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Article: Fluorescence in situ hybridization techniques (FISH) to detect changes in CYP19a gene expression of Japanese medaka (Oryzias latipes)

TitleFluorescence in situ hybridization techniques (FISH) to detect changes in CYP19a gene expression of Japanese medaka (Oryzias latipes)
Authors
Issue Date2008
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/taap
Citation
Toxicology And Applied Pharmacology, 2008, v. 232 n. 2, p. 226-235 How to Cite?
AbstractThe aim of this study was to develop a sensitive in situ hybridization methodology using fluorescence-labeled riboprobes (FISH) that allows for the evaluation of gene expression profiles simultaneously in multiple target tissues of whole fish sections of Japanese medaka (Oryzias latipes). To date FISH methods have been limited in their application due to autofluorescence of tissues, fixatives or other components of the hybridization procedure. An optimized FISH method, based on confocal fluorescence microscopy was developed to reduce the autofluorescence signal. Because of its tissue- and gender-specific expression and relevance in studies of endocrine disruption, gonadal aromatase (CYP19a) was used as a model gene. The in situ hybridization (ISH) system was validated in a test exposure with the aromatase inhibitor fadrozole. The optimized FISH method revealed tissue-specific expression of the CYP19a gene. Furthermore, the assay could differentiate the abundance of CYP19a mRNA among cell types. Expression of CYP19a was primarily associated with early stage oocytes, and expression gradually decreased with increasing maturation. No expression of CYP19a mRNA was observed in other tissues such as brain, liver, or testes. Fadrozole (100 μg/L) caused up-regulation of CYP19a expression, a trend that was confirmed by RT-PCR analysis on excised tissues. In a combination approach with gonad histology, it could be shown that the increase in CYP19a expression as measured by RT-PCR on a whole tissue basis was due to a combination of both increases in numbers of CYP19a-containing cells and an increase in the amount of CYP19a mRNA present in the cells. © 2008 Elsevier Inc. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/179084
ISSN
2015 Impact Factor: 3.847
2015 SCImago Journal Rankings: 1.593
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorPark, JWen_US
dc.contributor.authorTompsett, Aen_US
dc.contributor.authorZhang, Xen_US
dc.contributor.authorNewsted, JLen_US
dc.contributor.authorJones, PDen_US
dc.contributor.authorAu, Den_US
dc.contributor.authorKong, Ren_US
dc.contributor.authorWu, RSSen_US
dc.contributor.authorGiesy, JPen_US
dc.contributor.authorHecker, Men_US
dc.date.accessioned2012-12-19T09:51:51Z-
dc.date.available2012-12-19T09:51:51Z-
dc.date.issued2008en_US
dc.identifier.citationToxicology And Applied Pharmacology, 2008, v. 232 n. 2, p. 226-235en_US
dc.identifier.issn0041-008Xen_US
dc.identifier.urihttp://hdl.handle.net/10722/179084-
dc.description.abstractThe aim of this study was to develop a sensitive in situ hybridization methodology using fluorescence-labeled riboprobes (FISH) that allows for the evaluation of gene expression profiles simultaneously in multiple target tissues of whole fish sections of Japanese medaka (Oryzias latipes). To date FISH methods have been limited in their application due to autofluorescence of tissues, fixatives or other components of the hybridization procedure. An optimized FISH method, based on confocal fluorescence microscopy was developed to reduce the autofluorescence signal. Because of its tissue- and gender-specific expression and relevance in studies of endocrine disruption, gonadal aromatase (CYP19a) was used as a model gene. The in situ hybridization (ISH) system was validated in a test exposure with the aromatase inhibitor fadrozole. The optimized FISH method revealed tissue-specific expression of the CYP19a gene. Furthermore, the assay could differentiate the abundance of CYP19a mRNA among cell types. Expression of CYP19a was primarily associated with early stage oocytes, and expression gradually decreased with increasing maturation. No expression of CYP19a mRNA was observed in other tissues such as brain, liver, or testes. Fadrozole (100 μg/L) caused up-regulation of CYP19a expression, a trend that was confirmed by RT-PCR analysis on excised tissues. In a combination approach with gonad histology, it could be shown that the increase in CYP19a expression as measured by RT-PCR on a whole tissue basis was due to a combination of both increases in numbers of CYP19a-containing cells and an increase in the amount of CYP19a mRNA present in the cells. © 2008 Elsevier Inc. All rights reserved.en_US
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/taapen_US
dc.relation.ispartofToxicology and Applied Pharmacologyen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAromatase - Analysis - Biosynthesis - Geneticsen_US
dc.subject.meshGene Expression Profilingen_US
dc.subject.meshGene Expression Regulation, Enzymologic - Genetics - Physiologyen_US
dc.subject.meshIn Situ Hybridization, Fluorescence - Methodsen_US
dc.subject.meshModels, Geneticen_US
dc.subject.meshOryzias - Genetics - Metabolismen_US
dc.subject.meshZebrafish Proteins - Analysis - Biosynthesis - Geneticsen_US
dc.titleFluorescence in situ hybridization techniques (FISH) to detect changes in CYP19a gene expression of Japanese medaka (Oryzias latipes)en_US
dc.typeArticleen_US
dc.identifier.emailWu, RSS: rudolfwu@hku.hken_US
dc.identifier.authorityWu, RSS=rp01398en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.taap.2008.06.012en_US
dc.identifier.pmid18644401-
dc.identifier.scopuseid_2-s2.0-52049083659en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-52049083659&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume232en_US
dc.identifier.issue2en_US
dc.identifier.spage226en_US
dc.identifier.epage235en_US
dc.identifier.isiWOS:000260290000008-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridPark, JW=8414402800en_US
dc.identifier.scopusauthoridTompsett, A=12244509300en_US
dc.identifier.scopusauthoridZhang, X=8606600100en_US
dc.identifier.scopusauthoridNewsted, JL=6603677236en_US
dc.identifier.scopusauthoridJones, PD=34771015600en_US
dc.identifier.scopusauthoridAu, D=7004909228en_US
dc.identifier.scopusauthoridKong, R=7005290687en_US
dc.identifier.scopusauthoridWu, RSS=7402945079en_US
dc.identifier.scopusauthoridGiesy, JP=35459135300en_US
dc.identifier.scopusauthoridHecker, M=35247848500en_US

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