Abstract
Xenopus embryos are one of the most used animal models in developmental biology and are well suited for apprehending functions of signaling pathways during embryogenesis. To do so, it is necessary to be able to detect expression pattern of the key genes of these signaling pathways. Here we describe the whole-mount in situ hybridization technique to investigate the expression pattern of ectonucleotidases and purinergic receptors during embryonic development.
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References
Gall JG, Pardue ML (1969) Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A 63:378–383
Tautz D, Pfeifle C (1989) A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma 98:81–85
Hemmati-Brivanlou A, Frank D, Bolce ME, Brown BD, Sive HL, Harland RM (1990) Localization of specific mRNAs in Xenopus embryos by whole-mount in situ hybridization. Development 110:325–330
Harland RM (1991) In situ hybridization: an improved whole-mount method for Xenopus embryos. Methods Cell Biol 36:685–695
Sive HL, Grainger RM, Harland RM (2000) Whole-mount in situ hybridization. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Monsoro-Burq AH (2007) A rapid protocol for whole-mount in situ hybridization on Xenopus embryos. CSH Protoc 2007. https://doi.org/10.1101/pdb.prot4809
Saint-Jeannet JP (2017) Whole-mount in situ hybridization of Xenopus embryos. Cold Spring Harb Protoc. https://doi.org/10.1101/pdb.prot097287
Spemann H, Mangold H (1924) Induction of embryonic primordia by implantation of organizers from different species. In: Willier BH, Oppen-heimer JM (eds) Foundations of experimental embryology. Hafner, New York, pp 144–184
Gould SE, Grainger RM (1997) Neural induction and antero-posterior patterning in the amphibian embryo: past, present and future. Cell Mol Life Sci 53:319–338
Session AM, Uno Y, Kwon T et al (2016) Genome evolution in the allotetraploid frog Xenopus laevis. Nature 538:336–343
Sater AK, Moody SA (2016) Using Xenopus to understand human disease and developmental disorders. Genesis 55. https://doi.org/10.1002/dvg.22997
Tandon P, Conlon F, Furlow JD, Horb ME (2017) Expanding the genetic toolkit in Xenopus: approaches and opportunities for human disease modeling. Dev Biol 426:325–335
Massé K, Eason R, Bhamra S, Dale N, Jones EA (2006) Comparative genomic and expression analysis of the conserved NTPDase gene family in Xenopus. Genomics 87:366–381
Massé K, Bhamra S, Allsop G, Dale N, Jones EA (2010) Ectophosphodiesterase/nucleotide phosphohydrolase (Enpp) nucleotidases: cloning, conservation and developmental restriction. Int J Dev Biol 54:181–193
Blanchard C, Boué-Grabot E, Massé K (2019) Comparative embryonic spatio-temporal expression profile map of the Xenopus P2X receptor family Frontiers in Cellular Neuroscience. https://doi.org/10.3389/fncel.2019.00340
Massé K, Dale N (2012) Purines as potential morphogens during embryonic development. Purinergic Signal 8:503–521
Massé K, Bhamra S, Eason R, Dale N, Jones EA (2007) Purine-mediated signalling triggers eye development. Nature 449:1058–1062
Jowett T, Lettice L (1994) Whole-mount in situ hybridizations on zebrafish embryos using a mixture of digoxigenin- and fluorescein-labelled probes. Trends Genet 10:73–74
Koga M, Kudoh T, Hamada Y, Watanabe M, Kageura H (2007) A new triple staining method for double in situ hybridization in combination with cell lineage tracing in whole-mount Xenopus embryos. Dev Growth Differ 49:635–645
Nieuwkoop F (1994) Normal table of Xenopus laevis (Daudin). Garland Publishing Inc., New York. ISBN: 0-8153-1896-0
David R, Wedlich D (2001) PCR-based RNA probes: a quick and sensitive method to improve whole mount embryo in situ hybridizations. Biotechniques 30:769–772
Oschwald R, Richter K, Grunz H (1991) Localization of a nervous system-specific class II beta-tubulin gene in Xenopus laevis embryos by whole-mount in situ hybridization. Int J Dev Biol 35:399–405
Mizuseki K, Kishi M, Shiota K, Nakanishi S, Sasai Y (1998) SoxD: an essential mediator of induction of anterior neural tissues in Xenopus embryos. Neuron 21:77–85
Penzel R, Oschwald R, Chen Y, Tacke L, Grunz H (1997) Characterization and early embryonic expression of a neural specific transcription factor xSOX3 in Xenopus laevis. Int J Dev Biol 41:667–677
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Blanchard, C., Massé, K. (2020). Developmental Expression of Ectonucleotidase and Purinergic Receptors Detection by Whole-Mount In Situ Hybridization in Xenopus Embryos. In: Pelegrín, P. (eds) Purinergic Signaling. Methods in Molecular Biology, vol 2041. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9717-6_6
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DOI: https://doi.org/10.1007/978-1-4939-9717-6_6
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