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Der Recessus Olfactorius Von Bombina Orientalis: Eine Vergleichende Analyse

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Der Recessus Olfactorius Von Bombina Orientalis: Eine Vergleichende Analyse Der Recessus olfactorius von Bombina orientalis: Eine vergleichende Analyse Dissertation zur Erlangung des akademischen Grades einer Doktorin der Naturwissenschaften (Dr. rer. nat.) Fachbereich 10, Mathematik und Naturwissenschaften Universität Kassel vorgelegt von Sabrina Jordan Kassel, im August 2017 Tag der Disputation: 13.12.2017 Gutachter/in: Dr. Christine Nowack Prof. Dr. Rüdiger Wagner Weitere Prüfungskommissionsmitglieder: Prof. Dr. Monika Stengl Prof. Dr. Kurt Weising Prof. Dr. Ulrich Kutschera Für Eva. Danksagung Ich möchte mich an dieser Stelle in aller Herzlichkeit bei all jenen bedanken, die mich während der Entstehungszeit dieser Dissertation begleitet haben. Zuerst danke ich Dr. Christine Nowack für ihre außerordentliche Unterstützung, die sowohl auf fachlicher als auch auf persönlicher Ebene stets konstruktiv, wertschätzend und partnerschaftlich ausfiel. Bei Prof. Dr. Rüdiger Wagner bedanke ich mich ganz herzlich für die bereitwillige Übernahme des Zweitgutachtens. Ich bedanke mich außerdem bei den Leiter/-innen und Mitarbeiter/-innen folgender Fach- und Forschungsgebiete für den kollegialen Austausch, die Zusammenarbeit und insbesondere für zur Verfügung gestellte Materialien und Arbeitsplätze: Zellbiologie unter Prof. Dr. M. Maniak, Tier- physiologie unter Prof. Dr. M. Stengl, Pflanzenphysiologie unter Prof. Dr. U. Kutschera, Biochemie unter Prof. Dr. F. Herberg und Didaktik der Biologie unter Prof. Dr. J. Mayer. Außerdem danke ich den Professor/-innen und ehemaligen Fachgebietsleiter/-innen, die bereits ihren wohlverdienten Ruhestand angetreten haben: Prof. Dr. M. Schäfer, Prof. Dr. H. Zöltzer und Prof. Dr. W. Nellen. Zudem danke ich der Universität Kassel für die Gewährung eines dreijährigen Promotionsstipendiums. Bei meinen Kolleginnen und Freundinnen Dr. Carolin Wittmer und Katrin Schröder möchte ich mich für die zahllosen Stunden bedanken, während derer wir miteinander gearbeitet, gelacht und nicht zuletzt literweise Kaffee, kiloweise Kuchen und das eine oder andere Eis verdrückt haben. Ihr habt die letzten Jahre um ein Vielfaches verschönert und maßgeblich zu meiner Ausdauer beigetragen. Meinem Partner Benjamin Rudolf danke ich für seine fortwährende Unterstützung, seine Liebe und Fürsorge insbesondere während der letzten, anstrengenden Monate. Meiner kleinen Tochter Eva danke ich für die nötige Ablenkung durch ihr Chaos, ihren Witz, ihre Kreativität und für ihre bedingungslose Liebe. Du bist mein Antrieb. Teile dieser Arbeit wurden veröffentlicht in: Nowack, C., Jordan, S. und C. Wittmer. 2013. The Recessus Olfactorius: A Cryptic Olfactory Organ of Anuran Amphibians. In: M. L. East und M. Dehnhard (Hg.): Chemical Signals in Vertebrates 12. Springer New York; S. 37–48. Vorträge: Jordan, S. und C. Nowack. 2012. Show me yours – I'll show you mine! Lectinhistochemmical comparison of the olfactory epithelia of Bombina orientalis (Discoglossidae) and Xenopus muelleri (Pipidae). Deutscher Herpetologentag und SEH European Congress of Herpetology. Luxemburg und Trier. Jordan, S. und C. Nowack. 2012. Comparative characterization of the Recessus olfactorius, a previously unregarded olfatory organ of anuran amphibians. Proteomics Day 2012/Kick-off Meeting Promotionskolleg „Funktionelle Analyse von Modifikationen an Makromolekülen“. Rothenburg a. d. Fulda. Jordan, S. und C. Nowack. 2012. The olfactory organs of the frogs Bombina orientalis (Discoglossidae) and X. borealis (Pipidae) compared by lectin histochemistry. 105th Annual Meeting of the German Zoological Society und dem Symposium Olfaction across species and systems. Konstanz. Jordan, S. und C. Nowack. 2013. The first steps: Presence and distribution of G protein α subunits in the three olfactory organs of the Clawed Frog Xenopus borealis. Functionomics Day 2013 (Promotionskolleg). Fuldatal. Jordan, S. und C. Nowack. 2013. Presence and distribution of G protein alpha subunits in the three olfactory organs of the toad Bombina orientalis. 10th International Congress of Vertebrate Morphology. Barcelona. Poster: Jordan, S. und C. Nowack. 2012. Lectin histochemistry on the three olfactory epithelia of the toad Bombina orientalis (Anura, Amphibia) reveals a distinct staining pattern for the recessus olfactorius. 12th Meeting on Chemical Signals in Vertebrates (CsiV XII). Berlin. I Inhaltsverzeichnis Abkürzungsverzeichnis..............................................................................................................IV Abbildungsverzeichnis...............................................................................................................VI Tabellenverzeichnis.................................................................................................................VIII 1 Zusammenfassung.....................................................................................................................1 2 Grundlagen.................................................................................................................................3 2.1 Allgemeine Biologie und Systematik der Anura...................................................................3 2.2 Die Geruchsorgane adulter Anura: Anatomie.......................................................................6 2.2.1 „Normaler Typ“ mit Recessus olfactorius ....................................................................6 2.2.2 „Pipider Typ“ mit middle chamber epithelium..............................................................8 2.3 Die Geruchsorgane adulter Anura: Bau der Sinnesepithelien...............................................9 2.3.1 Das Sinnesepithel des Hauptgeruchsorgans................................................................10 2.3.2 Das vomeronasale Sinnesepithel.................................................................................11 2.3.3 Der Recessus olfactorius.............................................................................................12 2.3.4 Das middle chamber epithelium (Pipidae)..................................................................13 2.4 Die nasalen Drüsen adulter Anura.......................................................................................13 2.5 Die Funktion der Geruchsorgane........................................................................................14 2.6 Die Detektion olfaktorischer Stimuli..................................................................................15 2.6.1 Perireceptor events......................................................................................................16 2.6.2 Olfaktorische Rezeptorproteine und -genfamilien......................................................17 2.6.3 Coexpression von G-Protein-α-Untereinheiten und Duftstoffrezeptoren...................20 2.6.4 Transduktion des Signals und Zielregion im Gehirn...................................................21 2.7 Lektinhistochemie...............................................................................................................23 2.8 Zielsetzung der vorliegenden Untersuchung.......................................................................24 3 Material und Methoden...........................................................................................................26 3.1 Tiermaterial.........................................................................................................................26 3.2 Paraffineinbettung und Anfertigung von Schnittserien.......................................................27 3.3 Histologische Übersichtsfärbungen....................................................................................27 3.4 Aralditeinbettung und Anfertigung von Semidünnschnitten...............................................28 II 3.4.1 Fixierung und Vorbereitung des Gewebes...................................................................28 3.4.2 Kontrastierung, Entwässerung und Einbettung in Kunststoff (Araldit)......................29 3.4.3 Anfertigung und Färbung von Semidünnschnitten......................................................29 3.5 Lektinhistochemie...............................................................................................................30 3.5.1 Gewebevorbereitung und Durchführung der Färbung................................................30 3.5.2 Auswertung und Dokumentation.................................................................................32 3.6 Immunhistochemie..............................................................................................................34 3.6.1 Antikörper-Funktionstest.............................................................................................34 3.6.2 Immunmarkierung mit Anti-Gαolf, -Gαo und -Gαi2..................................................35 4 Ergebnisse.................................................................................................................................38 4.1 Histologie der Geruchssinnesepithelien: Paraffinschnitte..................................................38 4.1.1 Bombina orientalis......................................................................................................38 4.1.2 Xenopus borealis.........................................................................................................40 4.2 Histologie der Geruchssinnesepithelien: Semidünnschnitte...............................................42 4.3 Lektinhistochemie...............................................................................................................46 4.3.1 Bombina orientalis......................................................................................................46
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