The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis:
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2012
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| Subjects: | |
| Online Access: | Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120206-1092917-1-1 Inhaltsverzeichnis |
| Physical Description: | X, 109 Bl. Ill., graph. Darst. |
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| 100 | 1 | |a Zhang, Lin |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis |c Lin Zhang |
| 264 | 1 | |c 2012 | |
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| adam_text | IMAGE 1
TABLE OF CONTENTS
PAGE NUMBER
CHAPTER 1: OVERVIEW
1.1 INTRODUCTION 1
1.2 OBJECTIVES AND SPECIFIC AIMS 1
1.3 REFERENCES 3
CHAPTER 2: BACKGROUND
2.1 S1P 5
2.1.1 S1P METABOLISM 5
2.1.2 S1P RECEPTORS 6
2.1.3 S1P GRADIENT 9
2.1.4 SPHINGOSINE KINASE 11
2.2 THROMBOPOIESIS 10
2.2.1 MEGAKARYOCYTE DIFFERENTIATION 11
2.2.2 PROPLATELET FORMATION 14
2.3 CONCLUSION 15
2.4 REFERENCES 16
CHAPTER 3: THE FUNCTION OF S1P RECEPTORS IN THROMBOPOIESIS
3.1 INTRODUCTION 24
3.2 MATERIAL AND METHODS 24
3.2.1 MATERIAL 24
3.2.2 MICE 24
3.2.3 CHIMERAS 25
3.2.4 LENTIVIRAL AND RETROVIRAL INFECTION 26
3.2.5 QUANTITATIVE RT-PCR 26
3.2.6 PLATELET MEASUREMENT 27
3.2.7 SERUM TPO MEASUREMENT 27
3.2.8 IMMUNOSTAINING 27
3.2.9 MEGAKARYOCYTE CULTURE 28
III
HTTP://D-NB.INFO/1031491848
IMAGE 2
3.2.10 MEGAKARYOCYTE COLONY-FORMING UNIT (CFU-MK) ASSAY 29
3.2.11 FLOW CYTOMETRY 29
3.2.12 FLUORESCENCE-ACTIVATED CELL SORTING (FACS) 30
3.2.13 SUBCLONING AND TRANSFECTION 30
3.2.14 PPF DIRECTION ASSAY 31
3. 2.15 WESTERN BLOT ANALYSIS 31
3.2.16 TWO-PHOTON INTRAVITAL IMAGING OF THE BONE MARROW 31
3.2.17 SHEAR STRESS 33
3.2.18 LIVE CELL IMAGING 34
3.2. 19 STATISTICAL ANALYSIS 34
3.3 RESULTS 34
3.3.1 S1P RECEPTOR EXPRESSION PROFILE IN MEGAKARYOCYTES 34
3.3.2 PLATELET COUNTS IN PERIPHERAL BLOOD 38
3.3.3 CONDITIONAL DELETION OF S1P1 IN MEGAKARYOCYTES 41
3.3.4 GAIN OF FUNCTION OF S1P1 IN S1 P1~ / MEGAKARYOCYTES 42
3.3.5 THE REDUCED PLATELETS CAUSED BY LOSS OF S1P1 INDEPENDENT OF MOUSE
BACKGROUND 43
3.3.6 MK MATURATION 44
3.3.7 MOTIBILITY AND DISTRIBUTION OF MEGAKARYOCYTES IN VIVO 46
3.3.8 PLATELET LIFE SPAN 49
3.3.9 TPO LEVELS AND TPO RECEPTOR EXPRESSION 50
3.3.10 PROPLATELET FORMATION IN VITRO 52
3.3.11 DEMARCATION MEMBRANE SYSTEM (DMS) 53
3.3.12 POLARIZATION OF PROPLATELET FORMATION 54
3.3.13 FORMATION OF SINUSOIDAL PROPLATELET IN VIVO 55
3.3.14 PROPLATELET FRAGMENTATION IN VITRO 58
3.3.15 PROPLATELET FRAGMENTATION IN VIVO 64
3.3.16 PHARMACOLOGICAL INHIBITION OF S1P1 IN VIVO 67
3.3.17 MOLECULAR MECHANISM 70
3.3.18 IN VIVO EFFECT OF S1P ANALOGUES 72
IV
IMAGE 3
3.4 DISCUSSION 76
3.5 REFERENCES 78
CHAPTER 4 EFFECT OF SPHINGOSINE KINASES ON THROMBOPOIESIS
4.1 INTRODUCTION 81
4.2 MATERIAL AND METHOD 81
4.2.1 MATERIAL 81
4.2.2 MICE 81
4.2.3 CHIMAERAS 81
4.2.4 WESTERN BLOT ANALYSIS 82
4.2.5 ADDTIIONAL METHODS 82
4.3 RESULTS 82
4.3.1 SPHINGOSINE KINASE EXPRESSION PROFILE IN MEGAKARYOCYTES 82
4.3.2 PLATELET COUNTS IN PERIPHERAL BLOOD 84
4.3.3 SEARCHING THE REASONS FOR THROMBOCYTOPENIA IN SPHK2^ MICE 85
4.3.4 EXTRAMEDULLAR THROMBOPOIESIS IN SPHK2 _/ MICE 87
4.3.5 POLYPLOIDIZATION OF MEGAKARYOCYTES 89
4.3.6 PROPLATELET FORMATION IN VITRO AND IN VIVO 90
4.3.7 PROPLATELET FRAGMENTATION IN VITRO 91
4.3.8 PROPLATELET SHEDDING IN VIVO 93
4.3.9 MOLECULAR MECHANISM 96
4.4 DISCUSSION 100
4.5 REFERENCES 102
APPENDIX
ABBREVIATIONS 104
LIST OF PRIMERS 107
LIST OF PUBLICATION 108
ACKNOWLEDG MENT
V
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| spelling | Zhang, Lin Verfasser aut The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis Lin Zhang 2012 X, 109 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier München, Techn. Univ., Diss., 2012 (DE-588)4113937-9 Hochschulschrift gnd-content Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20120206-1092917-1-1 http://mediatum.ub.tum.de/node?id=1092917 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120206-1092917-1-1 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024884027&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zhang, Lin The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis |
| subject_GND | (DE-588)4113937-9 |
| title | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis |
| title_auth | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis |
| title_exact_search | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis |
| title_full | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis Lin Zhang |
| title_fullStr | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis Lin Zhang |
| title_full_unstemmed | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis Lin Zhang |
| title_short | The effect of sphingosine 1-phosphate (S1P) on thrombopoiesis |
| title_sort | the effect of sphingosine 1 phosphate s1p on thrombopoiesis |
| topic_facet | Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1092917 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120206-1092917-1-1 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=024884027&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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