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Prof. Dr. Med. Th. Zahnert Die Olfaktorische Wahrnehmung Von Bour

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Prof. Dr. Med. Th. Zahnert Die Olfaktorische Wahrnehmung Von Bour Aus der Klinik für Hals-, Nasen und Ohrenheilkunde Direktor: Prof. Dr. med. Th. Zahnert Die olfaktorische Wahrnehmung von Bourgeonal durch infertile und fertile Männer und Genotypisierung des Bourgeonalrezeptors hOR 17-4 Dissertationsschrift zur Erlangung eines doctor medicinae (Dr. med.) der Medizinischen Fakultät Carl Gustav Carus der Technischen Universität Dresden vorgelegt von Eva Kemper aus Würselen Berlin 2014 1. Gutachter: 2. Gutachter: Tag der mündlichen Prüfung: gez: ------------------------------------------------- Vorsitzender der Promotionskommission I Inhaltsverzeichnis 1. Einleitung................................................................................................................................. 1 1.1. Hintergrund ...................................................................................................................... 1 1.2. Problemstellung ............................................................................................................... 2 1.3. Grundlagen ...................................................................................................................... 2 1.3.1. Der Geruchssinn ........................................................................................................ 2 1.3.2. Das menschliche Riechrezeptorrepertoire ............................................................... 4 1.3.3. Ektopische Expression olfaktorischer Rezeptoren und Spermien-chemotaxis ....... 5 1.3.4. Der olfaktorische Rezeptor hOR 17-4 ....................................................................... 6 1.3.5. Bourgeonal als Riechrezeptorligand menschlicher Spermien .................................. 9 1.3.6. Rezeptorfunktion von hOR17-4 in menschlichen Spermatozoen .......................... 10 1.3.7. Olfaktologie .............................................................................................................. 12 1.3.8. Andrologie ............................................................................................................... 17 1.3.9. Variabilität des Genoms ......................................................................................... 20 1.4. Fragestellung .................................................................................................................. 21 2. Material und Methoden........................................................................................................ 22 2.1. Ethischer Hintergrund................................................................................................... 22 2.2. Probanden und Patienten ............................................................................................. 22 2.3. Studiendesign und Versuchsablauf ............................................................................... 23 2.4. Anamnese ...................................................................................................................... 24 2.5. Sniffin' Sticks ................................................................................................................. 25 2.5.1. Identifikation .......................................................................................................... 26 2.6. Diskriminations- und Schwellentestung ....................................................................... 28 2.6.1. Geruchsschwelle ..................................................................................................... 29 2.7. Subjektive Intensitätseinstufung ................................................................................... 30 2.8. Charakterisierung von Varianten des OR17-4 Gens ...................................................... 31 2.8.1. Experimenteller Ansatz ........................................................................................... 31 2.8.2. Agarosegelelektrophorese zur Auftrennung von DNA-Fragmenten ..................... 32 2.8.3. Präparation von DNA-Fragmenten aus Agarosegelen ........................................... 33 2.8.4. DNA- Sequenzierung ............................................................................................. 33 2.8.5. Isolierung genomischer DNA aus Blut nach dem 'QIAamp DNA Mini Kit Tissue Protocol' (Qiagen) ............................................................................................................ 34 2.8.6. Polymerase-Kettenreaktion ................................................................................... 34 2.9. Statistische Analysemethoden ...................................................................................... 36 3. Ergebnisse ............................................................................................................................ 37 II 3.1. Deskriptive Statistik....................................................................................................... 37 3.2. Screeningtest der Sniffin' Sticks .................................................................................... 37 3.3. Schwellen- und Diskriminationstestung ....................................................................... 38 3.3.1. Phenylethylalkohol ................................................................................................. 39 3.3.2. Bourgeonal ............................................................................................................. 39 3.3.3. Helional .................................................................................................................. 40 3.4. Subjektive Intensitätsschätzung ....................................................................................41 3.4.1. Phenylethylalkohol ..................................................................................................41 3.4.2. Bourgeonal ..............................................................................................................41 3.4.3. Helional .................................................................................................................. 42 3.5. Ergebnisse der Genotypisierung ................................................................................... 43 3.6. Zusammenfassung der Ergebnisse................................................................................ 43 3.6.1. Sniffin' Sticks .......................................................................................................... 43 3.6.2. Schwellen- und Diskriminationstestung ................................................................ 44 3.6.3. Intensitätsschätzung .............................................................................................. 44 3.6.4. Genotypisierung ..................................................................................................... 45 4. Diskussion ............................................................................................................................ 47 4.1. Riechtestungen .............................................................................................................. 47 4.2. Genotypisierung des Riechrezeptors hOR 17-4 ............................................................ 50 4.3. Die Bedeutung von hOR 17-4 für Fertilität und Reproduktionsmedizin ...................... 52 5. Zusammenfassung ................................................................................................................ 57 Summary ...................................................................................................................... 59 Erklärungen ...................................................................................................................61 6. Thesen .................................................................................................................................. 64 Literaturverzeichnis ..................................................................................................... 65 Abbildungsverzeichnis ................................................................................................. 82 Tabellenverzeichnis ...................................................................................................... 83 Anhang ......................................................................................................................... 84 Dank ............................................................................................................................. 90 III Abkürzungsverzeichnis Abb. Abbildung Aq. dest. Aqua destillata (Lat.: destilliertes Wasser) cAMP Cyclisches Adenosinmonophosphat CCCRC Connecticut Chemosensory Clinical Research Center CC-SIT Cross- Cultural Smell Indentification Test CSSEP Chemo-somatosensorisch evozierte Potentiale dNTP Desoxyribonukleosidtriphosphat EEG Elektroenzephalogramm Engl. Englisch Et al. et alii (Lat.: und andere) FSH Follikelstimulierendes Hormon GnRH Gonadotropin Releasing-Hormon GPCR G protein-coupled receptor (Engl. : G Protein-gekoppelter Rezeptor) hCG Humanes Choriongonadotropin HCl Chlorwasserstoff hMG Humanes Menopausengonadotropin/ Menotropin HNO Hals-Nasen-Ohrenheilkunde kDa Kilodalton Lat. Latein MHC Major histocompatibility complex (Engl.: Haupthistokompatibilitäts- komplex) min. Minute mg Milligramm ml Milliliter ms Millisekunden MRT Magnetresonanztomogramm µg Mikrogramm µl Mikroliter n Numerus (Lat.: Anzahl) N.; Nn. Nervus; nervi (Lat.: Nerv; Nerven) ng Nanogramm OD Optische Dichte OE Olfaktorisches Epithel OEP Olfaktorisch evozierte Potentiale OR/ ORN Olfaktorische Rezeptoren/ Olfaktorische Rezeptorneurone IV PCR Polymerase chain reaction
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