On the localization and physiological function of bitter taste receptors in gastrointestinal tract:
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 89 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a On the localization and physiological function of bitter taste receptors in gastrointestinal tract |c von Marta Bromke |
| 264 | 1 | |c 2013 | |
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| 502 | |a Potsdam, Univ., Diss., 2013 | ||
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Datensatz im Suchindex
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| adam_text | IMAGE 1
TABLE OF CONTENT
LIST OF FIGURES 1
LIST OF ABBREVIATIONS 2
1. INTRODUCTION 4
1.1. THE SENSE OF TASTE 4
1.2. TASTE PERCEPTION IN THE ORAL CAVITY 5
1.3. TASTE SIGNALLING CASCADE 7
1.4. EXTRAORAL TASTE RECEPTORS 9
1.5. INTESTINAL CELL TYPES AND THE PUTATIVE TASTE-LIKE CELLS IN THE GUT
9
1.6. GASTROINTESTINAL TAS1R RECEPTORS 11
1.7. GASTROINTESTINAL TAS2R RECEPTORS 12
1.8. THE AIM OF THE THESIS 14
2. MATERIALS AND METHODS 15
2.1. MATERIALS 15
2.1.1. CHEMICALS 15
2.1.2. ENZYMES 15
2.1.3. KITS 16
2.1.4. BACTERIAL MEDIA 16
2.1.5. CELL CULTURE MEDIA 16
2.1.6. BACTERIA 17
2.1.7. EUKARYOTIC CELLS 17
2.1.8. MICE 17
2.1.9. PLASMIDS 17
2.1.10. OLIGONUCLEOTIDES 17
2.1.11. ANTIBODIES 22
2.1.12. EQUIPMENT 23
2.1.13. SOFTWARE AND INTERNET RESOURCES 23
2.2. METHODS 24
2.2.1. CELL CULTURE METHODS 24
2.2.1.1. NC1-H716 CELL LINE CULTURE CONDITIONS 24
2.2.1.2. HEK 293T GAI 6GUST44 CELL LINE CULTURE CONDITIONS 25
2.2.1.3. FREEZING, STORAGE AND THAWING OF CELL LINES 25
2.2.2. FUNCTIONAL ANALYSIS OF BITTER TASTE RECEPTORS IN NCI-H716 CELLS
25
2.2.2.1. GLP-1 SECRETION STUDIES 25
2.2.2.2. CA 2+IMAGING 26
2.2.3. IMMUNODETECTION OF HETEROLOGOUSLY EXPRESSED MOUSE BITTER TASTE
RECEPTOR TAS2RL38 IN HEK239 CELLS 28
2.2.3.1. TRANSIENT TRANSFECTION 28
2.2.3.2. IMMUNOCYTOCHEMISTRY 29
2.2.4. ANIMALS 30
2.2.4.1. ANIMAL CARE AND MAINTENANCE 30
2.2.4.2. TISSUE COLLECTION AND FIXATION 30
2.2.5. MOLECULAR BIOLOGY METHODS 31
2.2.5.1. DNAISOLATION 31
2.2.5.2. QUANTIFICATION OF NUCLEIC ACIDS BY SPECTROPHOTOMETRY 32
2.2.5.3. RNA ISOLATION FROM NCI-H716 CELLS 32
2.2.5.4. RNA ISOLATION FROM MOUSE TISSUE 33
2.2.5.5. PRECIPITATION OF RNA 33
2.2.5.6. POLY A+ RNA ISOLATION 33
2.2.5.7. CDNA SYNTHESIS 33
2.2.5.8. POLYMERASE CHAIN REACTION 34
HTTP://D-NB.INFO/1042842442
IMAGE 2
2.2.5.9. AGAROSE GEL ELECTROPHORESIS 35
2.2.5.10. DNA RESTRICTION ENZYME DIGESTION 35
2.2.5.11. DNA PURIFICATION FROM AGAROSE GEL 35
2.2.5.12. LIGATION 36
2.2.5.13. HEAT-SHOCK TRANSFORMATION OF E.COLI BACTERIA 36
2.2.5.14. E.COLI BACTERIA CULTURE AND PLASMID ISOLATION 36
2.2.5.15. DNA SEQUENCE ANALYSIS 36
2.2.5.16. IN VITRO TRANSCRIPTION 37
2.2.6. HISTOLOGY METHODS 38
2.2.6.1. SECTIONING 38
2.2.6.2. IMMUNOHISTOCHEMISTRY 38
2.2.6.3. CONFOCAL MICROSCOPY 39
2.2.6.4. IN SITU HYBRIDIZATION 39
3. RESULTS 41
3.1. INVESTIGATION OFTAS2R FUNCTIONALITY IN HUMAN GUT-DERIVED NCI-H716
CELL LINE 41
3.1.1. EXPRESSION OF BITTER TASTE RECEPTORS AND MOLECULES OF TASTE
SIGNALLING CASCADE 41
3.1.2. GLP-1 SECRETION IN RESPONSE TO BITTER SUBSTANCES 43
3.1.3. FUNCTIONAL ANALYSIS OF TAS2RS IN NC1-H716 CELLS BY CA 2+ IMAGING
44
3.2. THE ANALYSIS OFGASTROINTESTINAL TRACT OF WILD TYPE C5 7BI/6 MICE TO
IDENTIFY BITTER TASTE RECEPTOR EXPRESSING CELLS 51
3.2.1. EXPRESSION OF TAS2RS AND MAIN TASTE SIGNALLING PATHWAY COMPONENTS
IN MOUSE GI TISSUES 51 3.2.2. DETECTING BITTER TASTE RECEPTOR TAS2RL 38
BY IMMUNOHISTOCHEMISTRY 53
3.2.3. DETECTING BITTER TASTE RECEPTORS BY IN SITU HYBRIDIZATION 55
3.2.4. ANALYSIS OF TAS2R EXPRESSION IN BLOOD 56
3.3. CHARACTERIZATION OF GENE-TARGETED MICE WITH MODIFIED TAS2RL31 LOCUS
57
3.3.1. ANALYSIS OF HRGFP EXPRESSION IN THE GUT OF TAS2RL3L LRGF7 MICE
58
3.3.2. HRGFP FLUORESCENCE IN GI TISSUES OF TAS2RL31 /,RGFP MICE 59
3.3.3. HRGFP MRNA DETECTION BY IN SITU HYBRIDIZATION 59
3.3.4. ANALYSIS OF CRE RECOMBINASE EXPRESSION IN THE GUT OF TAS2RL3 L
CRE MICE 60
3.3.5. EXPRESSION OF CRE RECOMBINASE IN DISTAL GUT OF THE TAS2RL31 CRE
MICE 62
4. DISCUSSION 63
4.1. ARE BITTER RECEPTORS EXPRESSED ON TISSUE LEVEL? 63
4.2. ARE BITTER RECEPTORS DETECTABLE IN SPECIFIC CELL TYPES? 65
4.3. CAN PHYSIOLOGICAL FUNCTIONS BE CONFIRMED IN A TAS2R-EXPRESSING
MODEL CELL LINE? 68
4.4. A PUTATIVE PHYSIOLOGICAL ROLE OF THE INTESTINAL BITTER TASTE
RECEPTORS 70
4.5. SUMMARY AND OUTLOOK. 72
5. LITERATURE 74
6. APPENDIX 87
6.1. ACKNOWLEDGEMENTS 87
6.2. CURRICULUM VITAE 88
6.3. DECLARATION BLQD! NIE ZDEFINIOWANO ZAKLADKI.
|
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| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
| dewey-search | 610 |
| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| spelling | Bromke, Marta 1982- Verfasser (DE-588)1038036984 aut On the localization and physiological function of bitter taste receptors in gastrointestinal tract von Marta Bromke 2013 89 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Potsdam, 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=026183835&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Bromke, Marta 1982- On the localization and physiological function of bitter taste receptors in gastrointestinal tract |
| subject_GND | (DE-588)4113937-9 |
| title | On the localization and physiological function of bitter taste receptors in gastrointestinal tract |
| title_auth | On the localization and physiological function of bitter taste receptors in gastrointestinal tract |
| title_exact_search | On the localization and physiological function of bitter taste receptors in gastrointestinal tract |
| title_full | On the localization and physiological function of bitter taste receptors in gastrointestinal tract von Marta Bromke |
| title_fullStr | On the localization and physiological function of bitter taste receptors in gastrointestinal tract von Marta Bromke |
| title_full_unstemmed | On the localization and physiological function of bitter taste receptors in gastrointestinal tract von Marta Bromke |
| title_short | On the localization and physiological function of bitter taste receptors in gastrointestinal tract |
| title_sort | on the localization and physiological function of bitter taste receptors in gastrointestinal tract |
| topic_facet | Hochschulschrift |
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