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Confocal analysis of hedgehog morphogenetic gradient coupled with fluorescent in situ hybridization of hedgehog target genes
- Source :
- Methods in Molecular Biology (Clifton then Totova), Methods in Molecular Biology (Clifton then Totova), Humana Press (Springer Imprint), 2007, 397, pp.105-14, Methods in Molecular Biology, Methods in Molecular Biology, Humana Press/Springer Imprint, 2007, 397, pp.105-14
- Publication Year :
- 2007
- Publisher :
- HAL CCSD, 2007.
-
Abstract
- Hedgehog (Hh) family members are secreted proteins that can act at short and long range to direct cell fate decisions during developmental processes. In both Drosophila and vertebrates, the morphogenetic gradient of Hh must be tightly regulated for correct patterning. The posttranslational modification of Hh by a cholesterol adduct participates in such regulation. We have shown that cholesterol modification is necessary for the controlled long-range activity of Drosophila Hh, as observed for its vertebrate counterpart Sonic Hh. The presence of cholesterol on Hh allows the observation of large apical punctuate structures of Hh (Hh-LPSs) at a distance from the Hh source both in embryos and in imaginal discs. The Hh-LPSs apical distribution reflects the Hh gradient and is temporally regulated. Hh gradient modulation is directly related to the dynamic expression of the Hh target gene serrate (ser), shown by immunofluorescent detection of Hh coupled with fluorescent in situ hybridization of ser.
- Subjects :
- Embryo, Nonmammalian
Microscopy, Confocal
Staining and Labeling
Genes, Insect
RNA Probes
[ SDV.BC.IC ] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB]
Drosophila melanogaster
[SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB]
Image Processing, Computer-Assisted
Morphogenesis
Animals
Hedgehog Proteins
RNA, Antisense
Digoxigenin
In Situ Hybridization, Fluorescence
Subjects
Details
- Language :
- English
- ISSN :
- 10643745 and 19406029
- Database :
- OpenAIRE
- Journal :
- Methods in Molecular Biology (Clifton then Totova), Methods in Molecular Biology (Clifton then Totova), Humana Press (Springer Imprint), 2007, 397, pp.105-14, Methods in Molecular Biology, Methods in Molecular Biology, Humana Press/Springer Imprint, 2007, 397, pp.105-14
- Accession number :
- edsair.pmid.dedup....84f4f587d50dc9d98deb3fd4406a57b5