Role of KIF21A at the glomerular filtration barrier:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Freiburg im Breisgau
[2022]
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | IV, 81 Blätter Illustrationen, Diagramme 30 cm |
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MARC
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| 245 | 1 | 0 | |a Role of KIF21A at the glomerular filtration barrier |c vorgelegt 2021 von Hanna Valentina Riedmann |
| 264 | 1 | |a Freiburg im Breisgau |c [2022] | |
| 300 | |a IV, 81 Blätter |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |b Dissertation |c Albert-Ludwigs-Universität Freiburg im Breisgau |d 2022 | ||
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Record in the Search Index
| _version_ | 1804185911671390208 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
ABBREVIATIONS
.............................................................................................................................
ILL
LIST
OF
TABLES
..............................................................................................................................
IV
LIST
OF
FIGURES
.............................................................................................................................
IV
1.
INTRODUCTION
.....................................................................................................................
1
1.1
THE
HUMAN
KIDNEY
..........................................................................................................
1
1.1.1
FUNCTION
AND
DYSFUNCTION
................................................................................................
1
1.1.2
STRUCTURE
AND
DEVELOPMENTAL
STAGES
..............................................................................
2
1.1.3
RENAL
CORPUSCLES
WITH
GLOMERULAR
FILTRATION
APPARATUS
....................................................
4
1.2
FOCAL
SEGMENTAL
GLOMERULOSCLEROSIS
(FSGS)
..................................................................
6
1.3
ZEBRAFISH
AS
A
MODEL
ORGANISM
......................................................................................
8
1.3.1
PRONEPHROS
.....................................................................................................................
8
1.3.2
DEVELOPMENT
...................................................................................................................
9
1.4
KINESIN
SUPERFAMILY
MOTOR
PROTEINS
(KIFS)
....................................................................
12
1.4.1
KIF21A
(KINESIN
FAMILY
MEMBER
21
A)
...........................................................................
13
1.4.2
KANK
(KIDNEY
ANKYRIN
REPEAT-CONTAINING
PROTEIN)
.......................................................
15
1.5
HYPOTHESIS
AND
EXPERIMENTAL
APPROACH
.......................................................................
16
2.
MATERIAL
AND
METHODS
....................................................................................................
17
2.1
MATERIALS
.......................................................................................................................
17
2.1.1
CHEMICALS
AND
REAGENTS
..............................................................................................
17
2.1.2
BUFFERS
AND
SOLUTIONS
.....................................................................................................
18
2.1.3
BIOLOGICAL
AGENTS
...........................................................................................................
22
2.1.4
ENZYMES
.......................................................................................................................
22
2.1.5
ANTIBODIES
FOR
WESTERN
BLOT
AND
IMMUNOHISTOCHEMISTRY
..............................................
23
2.1.6
PRIMER
...........................................................................................................................
23
2.1.7
MORPHOLINO
ANTISENSE
OLIGONUCLEOTIDES
(MOS)
............................................................
24
2.1.8
ZEBRAFISH
LINES
............................................................................................................
24
2.1.9
BACTERIA
AND
PLASMIDS
.................................................................................................
24
2.1.10
KITS
................................................................................................................................
25
2.1.11
CONSUMABLE
SUPPLIES
...................................................................................................
25
2.1.12
HARDWARE
......................................................................................................................
26
2.1.13
ELECTRONICAL
SUPPORT
......................................................................................................
27
2.2
METHODS
........................................................................................................................
28
2.2.1
IN
VIVO
EXPERIMENTS
......................................................................................................
28
A.
HOUSING
AND
MATING
CONDITIONS
OF
ADULT
ZEBRAFISH
.......................................................
28
B.
EMBRYO
COLLECTION
AND
MAINTENANCE
.............................................................................
28
C.
ZEBRAFISH
MICROINJECTIONS
WITH
MORPHOLINO
ANTISENSE
OLIGONUCLEOTIDES
(MOS)
.............
29
D.
FLUORESCENT
DYE
INJECTION
FILTRATION
ASSAY
......................................................................
32
2.2.2
NUCLEIC
ACID
METHODS
...................................................................................................
33
A.
RNA
ISOLATION
FOR
PREPARING
CDNA
LIBRARY
.....................................................................
33
B.
CDNA
LIBRARY
CONSTRUCTION
.............................................................................................
33
C.
POLYMERASE
CHAIN
REACTION
(PCR)
................................................................................
33
D.
PREPARATION
OF
GENOMIC
ZEBRAFISH
DNA
........................................................................
34
E.
GEL
EXTRACTION
................................................................................................................
34
F.
TRANSFORMATION
OF
ESCHERICHIA
COLI
...............................................................................
34
G.
DNA
PLASMID
PREPARATION
.............................................................................................
35
H.
RESTRICTION
DIGESTION
......................................................................................................
35
I.
DNA
SEQUENCING
...........................................................................................................
36
J.
AGAROSE
GEL
ELECTROPHORESIS
.........................................................................................
36
2.2.3
PROTEIN
METHODS
............................................................................................................
36
A.
PREPARATION
OF
ZEBRAFISH
PROTEIN
LYSATE
.........................................................................
36
B.
SDS-PAGE
...................................................................................................................
36
C.
WESTERN
BLOT
..................................................................................................................
37
D.
PROTEINURIA
ASSAY
..........................................................................................................
37
E.
WHOLE
MOUNT
IN
SITU
HYBRIDIZATION
(WISH)
...................................................................
38
F.
IMMUNOHISTOCHEMISTRY
ON
CRYOSECTIONS
.......................................................................
40
3.
RESULTS
..........................................................................................................................
42
3.1
ZEBRAFISH
KIF21A
PROTEIN
IS
WELL
CONSERVED
COMPARED
WITH
HUMAN
KIF21A
....................
42
3.2
KIF21A
TRANSCRIPTS
ARE
DETECTABLE
DURING
THE
DEVELOPMENT
OF
ZEBRAFISH
.......................
44
3.3
KIF21A
IS
WIDELY
EXPRESSED
IN
ZEBRAFISH
ORGANS
............................................................
45
3.4
WHOLE
MOUNT
IN
SITU
HYBRIDIZATION
(WISH)
DO
NOT
SHOW
SPECIFIC
STAINING
AT
GLOMERULUS45
3.5
IMMUNOSTAINING
ON
CRYOSECTIONS
REVEALS
EXPRESSION
OF
KIF21A
IN
THE
GLOMERULUS
..........
47
3.6
KNOCKDOWN
OF
KIF21A
INDUCES
PERICARDIAL
EDEMA
FORMATION,
IMPLICATING
DISRUPTION
OF
FILTRATION
BARRIER
..............................................................................................................
47
3.7
ZKIF21A
MUTANTS
SHOW
PERICARDIAL
EDEMA
.....................................................................
50
3.8
IMMUNOSTAINING
ON
CRYOSECTIONS
OF
KIF21A
MORPHANTS
AND
MUTANTS
REVEALS
DECREASED
EXPRESSION
SIGNAL
OF
KIF21A
IN
THE
GLOMERULUS
..............................................................
52
3.9
DEXTRAN
PRESENCE
IN
THE
PRONEPHROS
OF
MORPHOLINO
KNOCKDOWN
EMBRYOS
IS
STATISTICALLY
SIGNIFICANT
INCREASED
......................................................................................................
54
3.10
PROTEINURIA
ASSAY
SHOWS
EXCRETION
OF
PROTEIN
IN
MORPHOLINO
KNOCKDOWN
EMBRYOS
AND
MUTANTS
.........................................................................................................................
57
4.
DISCUSSION
.....................................................................................................................
59
5.
SUMMARY
.......................................................................................................................
65
6.
ZUSAMMENFASSUNG
IN
DEUTSCHER
SPRACHE
.....................................................................
66
7.
REFERENCES
....................................................................................................................
67
LEBENSLAUF
................................................................................................................................
77
EIDESSTATTLICHE
VERSICHERUNG
....................................................................................................
79
ERKLARUNG
ZUM
EIGENANTEIL
........................................................................................................
80
DANKSAGUNG
.............................................................................................................................
81
|
| adam_txt |
TABLE
OF
CONTENTS
ABBREVIATIONS
.
ILL
LIST
OF
TABLES
.
IV
LIST
OF
FIGURES
.
IV
1.
INTRODUCTION
.
1
1.1
THE
HUMAN
KIDNEY
.
1
1.1.1
FUNCTION
AND
DYSFUNCTION
.
1
1.1.2
STRUCTURE
AND
DEVELOPMENTAL
STAGES
.
2
1.1.3
RENAL
CORPUSCLES
WITH
GLOMERULAR
FILTRATION
APPARATUS
.
4
1.2
FOCAL
SEGMENTAL
GLOMERULOSCLEROSIS
(FSGS)
.
6
1.3
ZEBRAFISH
AS
A
MODEL
ORGANISM
.
8
1.3.1
PRONEPHROS
.
8
1.3.2
DEVELOPMENT
.
9
1.4
KINESIN
SUPERFAMILY
MOTOR
PROTEINS
(KIFS)
.
12
1.4.1
KIF21A
(KINESIN
FAMILY
MEMBER
21
A)
.
13
1.4.2
KANK
(KIDNEY
ANKYRIN
REPEAT-CONTAINING
PROTEIN)
.
15
1.5
HYPOTHESIS
AND
EXPERIMENTAL
APPROACH
.
16
2.
MATERIAL
AND
METHODS
.
17
2.1
MATERIALS
.
17
2.1.1
CHEMICALS
AND
REAGENTS
.
17
2.1.2
BUFFERS
AND
SOLUTIONS
.
18
2.1.3
BIOLOGICAL
AGENTS
.
22
2.1.4
ENZYMES
.
22
2.1.5
ANTIBODIES
FOR
WESTERN
BLOT
AND
IMMUNOHISTOCHEMISTRY
.
23
2.1.6
PRIMER
.
23
2.1.7
MORPHOLINO
ANTISENSE
OLIGONUCLEOTIDES
(MOS)
.
24
2.1.8
ZEBRAFISH
LINES
.
24
2.1.9
BACTERIA
AND
PLASMIDS
.
24
2.1.10
KITS
.
25
2.1.11
CONSUMABLE
SUPPLIES
.
25
2.1.12
HARDWARE
.
26
2.1.13
ELECTRONICAL
SUPPORT
.
27
2.2
METHODS
.
28
2.2.1
IN
VIVO
EXPERIMENTS
.
28
A.
HOUSING
AND
MATING
CONDITIONS
OF
ADULT
ZEBRAFISH
.
28
B.
EMBRYO
COLLECTION
AND
MAINTENANCE
.
28
C.
ZEBRAFISH
MICROINJECTIONS
WITH
MORPHOLINO
ANTISENSE
OLIGONUCLEOTIDES
(MOS)
.
29
D.
FLUORESCENT
DYE
INJECTION
FILTRATION
ASSAY
.
32
2.2.2
NUCLEIC
ACID
METHODS
.
33
A.
RNA
ISOLATION
FOR
PREPARING
CDNA
LIBRARY
.
33
B.
CDNA
LIBRARY
CONSTRUCTION
.
33
C.
POLYMERASE
CHAIN
REACTION
(PCR)
.
33
D.
PREPARATION
OF
GENOMIC
ZEBRAFISH
DNA
.
34
E.
GEL
EXTRACTION
.
34
F.
TRANSFORMATION
OF
ESCHERICHIA
COLI
.
34
G.
DNA
PLASMID
PREPARATION
.
35
H.
RESTRICTION
DIGESTION
.
35
I.
DNA
SEQUENCING
.
36
J.
AGAROSE
GEL
ELECTROPHORESIS
.
36
2.2.3
PROTEIN
METHODS
.
36
A.
PREPARATION
OF
ZEBRAFISH
PROTEIN
LYSATE
.
36
B.
SDS-PAGE
.
36
C.
WESTERN
BLOT
.
37
D.
PROTEINURIA
ASSAY
.
37
E.
WHOLE
MOUNT
IN
SITU
HYBRIDIZATION
(WISH)
.
38
F.
IMMUNOHISTOCHEMISTRY
ON
CRYOSECTIONS
.
40
3.
RESULTS
.
42
3.1
ZEBRAFISH
KIF21A
PROTEIN
IS
WELL
CONSERVED
COMPARED
WITH
HUMAN
KIF21A
.
42
3.2
KIF21A
TRANSCRIPTS
ARE
DETECTABLE
DURING
THE
DEVELOPMENT
OF
ZEBRAFISH
.
44
3.3
KIF21A
IS
WIDELY
EXPRESSED
IN
ZEBRAFISH
ORGANS
.
45
3.4
WHOLE
MOUNT
IN
SITU
HYBRIDIZATION
(WISH)
DO
NOT
SHOW
SPECIFIC
STAINING
AT
GLOMERULUS45
3.5
IMMUNOSTAINING
ON
CRYOSECTIONS
REVEALS
EXPRESSION
OF
KIF21A
IN
THE
GLOMERULUS
.
47
3.6
KNOCKDOWN
OF
KIF21A
INDUCES
PERICARDIAL
EDEMA
FORMATION,
IMPLICATING
DISRUPTION
OF
FILTRATION
BARRIER
.
47
3.7
ZKIF21A
MUTANTS
SHOW
PERICARDIAL
EDEMA
.
50
3.8
IMMUNOSTAINING
ON
CRYOSECTIONS
OF
KIF21A
MORPHANTS
AND
MUTANTS
REVEALS
DECREASED
EXPRESSION
SIGNAL
OF
KIF21A
IN
THE
GLOMERULUS
.
52
3.9
DEXTRAN
PRESENCE
IN
THE
PRONEPHROS
OF
MORPHOLINO
KNOCKDOWN
EMBRYOS
IS
STATISTICALLY
SIGNIFICANT
INCREASED
.
54
3.10
PROTEINURIA
ASSAY
SHOWS
EXCRETION
OF
PROTEIN
IN
MORPHOLINO
KNOCKDOWN
EMBRYOS
AND
MUTANTS
.
57
4.
DISCUSSION
.
59
5.
SUMMARY
.
65
6.
ZUSAMMENFASSUNG
IN
DEUTSCHER
SPRACHE
.
66
7.
REFERENCES
.
67
LEBENSLAUF
.
77
EIDESSTATTLICHE
VERSICHERUNG
.
79
ERKLARUNG
ZUM
EIGENANTEIL
.
80
DANKSAGUNG
.
81 |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Riedmann, Hanna Valentina |
| author_GND | (DE-588)1261255208 |
| author_facet | Riedmann, Hanna Valentina |
| author_role | aut |
| author_sort | Riedmann, Hanna Valentina |
| author_variant | h v r hv hvr |
| building | Verbundindex |
| bvnumber | BV049367452 |
| ctrlnum | (OCoLC)1333614652 (DE-599)DNB1264252625 |
| 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.BV049367452 |
| illustrated | Illustrated |
| index_date | 2024-07-03T22:53:34Z |
| indexdate | 2024-07-10T10:02:45Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034627479 |
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| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | IV, 81 Blätter Illustrationen, Diagramme 30 cm |
| publishDate | 2022 |
| publishDateSearch | 2022 |
| publishDateSort | 2022 |
| record_format | marc |
| spelling | Riedmann, Hanna Valentina Verfasser (DE-588)1261255208 aut Role of KIF21A at the glomerular filtration barrier vorgelegt 2021 von Hanna Valentina Riedmann Freiburg im Breisgau [2022] IV, 81 Blätter Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Albert-Ludwigs-Universität Freiburg im Breisgau 2022 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1264252625/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034627479&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p dnb 20220825 DE-101 https://d-nb.info/provenance/plan#dnb 2\p dnb 20220825 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Riedmann, Hanna Valentina Role of KIF21A at the glomerular filtration barrier |
| subject_GND | (DE-588)4113937-9 |
| title | Role of KIF21A at the glomerular filtration barrier |
| title_auth | Role of KIF21A at the glomerular filtration barrier |
| title_exact_search | Role of KIF21A at the glomerular filtration barrier |
| title_exact_search_txtP | Role of KIF21A at the glomerular filtration barrier |
| title_full | Role of KIF21A at the glomerular filtration barrier vorgelegt 2021 von Hanna Valentina Riedmann |
| title_fullStr | Role of KIF21A at the glomerular filtration barrier vorgelegt 2021 von Hanna Valentina Riedmann |
| title_full_unstemmed | Role of KIF21A at the glomerular filtration barrier vorgelegt 2021 von Hanna Valentina Riedmann |
| title_short | Role of KIF21A at the glomerular filtration barrier |
| title_sort | role of kif21a at the glomerular filtration barrier |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1264252625/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034627479&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT riedmannhannavalentina roleofkif21aattheglomerularfiltrationbarrier |
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