title: Detection of mRNA expression patterns by nonradioactive in situ hybridization on histological sections of floral tissue.
abstract: Analysis of gene activity with high spatial resolution is a prerequisite for deciphering regulatory networks which underlie developmental programs. Over many years, in situ hybridization has become the gold standard for the identification of in vivo expression patterns of endogenous mRNAs. Nonetheless, the method has several limitations, and the detection of lowly expressed transcripts is still a challenge. Here, we present a robust protocol for sensitive analysis of expression patterns in inflorescence tissue of Arabidopsis thaliana. We describe how the samples are fixed, embedded, and sectioned in preparation for in situ hybridization, how RNA probes are prepared, and how hybridization and detection is carried out. While the described protocol is optimized for inflorescence meristems, it can possibly be used for other tissues as well.
authors.affiliation: Department of Stem Cell Biology, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Medzihradszky
authors.forename: Kay
authors.initials: K
authors.lastname: Schneitz
authors.forename: Jan U
authors.initials: JU
authors.lastname: Lohmann
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics; D035264:Flowers / Q000166:cytology* / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D006652:Histological Techniques / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods*; D015347:RNA Probes / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016610:Tissue Embedding; D016707:Tissue Fixation; D014618:Vacuum
chemical_list: D015347:RNA Probes; D012333:RNA, Messenger