Whole-mount in situ hybridizationon zebrafish embryos using a mixture of digoxigenin- and fluorescein-labelled probes

The 6-O-endosulfatase enzymes (Sulfs) edit the final sulfation pattern and function of heparan sulfate (HS) by removal of 6-O-sulfate groups from the chain. To date, two mammalian sulf genes have been identified that regulate many signalling pathways during embryonic development. In zebrafish a sulf1 ortholog and duplicate copies of the mammalian sulf2 gene, sulf2a and sulf2, have been identified, which contain conserved motifs characteristic of vertebrate sulf genes. Zebrafish sulf1 and sulf2a are broadly expressed in the central nervous system (CNS) and non-neuronal tissue including heart, somite boundaries, olfactory system, and otic vesicle, whereas sulf2 expression is almost entirely restricted to the CNS. The duplicate copies of sulf2 have distinct expression patterns, which together mirror that of mouse sulf2, suggesting duplication in the teleost lineage has been followed by subfunctionalisation, whereby both genes need to be preserved by selection to ensure the ancestral gene's expression profile and function is maintained. Developmental Dynamics 239:3312-3323,

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“…Sense probes were used to confirm the specificity of antisense probes. WISH were performed using standard methods (Jowett and Lettice, 1994).…”

Section: Whole Mount In Situ Hybridisationmentioning

confidence: 99%

Dynamic expression patterns of 6‐O endosulfatases during zebrafish development suggest a subfunctionalisation event for sulf2

Gorsi

Whelan

2010

Developmental Dynamics

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“…One-colour whole-mount in situ hybridization was performed as described previously (Jowett and Lettice, 1994) with modifications described elsewhere (Hauptmann and Gerster, 1994;Weidinger et al, 2002).…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Transition from non-motile behaviour to directed migration during early PGC development in zebrafish

Blaser

Eisenbeiß

Neumann

et al. 2005

Journal of Cell Science

167

189

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The migration of zebrafish primordial germ cells (PGCs) is directed by SDF-1a and serves as a model for long-range chemokine-guided cell migration. Whereas the development and migration of zebrafish PGCs have been studied in great detail starting at mid-gastrulation stages when the cells exhibit guided active migration [7-8 hours post fertilization (hpf)], earlier stages have not yet been examined. Here we show that the PGCs acquire competence to respond to the chemokine following discrete maturation steps. Using the promoter of the novel gene askopos and RNA elements of nanos1 to drive GFP expression in PGCs, we found that immediately after their specification (about 3 hpf) PGCs exhibit simple cell shape. This stage is followed by a phase at which the cells assume complex morphology yet they neither change their position nor do they respond to SDF-1a. During the third phase, a transition into a `migratory stage' occurs as PGCs become responsive to directional cues provided by somatic cells secreting the chemokine SDF-1a. This transition depends on zygotic transcription and on the function of the RNA-binding protein Dead end and is correlated with down regulation of the cell adhesion molecule E-cadherin. These distinctive morphological and molecular alterations could represent a general occurrence in similar processes critical for development and disease.

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“…Whole-mount in situ hybridization was essentially as described (Oxtoby and Jowett, 1993) and either 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium (BCIP/NBT) or BM purple is used as a chromogenic substrate; two-color in situ hybridization was performed basically as previously described (Hauptmann and Gerster, 1994;Jowett and Lettice, 1994). lfng was linearized with EcoRI and transcribed with T7; mfng was linearized with HindIII and transcribed with T3; rfng was linearized with EcoRI and transcribed with T7; deltaA was linearized with EcoRI and transcribed with T7 (Haddon et al, 1998b); serrateB was linearized with Xba I and transcribed with T7 (Haddon et al, 1998a); and mariposa was linearized with BamHI and transcribed with T7.…”

Section: In Situ Hybridization and Imagingmentioning

confidence: 99%

Sequence and embryonic expression of three zebrafish fringe genes: lunatic fringe, radical fringe, and manic fringe

Qiu

Haddon

et al. 2004

Developmental Dynamics

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Drosophila fringe and its homologues in vertebrates code for glycosyltransferases that modify Notch, altering the sensitivity of this receptor protein to its ligands Delta and Serrate and, thereby, playing an essential part in the demarcation of tissue boundaries. We describe the isolation and characterization of three zebrafish (Danio rerio) fringe homologues: lunatic fringe (lfng), radical fringe (rfng), and manic fringe (mfng). In addition to the sites previously described (Prince et al.

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Whole-mount in situ hybridizationon zebrafish embryos using a mixture of digoxigenin- and fluorescein-labelled probes

Cited by 369 publications

References 2 publications

Dynamic expression patterns of 6‐O endosulfatases during zebrafish development suggest a subfunctionalisation event for sulf2

Dynamic expression patterns of 6‐O endosulfatases during zebrafish development suggest a subfunctionalisation event for sulf2

Transition from non-motile behaviour to directed migration during early PGC development in zebrafish

Sequence and embryonic expression of three zebrafish fringe genes: lunatic fringe, radical fringe, and manic fringe

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