Study of local protein synthesis in growth cones of embryonic mouse motor neurons:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | Volltext Inhaltsverzeichnis |
| Beschreibung: | 107 S. Ill., graph. Darst. |
Internformat
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| 100 | 1 | |a Rathod, Reenaben Jagdishbhai |d 1979- |e Verfasser |0 (DE-588)1024503542 |4 aut | |
| 245 | 1 | 0 | |a Study of local protein synthesis in growth cones of embryonic mouse motor neurons |c von Rathod Reenaben Jagdishbhai |
| 246 | 1 | 3 | |a Analyse der lokalen Proteinsynthese in Wachstumskegeln von embryonalen Maus Motoneuronen |
| 264 | 1 | |c 2012 | |
| 300 | |a 107 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Würzburg, Univ., Diss., 2012 | ||
| 610 | 2 | 7 | |a Embryo |g Musikgruppe : München |0 (DE-588)10196171-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Maus |0 (DE-588)4169148-9 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Proteinsynthese |0 (DE-588)4261878-2 |2 gnd |9 rswk-swf |
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| 689 | 0 | 1 | |a Embryo |g Musikgruppe : München |0 (DE-588)10196171-6 |D b |
| 689 | 0 | 2 | |a Motoneuron |0 (DE-588)4135747-4 |D s |
| 689 | 0 | 3 | |a Proteinsynthese |0 (DE-588)4261878-2 |D s |
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| 912 | |a ebook | ||
| 999 | |a oai:aleph.bib-bvb.de:BVB01-025183393 | ||
Datensatz im Suchindex
| _version_ | 1804149359946760192 |
|---|---|
| adam_text | IMAGE 1
CONTENTS
1 SUMMARY 5
2 ZUSAMMENFASSUNG 6
3 INTRODUCTION 8
3.1 VISUALIZATION OF LOCAL TRANSLATION 10
3.2 REGULATION OF LOCAL TRANSLATION BY EXTERNAL CUES 11
3.3 SPINAL MUSCULAR ATROPHY 11
3.4 LAMININ ISOFORMS AS SIGNALLING CUES IN THE EXTRA CELLULAR MATRIX 14
3.5 EMBRYONIC STEM CELL DIFFERENTIATION INTO MOTOR NEURONS 15
3.6 OBJECTIVE OF THE PRESENT STUDY 17
4 MATERIAL AND METHODS 18
4.1 CHEMICALS AND ENZYMES 18
4.2 ANIMALS 18
4.3 MATERIALS 18
4.3.1 CELL LINES 18
4.3.2 BUFFERS, SOLUTIONS AND MEDIUMS 18
4.3.3 FOR PRIMARY MOUSE MOTOR NEURON CULTURE 19
4.3.4 ANTIBODIES AND LABELING DYES 20
4.3.5 PLASMIDS, VECTORS AND LENTIVIRUSES MADE DURING THE COURSE OF WORK.
21
4.3.6 PRIMERS FOR CLONING 23
4.3.7 PRIMERS FOR RT 23
4.3.8 PRIMERS FOR SMN GENOTYPING 24
4.3.9 LNA PROBES FOR INSITU HYBRIDIZATION 24
4.3.10 LADDERS 24
4.3.11 KITS 24
4.3.12 FOR PRODUCTION OF ACTIVE SONIC HEDGEHOG 26
4.3.13 FOR ES CELL DIFFERENTIATION INTO MOTOR NEURONS 26
4.3.14 MICROSCOPES AND INSTRUMENTS 27
4.3.15 SOFTWARE 27
2
HTTP://D-NB.INFO/1025110293
IMAGE 2
4.4 METHODS 28
4.4.1 PRIMARY MOTOR NEURON CULTURING 28
4.4.2 GENOTYPING OF MOUSE EMBRYOS 28
4.4.3 CLONING 29
4.4.4 ISOLATION OF RNA 32
4.4.5 CDNA SYNTHESIS FROM RNA(REVERSE TRANSCRIPTION). 32
4.4.6 QUANTIATIVE PCR 33
4.4.7 MICROINJECTION 35
4.4.8 IN VITRO TRANSCRIPTION AND FLUORESCENCE LABELING OF MRNA 35
4.4.9 CALCIUM PRECIPITATE METHOD FOR TRANSFECTION OF MOTOR NEURONS 36
4.4.10 FRAP ON NIKONTE2000 36
4.4.11 MAGNETOFECTION OF MOTOR NEURONS 37
4.4.12 PRODUCTION OF LENTIVIRUSES 38
4.4.13 LENTIVIRAL INFECTION OF MOTOR NEURONS 39
4.4.14 FRAP ON SP5 39
4.4.15 DATA ANALYSIS 39
4.4.16 IN SITU HYBRIDIZATION 40
4.4.17 IMMUNOCYTOCHEMISTRY 40
4.4.18 DIFFERENTIATION OF ES CELLS INTO MOTOR NEURONS 40
4.4.19 ENRICHMENT OF DIFFERENTIATED MOTOR NEURONS BY A P75 ANTIBODY
PANNING STEP 41
5 RESULTS 42
5.1 MICROINJECTION OF LABELED P-ACTIN MRNA TO VISUALIZE TRANSPORT OF
MRNPS. 42
FIGURE 4 42
5.2 LOCAL TRANSLATION OF P-ACTIN 43
5.3 OPTIMIZATION OF CONSTRUCTS FOR EXPRESSION OF THE 3 UTR OF THE
P-ACTIN TO EXAMINE
LOCAL TRANSLATION IN AXONAL GROWTH CONES OF MOTONEURONS 47
5.4 LOCAL PROTEIN SYNTHESIS IN AXONS OF MOUSE MOTONEURONS 57
5.5 LOCAL PROTEIN SYNTHESIS IN GROWTH CONES OF EMBRYONIC MOUSE
MOTONEURONS IS
REGULATED BY LAMININ SIGNALLING 59
5.6 DEREGULATION OF LOCAL PROTEIN SYNTHESIS IN SMN / ;SMN2 CELLS 60
3
IMAGE 3
5.7 STANDARDIZATION OF IN SITU AGAINST EGFP WITH LNA PROBES. 64
5.8 LOCALIZATION OF THE EGFP CONTAINING MRNA 67
5.9 LOCALIZATION OF MRNA IS AFFECTED BY SMN LEVELS BUT NOT BY LAMININ
SIGNALING 69
5.10 AXON LENGTH MEASUREMENT 71
5.11 ENDOGENOUS ACTIN ISOFORM EXPRESSION 72
5.11.1 IN S/MIYBRIDIZATIONFOR ANALYSIS OF SUBCELLULAR DISTRIBUTION OF
ENDOGENOUS ACTIN
ISOFORMS 74
5.12 CLONING OF A REPORTER CONSTRUCT FOR A-ACTIN 3 UTR 78
5.13 PRODUCTION OF ACTIVE SONIC HEDGEHOG 79
5.14 DIFFERENTIATION OF EMBRYONIC STEM CELLS INTO MOTONEURONS 81
6 DISCUSSION 86
6.1 OPTIMIZATION OF CONSTRUCT FOR STUDYING LOCAL TRANSLATION IN
MOTONEURONS 86
6.2 LOCAL TRANSLATION IN MOUSE MOTONEURONS 86
6.3 TRANSPORT OF (3-ACTIN MRNA TO AXON GROWTH CONES AND ROLE OF SMN 87
6.4 ANALYSIS OF LOCAL TRANSLATION IN RESPONSE TO LAMININ 1 AND LAMININ 2
AND ROLE OF
SMN IN MOTONEURONS 87
6.5 ANALYSIS OF LOCAL TRANSLATION OF REPORTER CONSTRUCTS WITH ZIP CODE
IN MOTOR CONTROL
AND SMA MOTONEURONS 88
6.6 TRANSPORT AND TRANSLATION OF OTHER ACTIN ISOFORMS IN SMA 88
6.7 REGULATION OF (3-ACTIN LOCAL TRANSLATION BY MRNA BINDING PROTEINS 90
6.8 SIGNALLING PATHWAYS FOR REGULATION OF LOCAL TRANSLATION 91
6.9 ES CELL DERIVED MOTONEURONS 93
7 ABBREVIATIONS 95
8 REFRENCES 97
9 CURRICULUM VITAE 103
10 PUBLICATIONS 105
11 ACKNOWLEDGEMENTS 106
4
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| indexdate | 2024-07-10T00:21:47Z |
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| language | English |
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| spelling | Rathod, Reenaben Jagdishbhai 1979- Verfasser (DE-588)1024503542 aut Study of local protein synthesis in growth cones of embryonic mouse motor neurons von Rathod Reenaben Jagdishbhai Analyse der lokalen Proteinsynthese in Wachstumskegeln von embryonalen Maus Motoneuronen 2012 107 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Würzburg, Univ., Diss., 2012 Embryo Musikgruppe : München (DE-588)10196171-6 gnd rswk-swf Maus (DE-588)4169148-9 gnd rswk-swf Proteinsynthese (DE-588)4261878-2 gnd rswk-swf Motoneuron (DE-588)4135747-4 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Maus (DE-588)4169148-9 s Embryo Musikgruppe : München (DE-588)10196171-6 b Motoneuron (DE-588)4135747-4 s Proteinsynthese (DE-588)4261878-2 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-72045 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-72045 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025183393&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Rathod, Reenaben Jagdishbhai 1979- Study of local protein synthesis in growth cones of embryonic mouse motor neurons Embryo Musikgruppe : München (DE-588)10196171-6 gnd Maus (DE-588)4169148-9 gnd Proteinsynthese (DE-588)4261878-2 gnd Motoneuron (DE-588)4135747-4 gnd |
| subject_GND | (DE-588)10196171-6 (DE-588)4169148-9 (DE-588)4261878-2 (DE-588)4135747-4 (DE-588)4113937-9 |
| title | Study of local protein synthesis in growth cones of embryonic mouse motor neurons |
| title_alt | Analyse der lokalen Proteinsynthese in Wachstumskegeln von embryonalen Maus Motoneuronen |
| title_auth | Study of local protein synthesis in growth cones of embryonic mouse motor neurons |
| title_exact_search | Study of local protein synthesis in growth cones of embryonic mouse motor neurons |
| title_full | Study of local protein synthesis in growth cones of embryonic mouse motor neurons von Rathod Reenaben Jagdishbhai |
| title_fullStr | Study of local protein synthesis in growth cones of embryonic mouse motor neurons von Rathod Reenaben Jagdishbhai |
| title_full_unstemmed | Study of local protein synthesis in growth cones of embryonic mouse motor neurons von Rathod Reenaben Jagdishbhai |
| title_short | Study of local protein synthesis in growth cones of embryonic mouse motor neurons |
| title_sort | study of local protein synthesis in growth cones of embryonic mouse motor neurons |
| topic | Embryo Musikgruppe : München (DE-588)10196171-6 gnd Maus (DE-588)4169148-9 gnd Proteinsynthese (DE-588)4261878-2 gnd Motoneuron (DE-588)4135747-4 gnd |
| topic_facet | Embryo Musikgruppe : München Maus Proteinsynthese Motoneuron Hochschulschrift |
| url | https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-72045 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025183393&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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