The role of actin filament dynamics in cell motility:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
1999
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | V, 165 S. Ill., graph. Darst. |
Internformat
MARC
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| 001 | BV012881453 | ||
| 003 | DE-604 | ||
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| 007 | t | ||
| 008 | 991130s1999 ad|| m||| 00||| eng d | ||
| 016 | 7 | |a 958406987 |2 DE-101 | |
| 035 | |a (OCoLC)247354992 | ||
| 035 | |a (DE-599)BVBBV012881453 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-19 |a DE-12 |a DE-29T |a DE-355 |a DE-188 | ||
| 100 | 1 | |a Hangen-Mordi, Ifeaka |d 1966- |e Verfasser |0 (DE-588)121781593 |4 aut | |
| 245 | 1 | 0 | |a The role of actin filament dynamics in cell motility |c Ifeaka Hangen-Mordi |
| 264 | 1 | |c 1999 | |
| 300 | |a V, 165 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Univ., Diss., 1999 | ||
| 650 | 0 | 7 | |a Motilität |0 (DE-588)4114638-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Dynamik |0 (DE-588)4013384-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Actin-Filament |0 (DE-588)4211337-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Zelle |0 (DE-588)4067537-3 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a Zelle |0 (DE-588)4067537-3 |D s |
| 689 | 0 | 1 | |a Motilität |0 (DE-588)4114638-4 |D s |
| 689 | 0 | 2 | |a Actin-Filament |0 (DE-588)4211337-4 |D s |
| 689 | 0 | 3 | |a Dynamik |0 (DE-588)4013384-9 |D s |
| 689 | 0 | |5 DE-604 | |
| 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=008768197&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-008768197 | |
Datensatz im Suchindex
| _version_ | 1807593761734131712 |
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| adam_text |
1
1NTRODUCTION
1
1.1
MOTIVATION
AND
AIM
I
1.2
APPROACH
3
1.3
BACKGROUND
5
1.3.1
THE
EUKARYOTIC
CYTOSKELETON
5
1
.3.2
MICROFILAMENT
ORGANISATION
AND
REGULATION
6
1.
3.2.
1
FILAMENTOUS
ACTIN
BINDING
PROTEINS
7
1.3.2.2
ACTIN
MONOMER
BINDING
(SEQUESTERING/DEPOLYMERISING)
PROTEINS
9
1
.3.3
MICROFILAMENT
STRUCTURES
IN
CELL
LOCOMOTION
9
1.3.3.
1
MICROFILAMENT
STRUCTURES
IN
THE
CELL
9
1.3.3.2
PROCESSES
INVOLVED
IN
CELL
LOCOMOTION
10
1.3.4
SIGNALLING
TO
THE
ACTIN
CYTOSKELETON
12
1.3.4.1
THE
SMALL
GTP-BINDING
PROTEINS
RHO,
RAC
AND
CDC42
13
1.3.4.2
POST-TRANSLATIONAL
MODIFICATION
OF
RHOA,
RAC
AND
CDC42
14
1.3.5
CYTOCHALASINS
15
2
MATERIALS
AND
METHODS
18
2.1
MATERIALS
18
2.1.1
REAGENTS
18
2.1.2
CELL
CULTURE
REAGENTS
AND
MEDIA
20
2.1.3
COLUMN
MATERIALS
20
2.1.4
OTHER
MATERIALS
21
2.1.5
EQUIPMENT
21
2.1.5.1
MICROSCOPES
AND
PHOTOMETERS
2
1
2.1.5.2
CAMERAS,
COMPUTERS
AND
ACCESSORIES
22
2.
1.
5.3
CENTRIFUGES
22
2.1.5.4
OTHER
EQUIPMENT
23
2.1.6
ANTIBODIES
23
2.1.6.1
PRIMARY
ANTIBODIES
23
2.1.6.2
SECONDARY
ANTIBODIES
24
2.1.7
RADIOCHEMICALS
24
2.1.8
CELL
LINES
24
2.1.9
STOCK
SOLUTIONS
24
2.1.10
BUFFERS
25
2.2
MICROSCOPY
METHODS
26
2.2.1
INDIRECT
IMMUNOFLUORESCENCE
26
2.2.2
QUICK
PERMEABILISATION/FIXATION
PROCEDURE
OF
CELLS
UNDER
MICROSCOPE OBSERVATION
27
2.2.3
IN
VITRO
MOTILITY
OF
ACTIN
FILAMENTS
ON
HEAVY
MEROMYOSIN
28
2.2.4
MICROSCOPE
EXPERIMENTS
WITH
LIVING
CELLS
-
GENERAL
PROCEDURES
29
2.2.5
MICROSCOPE
SETUP
AND
IMAGE
AQUISITION
30
2.2.6
IMAGE
ANALYSIS
AND
STATISTICAL
PROCEDURES
33
2.3
CELL
BIOLOGY
METHODS
33
2.3.1
CELL
CULTURE
33
2.3.1.1
PASSAGING
AND
SEEDING
OF
CELLS
34
2.3.
1.
1.1
CLEANING
OF
COVERSLIPS
FOR
COATING
AND
SEEDING
'
34
2.3.
1
.1.2
POLY-L-LYSINE
COATING
OF
COVERSLIPS
34
2.3.
1
.2
FREEZE-STORAGE
OF
CELLS
34
2.3.1.3
LABELLING
OF
CELLS
IN
VIVO
WITH
2-'*C-MEVALONOLACTONE
35
2.3.2
MICROINJECTION
OF
CELLS
35
2.4
PROTEIN
CHEMISTRY
METHODS
36
2.4.
1
PREPARATION
OF
ACETONE-EXTRACTED
POWDER
FROM
RABBIT
MUSCLE
36
2.4.2
PURIFICATION
OF
ACTIN
FROM
ACETONE-EXTRACTED
POWDER
37
2.4.3
PURIFICATION
OF
MYOSIN
II
FROM
RABBIT
MUSCLE
37
2.4.4
CHYMOTRYPTIC
CLEAVAGE
OF
MYOSIN
TO
HEAVY
MEROMYOSIN
(HMM)
38
2.4.5
PURIFICATION
OF
A-ACTININ
38
2.4.6
DETERMINATION
OF
PROTEIN
CONCENTRATION
39
2.4.7
SPIN-DOWN
ASSAY
39
2.4.8
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
AND WESTERN
BLOTTING
40
2.4.8.1
PREPARATION
OF
TOTAL
CELL
LYSATE
4
1
2.4.8.2
MOLECULAR
WEIGHT
STANDARDS
41
2.4.9
TWO-DIMENSIONAL
(2-D)
GEL
ELECTROPHORESIS
41
2.4.9.
1
SAMPLE
PREPARATION
FOR
2-D-ELECTROPHORESIS
42
2.4.10
COOMASSIE-BLUE-STAINING
OF
PROTEINS
43
2.4.11
PONCEAU-S
STAINING
OF
BLOTTED
PROTEINS
43
2.4.12
IMMUNODETECTION
OF
BLOTTED
PROTEINS
43
2.4.13
STRIPPING
OF
PROTEIN
BLOTS
44
2.4.14
TCA
PRECIPITATION
OF
PROTEINS
44
2.4.15
CONJUGATION
OF
PROTEINS
TO
FLUORESCENT
DYES
44
3
RESULTS
46
3.1
ACTIN
FILAMENT
DYNAMICS
DURING
THE
REFLATTENING
OF
LOVASTATIN-ROUNDED
CELLS
46
3.1.1
MORPHOLOGY
AND
MICROFILAMENT
STRUCTURE
OF
A
FIBROBLAST
46
3.1.2
CHARACTERISATION
OF
LOVASTATIN-ROUNDED
CELLS
47
3.1.2.
1
MORPHOLOGY
AND
MOVEMENTS
OF
LOVASTATIN-ROUNDED
CELLS
48
3.1.2.2
CHARACTERISTICS
OF
MEVALONATE-INITIATED
REFLATTENING
OF
LOVASTATIN-TREATED
ROUNDED
CELLS
49
3.1.3
LOCALISATION
OF
F-ACTIN
AND
A-ACTININ
IN
REFLATTENING
CELLS
50
3.1.4
FORMATION
OF
FOCAL
CONTACTS
IN
REFLATTENING
CELLS
50
3.1.5
THE
RESTRUCTURING
OF
THE
ACTIN
CYTOSKELETON
DURING
REFLATTENING:
OBSERVATIONS
IN
LIVING
CELLS
56
3.
1
.5.
1
FLUORESCENT
PHALLOIDIN
IS
A
SUITABLE
PROBE
FOR
THE
DYNAMICS
OF
THE
ACTIN
CYTOSKELETON
IN
LIVING
CELLS56
3.1.5.2
THE
DISRUPTED
ACTIN
CYTOSKELETON
OF
LOVASTATIN-TREATED
CELLS
RETAINS
A
DISTINCT
STRUCTURE
OF
RESIDUAL
FILAMENTS
59
3.1.5.3
THE
PROCESS
OF
REFLATTENING
-
MORPHOLOGICAL
AND
CYTOSKELETAL
CHANGES
62
3.1.5.4
FORWARD-DIRECTED
MOTILE
AND
IMMOTILE
ACTIN
BUNDLE
POPULATIONS
ARE
PRESENT
IN
THE
CELL
BODY
OF
REFLATTENING
CELLS
67
3.
1.
5.5
THE
ACTIN-BASED
PROCESS
OF
REFLATTENING
OF
LOVASTATIN-TREATED,
ROUNDED
CELLS
IS
COMPLETELY
INHIBITED
IN
THE
PRESENCE
OF
CYTOCHALASIN
B
(CB)
AND
IN
THE
ABSENCE
OF
EXOGENOUS
MG
2
'
AND
CA
2
*
72
3.1.6
G-ACTIN
TO
F-ACTIN
RATIO
IS
NOT
ALTERED
IN
THE
LOVASTATIN-TREATED,
ROUNDED
CELLS
75
3.1.7
THE
ROLE
OF
MYOSIN
11
IN
THE
REFLATTENING
PROCESS
76
3.1.7.1
LOVASTATIN-ROUNDED
CELLS
ARE
ABLE
TO
REFLATTEN
IN THE
PRESENCE
OF
A
MYOSIN
II
INHIBITOR
77
3.1.7.2
THE
LOCALISATION
OF
MYOSIN
II
IN
REFLATTENING
CELLS
81
3.2
THE
SMALL
GTP-BINDING
PROTEIN
RHOA
PLAYS
A
MAJOR
ROLE
IN
THE
REFLATTENING
OF
LOVASTATIN-TREATED
CELLS84
3.2.1
EVIDENCE
FOR
RHOA
PLAYING
A
MAJOR
ROLE
IN
REFLATTENING
AND
THE
REORGANISATION
OF
ACTIN
IN
LOVASTATIN
TREATED
CELLS
84
3.2.1.1
RHOA
IS
ONE
OF
THE
FIRST
PROTEINS
LABELLED
IN
VIVO
BY
RADIOACTIVE
MEVALONATE
84
3.2.1.2
LPA,
WHICH
ACTIVATES
A
SPECIFICALLY
RHOA-DEPENDENT
PATHWAY,
CAN
INDUCE
REFLATTENING
OF
LOVASTATIN
TREATED
ROUNDED
CELLS
90
3.2.2
RHOA
IS
NOT
PREDOMINANTLY
LOCALISED
AT
THE
MEMBRANE
DURING
THE
REFLATTENING
OF
LOVASTATIN-ROUNDED
CELLS9L
3.2.3
SERUM-DEPENDENT
REFLATTENING
MAY
BE
INDUCED
INDEPENDENTLY
OF
MEVALONATE
93
3.2.3.1
LOVASTATIN-TREATED
ROUNDED
CELLS
REFLATTEN
UPON
MICROINJECTION
OF
THE
CDNA
OF
CONSTITUTIVELY
ACTIVE
RHOA
95
3.2.4
LOVASTATIN-ROUNDED
CELLS
EXHIBIT
AN
ALTERED
PHOSPHORYLATION
PATTERN
AND
DO
NOT
REFLATTEN
IN
THE
PRESENCE
OF
TYROSINE
PHOSPHORYLATION
INHIBITORS
96
3.3
CYTOCHALASIN
B
-
A
PROBE
FOR
DISTINGUISHING
BETWEEN
FUNCTIONALLY
DIVERSE
F-ACTIN
STRUCTURES
100
3.3.1
HIGH
CONCENTRATIONS
OF
CB
REVEAL
SUBPOPULATIONS
OF
ACTIN
FIBRES
101
3.3.2
SEQUENTIAL
REFORMATION
OF
THE
CYTOSKELETON
UPON
REMOVAL
OF
CB
106
3.3.3
EFFECT
OF
LOW
CONCENTRATIONS
OFCB
ON
WOUND-HEALING
FIBROBLASTS:
100
NM
IS
THE
'
THRESHOLD
CONCENTRATION
'
FOR
THE
DEVELOPMENT
OF
LAMELLIPODIA
1
09
3.3.4
EFFECT
OF
LOW
CONCENTRATIONS
OF
CB
ON
WOUND-HEALING
FIBROBLASTS:
WOUND-HEALING
CAN
TAKE
PLACE
IN
THE
ABSENCE
OF
LAMELLIPODIAL
PROTRUSION
BUT
IS
RETARDED
BY
A
MYOSIN
II
INHIBITOR
1
15
3.3.4.
1
WOUND-HEALING
CAN
TAKE
PLACE
IN
THE
ABSENCE
OF
LAMELLIPODIAL
PROTRUSION
1
15
3.3.4.2
WOUND-HEALING
IS
RETARDED
BY
A
MYOSIN
II
INHIBITOR
1
18
3.3.5
NEW
FILAMENT
FORMATION
IS
NOT
ESSENTIAL
FOR
CYTOKINESIS
1
20
3.3.6
CYTOKINESIS
CAN
OCCUR
IN
THE
ABSENCE
OF
FUNCTIONAL
MYOSIN
II
122
3.3.7
THE
INHIBITION
OF
MYOSIN
II
BY
BDM
TESTED
IN
AN
IN
VITRO
MOTILITY
ASSAY
125
4
DISCUSSION
128
4.1
ACTIN
FILAMENT
DYNAMICS
DURING
REFIATTENING
OF
LOVASTATIN-ROUNDED
CELLS:
A
MODEL
SYSTEM
FOR
CELL
SPREADING
AND
OTHER
ACTIN-BASED
PROCESSES
128
4.1.1
LOVASTATIN-ROUNDED
CELLS
-
A
NOVEL
LIVING
CELL
SYSTEM
FOR
STUDYING
ACTIN
DYNAMICS
1
28
4.1.2
MORPHOLOGY
AND
SPREADING
MECHANISMS
DURING
REVERSION
129
4.
1
.3
TEMPORAL
DISCRIMINATION
IN
ACTIN-BASED
PROCESSES
DURING
THE
RESTRUCTURING
OF
THE
ACTIN
MICROFILAMENT
SYSTEM
1
32
4.
1
.4
FORWARD-MOVING
ACTIN
FILAMENTS,
A
PROCESS
COMMON
TO
GENERAL
CELL
MOTILITY?
133
4.1.5
PHALLOIDIN
SELECTIVELY
LABELS
ACTIN
FILAMENTS
IN
THE
LAMELLIPODIUM
1
35
4.1.6
THE
G
TO
F-ACTIN
RATIO
IS
APPARENTLY
UNPERTURBED
IN
LOVASTATIN-TREATED
ROUNDED
CELLS
AND
THIS
MAY
BE
DUE
TO
THE
INFLUENCE
OF
REMNANT
FILAMENTOUS
ACTIN
136
4.2
REFIATTENING
OF
CELLS
AND
REFORMATION
OF
THE
ACTIN
CYTOSKELETON
IS
MEDIATED
VIA
PRENYLATED
AND
ACTIVATED
RHOA
137
4.2.1
RHOA
MIGHT
BE
PHOSPHORYLATED
DEPENDING
ON
THE
DEGREE
OF
PRENYLATION
1
37
4.2.2
ACTIVATED
RHO
DOES
NOT
NEED
PRENYLATION
FOR
FUNCTION
139
4.2.3
THE
PROCESS
OF
REFIATTENING
REQUIRES
THE
TYROSINE
PHOSPHORYLATION
OF
PROTEIN(S)
UPSTREAM
OF
RHOA
141
4.3
DISTINCT
ACTIN
STRUCTURES
AND
THEIR
FUNCTIONS
CAN
BE
PROBED
BY
USING
CB
OVER
A
BROAD
CONCENTRATION
RANGE
143
4.3.1
FUNCTIONAL
DISCRIMINATION
BETWEEN
ACTIN-BUNDLE
(STRESS
FIBRE)
SUB-POPULATIONS
BY
HIGH
CONCENTRATIONS
OF
CB
143
4.3.2
CB
AT
100
NM
IS
THE
'
THRESHOLD
CONCENTRATION'
FOR
THE
COLLAPSE
OF
LAMELLIPODIA
AND
CAPPING
OF
LAMELLIPODIAL
FILAMENTS
147
4.3.3
WOUND-HEALING
CAN
OCCUR
IN
THE
ABSENCE
OF
LAMELLIPODIA
BUT
REQUIRES
MYOSIN
148
IV
4.3.4
THE
CLEAVAGE
FURROW
IN
CYTOKINESIS
IS
FORMED
FROM
PRE-ASSEMBLED
ACTIN
FILAMENTS
4.3.5
MYOSIN
II
IS
NOT
ESSENTIAL
FOR
CYTOKINESIS
5
SUMMARY
149
150
152
6
LITERATURE
155
ABBREVIATIONS
ACKNOWLEDGEMENTS
CURRICULUM
VITAE
V |
| any_adam_object | 1 |
| author | Hangen-Mordi, Ifeaka 1966- |
| author_GND | (DE-588)121781593 |
| author_facet | Hangen-Mordi, Ifeaka 1966- |
| author_role | aut |
| author_sort | Hangen-Mordi, Ifeaka 1966- |
| author_variant | i h m ihm |
| building | Verbundindex |
| bvnumber | BV012881453 |
| ctrlnum | (OCoLC)247354992 (DE-599)BVBBV012881453 |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV012881453 |
| illustrated | Illustrated |
| indexdate | 2024-08-17T00:49:02Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008768197 |
| oclc_num | 247354992 |
| open_access_boolean | |
| owner | DE-19 DE-BY-UBM DE-12 DE-29T DE-355 DE-BY-UBR DE-188 |
| owner_facet | DE-19 DE-BY-UBM DE-12 DE-29T DE-355 DE-BY-UBR DE-188 |
| physical | V, 165 S. Ill., graph. Darst. |
| publishDate | 1999 |
| publishDateSearch | 1999 |
| publishDateSort | 1999 |
| record_format | marc |
| spelling | Hangen-Mordi, Ifeaka 1966- Verfasser (DE-588)121781593 aut The role of actin filament dynamics in cell motility Ifeaka Hangen-Mordi 1999 V, 165 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Univ., Diss., 1999 Motilität (DE-588)4114638-4 gnd rswk-swf Dynamik (DE-588)4013384-9 gnd rswk-swf Actin-Filament (DE-588)4211337-4 gnd rswk-swf Zelle (DE-588)4067537-3 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Zelle (DE-588)4067537-3 s Motilität (DE-588)4114638-4 s Actin-Filament (DE-588)4211337-4 s Dynamik (DE-588)4013384-9 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008768197&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Hangen-Mordi, Ifeaka 1966- The role of actin filament dynamics in cell motility Motilität (DE-588)4114638-4 gnd Dynamik (DE-588)4013384-9 gnd Actin-Filament (DE-588)4211337-4 gnd Zelle (DE-588)4067537-3 gnd |
| subject_GND | (DE-588)4114638-4 (DE-588)4013384-9 (DE-588)4211337-4 (DE-588)4067537-3 (DE-588)4113937-9 |
| title | The role of actin filament dynamics in cell motility |
| title_auth | The role of actin filament dynamics in cell motility |
| title_exact_search | The role of actin filament dynamics in cell motility |
| title_full | The role of actin filament dynamics in cell motility Ifeaka Hangen-Mordi |
| title_fullStr | The role of actin filament dynamics in cell motility Ifeaka Hangen-Mordi |
| title_full_unstemmed | The role of actin filament dynamics in cell motility Ifeaka Hangen-Mordi |
| title_short | The role of actin filament dynamics in cell motility |
| title_sort | the role of actin filament dynamics in cell motility |
| topic | Motilität (DE-588)4114638-4 gnd Dynamik (DE-588)4013384-9 gnd Actin-Filament (DE-588)4211337-4 gnd Zelle (DE-588)4067537-3 gnd |
| topic_facet | Motilität Dynamik Actin-Filament Zelle Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008768197&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT hangenmordiifeaka theroleofactinfilamentdynamicsincellmotility |