
BUFFER
.............................................................
13
2.1.5
CONSTRUCTS
............................................................................................................
17
2.1.6
MORPHOLINOS
........................................................................................................
20
2.1.7
ANTISENSE
OLIGONUCLEOTIDES
................................................................................
20
2.1.8
SHRNA
SEQUENCES
................................................................................................
21
2.1.9
ANTIBODIES
............................................................................................................
22
2.1.10
TECHNICAL
EQUIPMENT
.......................................................................................
22
2.1.11
SOFTWARE
............................................................................................................
24
2.2
METHODS
....................................................................................................................
25
2.2.1
FISH
MAINTENANCE
AND
EMBRYO
STAGING
.............................................................
25
2.2.2
INJECTION
PROCEDURE
............................................................................................
26
2.2.3
GENERATION
OF
A
CXCLL2A
KD
BACKGROUND
............................................................
28
2.2.4
CLONING
PROTOCOLS
................................................................................................
28
2.2.5
GENERATION
OF
TRANSGENIC
FISH
LINES..................................................................
32
2.2.6
FIXATION
OF
EMBRYOS
...........................................................................................
33
2.2.7
WHOLE-MOUNT
IN
SITU
HYBRIDIZATION
(WMISH)
..................................................
33
2.2.8
ANTIBODY
STAINING
................................................................................................34
2.2.9
EMBRYO
PREPARATION
FOR
IMAGING
.......................................................................
35
2.2.10
IMAGE
ACQUISITION
.............................................................................................
35
2.2.11
ANALYSIS
OF
IMAGING
DATA
..................................................................................
36
2.2.12
STATISTICAL
ANALYSIS
...........................................................................................
37
3
RESULTS
.................................................................................................................
38
3.1
GENERATION
OF
E-CADHERIN
DEPLETED
CLONES
.........................................................
33
3.1.1
A
FLUORESCENTLY
LABELED
MO
EFFICIENTLY
KNOCKS
DOWN
E-CADHERIN
.................
38
3.1.2
32-CELL
INJECTION
LEADS
TO
APPROPRIATE
CLONE
SIZE
........................................... 41
3.2
THE
E-CADHERIN
LEVEL
OF
THE
ENVIRONMENT
AFFECTS
WT
PGC
MIGRATION
AND
POLARITY
...............................................................................................................................
45
3.2.1
THE
MIGRATORY
PATH
OF
WT
PGCS
IS
ALTERED
BY
AN
E-CADHERIN
DEPLETED
ENVIRONMENT
..................................................................................................................
45
3.2.2
MO-RESISTANT
E-CADHERIN
RNA
RESTORES
NO
REACTION
UPON
TOUCHING
THE
TREATED
CLONE
.................................................................................................................49
3.2.3
THE
DIFFERENCE
IN
REACTING
TO
A
TREATED
CLONE
IS
INDEPENDENT
OF
THE
PRESENCE
OF
THE
CHEMOKINE
CXCLL2A
...............................................................................................
50
3.3
CONSTANT
CONTACT
TO
E-CADHERIN
KD
CLONE
AFFECTS
THE
POLARITY
BUT
NOT
THE
MIGRATION
SPEED
OF
WT
PGCS
...........................................................................................52
3.4
E-CADHERIN
KD
PGCS
ARE
MOTILE
IN
A
WT
ENVIRONMENT
.....................................
56
3.5
EMBRYOS
WITH
E-CADHERIN
KD
CLONE
HAVE
MORE
ECTOPIC
PGCS
AT
24
HPF
......
59
3.6
ESTABLISHMENT
OF
A
KD
OF
E-CADHERIN
IN
PGCS
VIA
AN
SHRNA
............................
60
3.6.1
ASSESSMENT
OF
E-CADHERIN
SHRNA
TARGET
SEQUENCES
.........................................
60
3.6.2
GLOBAL
INDUCTION
OF
THE
TRANSGENIC
UAS:SHRNA3
E
CADHERIN
CAN
INDUCE
A
REDUCTION
OF
E-CADHERIN
EXPRESSION
................................................................................................64
3.6.3
THE
EXPRESSION
OF
GAL4FF
IN
THE
PGCS
LEADS
TO
A
TISSUE
SPECIFIC
ACTIVATION
OF
GENES
DOWNSTREAM
OF
AN
UAS
.......................................................................................
71
4
DISCUSSION
...........................................................................................................
75
4.1
E-CADHERIN
LEVEL
OF
ENVIRONMENT
INFLUENCES
PGC
MIGRATION
AND
POLARITY
...
75
4.2
E-CADHERIN
IS
DISPENSABLE
FOR
PGC
MOTILITY
........................................................
80
4.3
EVALUATION
OF
PGC
SPECIFIC
KD
OF
E-CADHERIN
USING
A
TRANSGENIC
UAS:SHRNA
E
CADHERLN
FISH
LINE
.............................................................................................
82
5
SUMMARY
..............................................................................................................
85
6
REFERENCES
...........................................................................................................
86
7
ACKNOWLEDGEMENTS
............................................................................................
94
8
CURRICULUM
VITAE
................................................................................................
95
9
APPENDIX
.................................................................................................................
I
9.1
ABBREVIATIONS
.............................................................................................................
I
9.2
LIST
OF
FIGURES
............................................................................................................
IV
9.3
LIST
OF
TABLES
.............................................................................................................
VI
9.4
LIST
OF
MOVIES
..........................................................................................................
VII
9.5
STATISTICS
OF
BEHAVIOR
UPON
CONTACTING
A
MO
TREATED CLONE
...........................
VII
|
| any_adam_object | 1 |
| author | Schumacher, Isabel 1992- |
| author_GND | (DE-588)1190048574 |
| author_facet | Schumacher, Isabel 1992- |
| author_role | aut |
| author_sort | Schumacher, Isabel 1992- |
| author_variant | i s is |
| building | Verbundindex |
| bvnumber | BV046038819 |
| ctrlnum | (OCoLC)1108158756 (DE-599)DNB1189615363 |
| discipline | Biologie Medizin |
| format | Thesis Book |
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| indexdate | 2024-07-10T08:33:30Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-031420554 |
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| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | 96, VIII Blätter Illustrationen 30 cm 1 CD-ROM |
| publishDate | 2019 |
| publishDateSearch | 2019 |
| publishDateSort | 2019 |
| record_format | marc |
| spelling | Schumacher, Isabel 1992- Verfasser (DE-588)1190048574 aut Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration vorgelegt von Schumacher, Isabel Münster 2019 96, VIII Blätter Illustrationen 30 cm 1 CD-ROM txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische Wilhelms-Universität Münster 2019 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1189615363/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031420554&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Schumacher, Isabel 1992- Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration |
| subject_GND | (DE-588)4113937-9 |
| title | Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration |
| title_auth | Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration |
| title_exact_search | Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration |
| title_full | Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration vorgelegt von Schumacher, Isabel |
| title_fullStr | Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration vorgelegt von Schumacher, Isabel |
| title_full_unstemmed | Single cell migration in live embryos the role of E-cadherin in zebrafish germ cell migration vorgelegt von Schumacher, Isabel |
| title_short | Single cell migration in live embryos |
| title_sort | single cell migration in live embryos the role of e cadherin in zebrafish germ cell migration |
| title_sub | the role of E-cadherin in zebrafish germ cell migration |
| topic_facet | Hochschulschrift |
| url | http://d-nb.info/1189615363/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031420554&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schumacherisabel singlecellmigrationinliveembryostheroleofecadherininzebrafishgermcellmigration |
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