Intracardiac valve cell differentiation in Drosophila melanogaster: histological and molecular analysis
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2014
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | 143 S. Ill., graph. Darst. 21 cm |
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| adam_text | UNIVERSITY
OSNABROCK
INTRACARDIAC VALVE CELL DIFFERENTIATION IN
DROSOPHILA MELANOGASTER:
HISTOLOGICAL AND MOLECULAR ANALYSIS
DISSERTATION
ZUR ERLANGUNG DES AKADEMISCHEN GRADES
DOKTOR DER NATURWISSENSCHAFTEN
(DR. RER. NAT.)
FACHBEREICH BIOLOGIE/CHEMIE DER UNIVERSITAT OSNABRUCK
VORGELEGT VON
BETTINA ABELN
OSNABRIICK, APRIL 2014
HTTP://D-NB.INFO/1053684541
TABLE OF CONTENTS
TABLE OF CONTENTS
1.1 INTRODUCTION 1
1.1.1 VALVE CELL PROGENITORS DEVELOP FROM MESODERMAL GERM LAYER 3
1.1.2 THE DORSAL VESSEL IS ESTABLISHED BY A REPETITIVE PATTERN OF HEART
CELLS 4
1.1.3 INTRACARDIAC VALVES OF THE DROSOPHILA HEART 5
1.1.4 TWO
ADDITIONAL VALVES DEVELOP DURING METAMORPHOSIS 5
1.1.5 LARVAL AND ADULT VALVE CELLS SHARE THE SAME HISTOLOGICAL
PROPERTIES 7
1.1.6 GENE EXPRESSION AND SIGNALING PATHWAYS OF VALVE CELLS 7
1.1.7 THE ROLE
OF HOMEOTIC GENE EXPRESSION IN VALVE CELL DETERMINATION 8
1.1.8 MAINTENANCE OF UNIDIRECTIONAL BLOOD FLOW AS A POTENTIAL FUNCTION
OF VALVE CELLS.. 10
1.2 WORK OBJECTIVES 11
2. MATERIAL AND METHODS 13
2.1 MATERIAL 13
2.1.1 FLY STRAINS....: 13
2.1.2 PRIMARY ANTIBODIES 22
2.1.3 SECONDAIY ANTIBODIES 23
2.1.4 FLUORESCENT/NON FLUORESCENT DYES 24
2.1.5 SOFTWARE 24
2.2 METHODS 25
2.2.1 FLY KEEPING AND HANDLING 25
2.2.2 ESTABLISHMENT OF THE FLY LINE 7B//305-GAL4; TOLLIOI-GFP 28
2.2.3. IN VITRO TRANSCRIPTION - RNA PROBES 31
2.2.4. IMMUNOHISTOCHEMISTRY 32
2.2.5 IN SITU HYBRIDIZATION 33
2.2.6 CRYO-SECTIONING 35
2.2.7 TEM ANALYSIS 36
TABLE OF CONTENTS
3. RESULTS 37
3.1 A NEW MARICER FOR CARDIOVASCULAR
VALVE DEVELOPMENT 37
3.2 ESTABLISHING THE TRANSGENIC FLY LINE 7B/OOJ-GAL4; TOLL305-GFP 40
3.3 CARDIAC VALVE CELLS ARISE BY DIFFERENTIATION OF TWO CARDIOMYOCYTES
DURING EARLY
LARVAL STAGES 41
3.3.1 HISTOLOGICAL ANATOMY OF EMBRYONIC (STAGE 16/17) AND 1
ST
INSTAR LARVAL HEARTS 41
3.3.2 TEM ANALYSES OF 2
ND
AND 3
1
INSTAR LARVAE VALVE CELLS 43
3.4 THE ORIGIN OF VALVOSOMES
- AN ULTRASTRUCTURAL ANALYSIS 44
3.5 ADULT VALVOSOMES CONTAIN COMPONENTS OF EXTRACELLULAR MATRIX 49
3.6 CRYO
SECTIONING CONFIRMS THE VIKING EXPRESSION PATTERN IN LARVAL AND ADULT
VALVES 51
3.7 TOLL MRNA AS WELL AS THE TOLL PROTEIN ARE EXPRESSED IN THE
EMBRYONIC, BUT NOT IN THE
3
RD
INSTAR LARVAL HEART 52
3.8 THE
GENE TOLL IS NOT INVOLVED IN CARDIOVASCULAR VALVE DEVELOPMENT 54
3.9 OVEREXPRESSION OF THE HOMEOTIC GENE ABDOMINAL-A INDUCES EXTRA VALVES
IN THE 3
1
INSTAR LARVAL HEART 56
3.10 THE PDGF/VEGF RECEPTOR PVR AND THE LIGAND PVF2 ARE EXCLUSIVELY
EXPRESSED IN
VALVE CELLS OF 3
RI
INSTAR
LARVAE 58
3.11 AUTOCRINE PVR AND PVF2 SIGNALING IS CRUCIAL FOR LARVAL VALVE CELL
DIFFERENTIATION 62
3.12 SHAPING THE CELL - HEART SPECIFIC RNAI MEDIATED CANDIDATE KNOCK
DOWN SCREEN ...66
3.12.1 CLATHRIN DEPENDENT ENDOCYTOSIS IS CRUCIAL FOR PROPER VALVE CELL
DEVELOPMENT 67
3.12.2 VALVOSOME MATURATION IS DYNAMIN DEPENDENT 69
3.12.3 RAB 5 IS INVOLVED IN VALVOSOME INITIATION - RAB 7 AND RAB 11
FUNCTION IN
VALVOSOME MATURATION 70
3.12.4 CLATHRIN INDEPENDENT PATHWAYS SEEM TO BE INVOLVED IN VALVOSOME
MATURATION . 71
3.12.4.1 CAVEOLAE FORMATION COULD BE INVOLVED IN VALVOSOME MATURATION 72
3.12.4.2 MACROPINOSOMES MIGHT BE NECESSAIY FOR GROWTH OF VALVOSOMES 72
3.12.4.3 THE CLIC/GEEC PATHWAY IS NOT INVOLVED IN VALVE CELL FORMATION
72
3.12.5 KNOCK DOWN OF SAR I AND SEC 23, COMPONENTS OF THE COP II PATHWAY,
RESULTED IN
MALFORMED VALVE CELLS 74
TABLE OF CONTENTS
3.12.6 COP I COATED VESICLE TRAFFICKING IS NOT INVOLVED IN VALVOSOME
FORMATION 75
3.12.7 VALVOSOME GROWTH IS ACHIEVED BY VESICLE FUSION MEDIATED BY SNARES
76
3.12.8 COR
VET AND HOPS COMPLEXES ARE INVOLVED IN VALVOSOME MATURATION 78
3.13 THE VESICLE FUSION PHENOTYPE AND THE MALFORMATION PHENOTYPE OF
KNOCK DOWN
ANIMALS (RAB 5, SAR 1, SHIBIRE) WERE CONFIRMED BY ULTRASTRUCTURAL TEM
ANALYSIS ...80
4. DISCUSSION 82
4.1 PDGF/VEGF SIGNALING IS IMPORTANT FOR VALVE CELL DIFFERENTIATION IN
DROSOPHILA AND
VERTEBRATES 86
4.2 VALVE CELL DEVELOPMENT IS REGULATED BY THREE IMPORTANT STEPS 89
4.3 THE
CHARACTERISTIC MORPHOLOGY OF DROSOPHILA VALVE CELLS IS UNIQUE IN
NATURE 90
5. CONCLUSIONS 92
6. SUMMARY 95
6.1 ZUSAMMENFASSUNG 96
6.2 RESUMEN 97
7. REFERENCES 98
8. PUBLICATIONS ILL
9. SUPPLEMENTAL MATERIAL 128
10. CURRICULUM VITAE 139
11. STATUTORY DECLARATION 141
12. ACKNOWLEDGEMENTS 142
|
| any_adam_object | 1 |
| author | Abeln, Bettina 1983- |
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| author_facet | Abeln, Bettina 1983- |
| author_role | aut |
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| bvnumber | BV042042170 |
| ctrlnum | (OCoLC)887785632 (DE-599)DNB1053684541 |
| dewey-full | 573.1715774 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 573 - Specific physiological systems in animals |
| dewey-raw | 573.1715774 |
| dewey-search | 573.1715774 |
| dewey-sort | 3573.1715774 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| format | Thesis Book |
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| spelling | Abeln, Bettina 1983- Verfasser (DE-588)1053816022 aut Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis vorgelegt von Bettina Abeln 2014 143 S. Ill., graph. Darst. 21 cm txt rdacontent n rdamedia nc rdacarrier Osnabrück, Univ., Diss., 2014 (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=027483403&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Abeln, Bettina 1983- Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis |
| subject_GND | (DE-588)4113937-9 |
| title | Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis |
| title_auth | Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis |
| title_exact_search | Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis |
| title_full | Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis vorgelegt von Bettina Abeln |
| title_fullStr | Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis vorgelegt von Bettina Abeln |
| title_full_unstemmed | Intracardiac valve cell differentiation in Drosophila melanogaster histological and molecular analysis vorgelegt von Bettina Abeln |
| title_short | Intracardiac valve cell differentiation in Drosophila melanogaster |
| title_sort | intracardiac valve cell differentiation in drosophila melanogaster histological and molecular analysis |
| title_sub | histological and molecular analysis |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027483403&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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