Thickness, dynamics and mechanics of the Actomyosin Cortex:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XVIII, 217 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Thickness, dynamics and mechanics of the Actomyosin Cortex |c von Andrew G. Clark |
| 264 | 1 | |c 2013 | |
| 300 | |a XVIII, 217 S. |b Ill., graph. Darst. | ||
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| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Dresden, Techn. Univ., Diss., 2013 | ||
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Datensatz im Suchindex
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| adam_text | CONTENTS
LIST OF FIGURES XV
LIST OF TABLES XVI
1 INTRODUCTION 1
1.1 CELL BIOPHYSICS 2
1.1.1 PHYSICAL PROPERTIES OF THE CELL 2
CORTICAL TENSION AND OSMOTIC PRESSURE 2
VISCOELASTIC PROPERTIES 5
MEMBRANE-TO-CORTEX ATTACHMENT 5
CELL ADHESION 6
1.1.2 PHYSICAL MEASUREMENTS IN LIVING CELLS 7
CORTICAL TENSION MEASUREMENTS 8
MEASUREMENT OF CELULAR VISCOELASTIC PROPERTIES 11
MEMBRANE-TO-CORTEX ATTACHMENT AND ADHESION MEASUREMENTS 13
1.2 THE ACTIN CYTOSKELETON 15
1.2.1 ACTIN 15
BIOCHEMICAL PROPERTIES 16
BIOPHYSICAL PROPERTIES 20
1.2.2 MYOSINS 23
SINGLE-MOLECULE PROPERTIES OF MYOSINS 24
CONVENTIONAL MYOSINS 27
UNCONVENTIONAL MYOSINS 29
1.2.3 ACTIN BINDING PROTEINS 30
ACTIN NUCLEATORS 31
ACTIN CAPPING PROTEINS 32
ACTIN SEVERING PROTEINS 34
G-ACTIN BINDING PROTEINS 35
IX
HTTP://D-NB.INFO/1042777772
ACTIN SIDEBINDING PROTEINS 36
ACTIN CROSS-LINKERS AND BUNDLERS 37
ACTIN-MEMBRANE LINKERS 38
1.2.4 UPSTREAM REGULATION OF ACTOMYOSIN 39
1.2.5 ACTOMYOSIN ASSEMBLIES 42
LAMELLIPODIA 43
THE ATOMYOSIN CORTEX 43
1.3 CORTEX-DRIVEN CELL SHAPE CHANGE 47
1.3.1 THE CELL CYCLE 47
SHAPE OSCILLATIONS IN EARLY AMPHIBIAN EMBRYOS 47
CELL ROUNDING 49
CYTOKINESIS 50
1.3.2 CELL MIGRATION 53
1.4 SUB-RESOLUTION IMAGE ANALYSIS 56
1.4.1 SINGLE MOLECULE HIGH-RESOLUTION COLOCALIZATION (SHREC) 56
1.4.2 MODEL-CONVOLUTION 58
1.5 AIMS 62
2 RESULTS I 63
2.1 CORTEX THICKNESS 64
2.1.1 RELATIVE LOCALIZATION OF THE ACTIN CORTEX AND PLASMA MEMBRANE 64
2.1.2 INTRACELLULAR ACTIN FLUORESCENCE INFLUENCES CORTEX-PLASMA MEMBRANE
PEAK
SEPARATION 67
2.1.3 EXTRACTION OF CORTEX THICKNESS USING A SIMPLE DESCRIPTION OF
CORTEX GEOMETRY 71
2.1.4 EXTRACTION OF CORTEX THICKNESS FROM CORTEX AND PLASMA MEMBRANE
LINESCANS 74
2.1.5 EVALUATION OF CHROMATIC SHIFT CORRECTION 78
2.1.6 EVALUATING CORTEX THICKNESS MEASUREMENTS BY MODEL-CONVOLUTION 82
2.1.7 ESTIMATING THE INFLUENCE OF MICROVILLI ON CORTEX THICKNESS
MEASUREMENTS . 85
2.1.8 CORTEX THICKNESS INCREASES DURING BLEB RETRACTION 90
2.1.9 MOLECULAR REGULATION OF CORTEX THICKNESS 92
ACTIN DISASSEMBLY AND CORTEX THICKNESS- 92
ACTIN NUCLEATOR ACTIVITY AND CORTEX THICKNESS 95
CORTEX THICKNESS DURING CELL CYCLE PROGRESSION AND THE EFFECT OF
MYOSIN-2
ACTIVITY 96
2.2 CORTEX DYNAMICS 99
2.2.1 CORTICAL ACTIN TURNOVER I: RECOVERY DYNAMICS AND ANALYSIS 100
OBSERVATION OF CORTICAL ACTIN DYNAMICS IN DIFFERENT CELL LINES 100
ANALYSIS OF CORTICAL ACTIN FRAP DATA 103
2.2.2 CORTICAL ACTIN TURNOVER II: REGULATION THROUGH THE CELL CYCLE AND
BY MYOSIN-2
ACTIVITY 108
CONTROL OF CORTICAL ACTIN TURNOVER DURING CELL CYCLE PROGRESSION 108
THE ROLE OF MYOSIN-2 ACTIVITY IN CORTICAL ACTIN TURNOVER 110
2.2.3 CORTICAL MYOSIN-2 TURNOVER 113
2.3 CORTEX MECHANICS 115
2.3.1 CORTICAL TENSION MEASUREMENTS IN HELA CELLS BY MICROPIPETTE
ASPIRATION . 115
2.3.2 MYOSIN-2 ACTIVITY AND CORTEX TENSION 122
3 DISCUSSION 125
3.1 CORTEX THICKNESS MEASUREMENTS 125
3.1.1 TECHNICAL DISCUSSION OF ASSUMPTIONS AND CAVEATS 127
THE INFLUENCE OF CORTICAL ACTIN DISTRIBUTION 127
EFFECTIVE MEMBRANE THICKNESS 128
EXTRACTION OF I
OU
T 128
RED/GREEN MAGNIFICATION DIFFERENCE CORRECTION 129
3.1.2 FUTURE DIRECTIONS AND OUTLOOK 130
3.2 CORTICAL ACTIN FRAP DYNAMICS 132
3.3 CORTEX THICKNESS, TURNOVER AND TENSION 133
3.3.1 CORTEX THICKNESS AND ACTIN TURNOVER 133
3.3.2 CORTEX THICKNESS AND TENSION 134
3.3.3 CORTICAL TENSION, ACTIN TURNOVER AND MYOSIN-2 ACTIVITY 135
3.4 CONCLUDING REMARKS 136
4 MATERIALS AND METHODS 138
4.1 CELL CULTURE AND IMAGING PREPARATION 138
4.1.1 CELL LINES 138
4.1.2 PLASMID TRANSFECTION 138
4.1.3 BLOCKING, DETACHMENT AND IMAGING 139
4.2 IMAGE PROCESSING 140
4.3 PLASMIDS AND SIRNA 141
4.3.1 DNA CONTRUCTS USED IN THIS STUDY 141
4.3.2 RNAI CONSTRUCTS AND TRANSFECTION 141
4.4 CHEMICAL INHIBITORS AND OTHER REAGENTS 142
4.5 WESTERN BLOTS 142
4.6 IMAGE AND DATA ANALYSIS: CORTEX THICKNESS MEASUREMENTS 143
4.6.1 SEGMENTATION AND LINESCAN EXTRACTION 143
4.6.2 EXTRACTION OF LINESCAN INTENSITY PARAMETERS 145
4.6.3 RED/GREEN MAGNIFICATION SHIFT 145
4.6.4 CORTEX THICKNESS EXTRACTION 148
4.7 SYNTHETIC IMAGE GENERATION AND ANALYSIS 149
4.8 IMAGE AND DATA ANALYSIS: FRAP 149
4.8.1 EXTRACTION OF INTENSITY DATA AND NORMALIZATION 149
4.8.2 FULL AND 1/2 RECOVERY TIME EXTRACTION 150
4.9 MICROPIPETTE ASPIRATION 151
4.9.1 PREPARATION OF CELLS FOR MPA 151
4.9.2 TENSION MEASUREMENTS 151
4.9.3 DYNAMIC TIMELAPSE EXPERIMENTS 152
4.10 STATISTICAL ANALYSES AND PLOTTING 152
REFERENCES 153
A SYNTHETIC IMAGES GENERATION 174
A.L CELL SEGMENTATION AND MEMBRANE PATHS 174
A.2 SYNTHETIC CORTICAL ACTIN 176
A.3 SYNTHETIC MICROVILLI . 177
A.4 PROCESSING AND DOWNSAMPLING 177
B PUBLISHED MANUSCRIPT:
MONITORING ACTIN CORTEX THICKNESS IN LIVE CELLS 178
C SUPPORTING METHODS FOR PUBLISHED MANUSCRIPT:
MONITORING ACTIN CORTEX THICKNESS IN LIVE CELLS 190
DECLARATION 217
|
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| spelling | Clark, Andrew G. 1985- Verfasser (DE-588)1038811317 aut Thickness, dynamics and mechanics of the Actomyosin Cortex von Andrew G. Clark 2013 XVIII, 217 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Dresden, Techn. Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026909438&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
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| title_auth | Thickness, dynamics and mechanics of the Actomyosin Cortex |
| title_exact_search | Thickness, dynamics and mechanics of the Actomyosin Cortex |
| title_full | Thickness, dynamics and mechanics of the Actomyosin Cortex von Andrew G. Clark |
| title_fullStr | Thickness, dynamics and mechanics of the Actomyosin Cortex von Andrew G. Clark |
| title_full_unstemmed | Thickness, dynamics and mechanics of the Actomyosin Cortex von Andrew G. Clark |
| title_short | Thickness, dynamics and mechanics of the Actomyosin Cortex |
| title_sort | thickness dynamics and mechanics of the actomyosin cortex |
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