How to distinguish between different cell lineages sharing common markers using combinations of double in-situ-hybridization and immunostaining in avian embryos
- Cell migration plays a crucial role in early embryonic development. The chemokine receptor CXCR4 has been reported to guide migration of neural crest cells (NCCs) to form the dorsal root ganglia (DRG) and sympathetic ganglia (SG). CXCR4 also plays an important part during the formation of limb and cloacal muscles. NCCs migration and muscle formation during embryonic development are usually considered separately, although both cell lineages migrate in close neighbourhood and have markers in common. In this study, we present a new method for the simultaneous detection of CXCR4, mesodermal markers and NCCs markers during chicken embryo developmental stages HH18–HH25 by combining double whole-mount in situ hybridization (ISH) and immunostaining on floating vibratome sections. The simultaneous detection of CXCR4 and markers for the mesodermal and neural crest cells in multiple labelling allowed us to compare complex gene expression patterns and it could be easily used for a wide range of gene expression pattern analyses of other chicken embryonic tissues. All steps of the procedure, including the preparation of probes and embryos, prehybridization, hybridization, visualization of the double labelled transcripts and immunostaining, are described in detail.
| Author: | Imadeldin YahyaORCiDGND, Marion BöingGND, Beate Brand-SaberiORCiDGND, Gabriela Morosan-PuopoloORCiDGND |
|---|---|
| URN: | urn:nbn:de:hbz:294-81854 |
| DOI: | https://doi.org/10.1007/s00418-020-01920-7 |
| Parent Title (English): | Histochemistry and cell biology |
| Subtitle (English): | CXCR4-positive mesodermal and neural crest-derived cells |
| Publisher: | Springer Nature |
| Place of publication: | Berlin |
| Document Type: | Article |
| Language: | English |
| Date of Publication (online): | 2021/06/24 |
| Date of first Publication: | 2020/10/10 |
| Publishing Institution: | Ruhr-Universität Bochum, Universitätsbibliothek |
| Tag: | CXCR4; Chicken embryo; Double whole-mount ISH; Immunostaining; Mesodermal cells; NCCs |
| Volume: | 155 |
| First Page: | 145 |
| Last Page: | 155 |
| Note: | Dieser Beitrag ist auf Grund des DEAL-Springer-Vertrages frei zugänglich. |
| Institutes/Facilities: | Institut für Anatomie, Abteilung für Anatomie und Molekulare Embryologie |
| Dewey Decimal Classification: | Technik, Medizin, angewandte Wissenschaften / Medizin, Gesundheit |
| open_access (DINI-Set): | open_access |
| Licence (English): |  Creative Commons - CC BY 4.0 - Attribution 4.0 International |
