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Die Wirkung estrogen aktiver Umweltchemikalien auf Fische : Untersuchung reproduktionsrelevanter Parameter bei Regenbogenforelle und Zebrabärbling

Bieberstein, Ute

English Title: Effects of estrogenic chemicals on fish : investigation of reproduction parameters on rainbow trout and zebrafish

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Abstract

Ziel der Dissertation war es, eine In situ-Methode zum Nachweis von Vitellogenin (Vg) nach 17b-Estrogen- und Xenoestrogen-Induktion in der Leber männlicher Fische zu etablieren sowie die Wirkung verschiedener umweltrelevanter, estrogener Chemikalien auf eine Fischpopulation zu untersuchen. Hierzu wurden immunhisto- und immuncytochemische Untersuchungen an Regenbogenforellen (Oncorhynchus mykiss) und In vivo-Langzeitversuche mit Zebrabärblingen (Danio rerio) über den gesamten Lebenszyklus im Durchflusssystem durchgeführt. Mit Hilfe eines polyklonalen Antikörpers gegen Regenbogenforellen-Vg gelang es, nach 17b-Estradiol-Injektion Vg in der Leber weiblicher und männlicher Regenbogenforellen nachzuweisen. Durch die Immuncytochemie konnte der intrazelluläre Synthese- und Sekretionsweg von Vg über das rER und den Golgi-Apparat dargestellt werden. Am Beispiel des Zebrabärblings war die endokrine Wirkung von Nonylphenol (NP), Octylphenol (OP) und Dehydroepiandrosteron (DHEA) auf Fische nachweisbar. Die Untersuchung ausgewählter entwicklungs-biologischer, reproduktionsbiologischer, anatomischer, histologischer und ultrastruktureller Parameter ermöglichte eine Bewertung schadstoffbedingter Veränderungen während der Larval-, Juvenil- und Adultphase der Tiere. Die lebenslange Belastung der Zebrabärblings-Populationen mit NP, OP und DHEA führte grundsätzlich zu einer deutlichen Verschiebung des Geschlechterverhältnisses in Richtung der weiblichen Fische. Nach Abschluss der Exposition der Zebrabärblinge im Durchflusssystem waren, induziert durch NP, OP und DHEA, weitreichende konditionelle und strukturelle Veränderungen der Reproduktionsorgane, des Skeletts und der Leber dokumentierbar. Es konnte nachgewiesen werden, dass toxische Wirkungen von NP, OP und DHEA bereits nach einwöchiger Belastung während der Embryonal- und Larvalphase erkannt werden können, während sich estrogene Wirkungen auf den Zebrabärbling, erst nach Exposition über den gesamten Lebenszyklus zeigen.

Translation of abstract (English)

The aim of the following dissertation was firstly the establishment of a new in situ method to localize vitellogenin (Vg) in hepatocytes of male fish after external stimulation with 17b-estradiol and secondly to elucidate the in vivo potential for endocrine disruption of different chemicals on fish populations. To accomplish these goals , immunhisto- and immuncytochemical tests on rainbow trout (Oncorhynchus mykiss) and life-cycle tests were carried out with zebrafish (Danio rerio) in a flow-trough system. Vg could be visualized with a polyclonal antibody against rainbow trout Vg after 17b-estradiol injection in liver of male and female rainbow trout. On a light microscopical level , antibodies primarily bound to regions around the nucleus, which contain, among other organelles, rER and Golgi fields, where Vg is synthesized. On an ultrastructural level, these organelles could be confirmed as the sites of vitellogenin synthesis. An endocrine action of nonylphenol (NP), octylphenol (OP) and dehydroepiandrosterone (DHEA) could be demonstrated in zebrafish. Chemically induced developmental, reproductive, anatomical, histological and ultrastructural changes were detectable in larvae as well as juvenil and adult fish. Exposition of zebrafish populations to NP, OP and DHEA led to a shift in sex ratio towards females in all cases. Furthermore treatment with NP, OP and DHEA resulted in conditional and structural changes of reproductive organs, skeleton and liver of zebrafish. A toxic action of NP, OP and DHEA could be proven after one week of exposure during embryonal and larval phase, while an estrogenic action of the chemicals could not be demonstrated until the completion of the full life-cycle exposure.

Item Type: Dissertation
Supervisor: Dr. Braunbeck, Privatdoz Thomas
Date of thesis defense: 2. February 2001
Date Deposited: 20. Mar 2001 00:00
Date: 2000
Faculties / Institutes: Service facilities > Centre for Organismal Studies Heidelberg (COS)
Subjects: 570 Life sciences
Controlled Keywords: Fische, Xenobiotikum, Reproduktionstoxikologie, Hormon, Histopathologie
Uncontrolled Keywords: Teratogenität , Xenoestrogene , Immunhistochemie , Elektronenmikroskopie , Skelettefish, xenoestrogens, histopathology, reproduction, immunohistochemistry
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