Sánchez RS and Sánchez SS (2013), Characterization of pax1, pax9, and uncx sclero...

XB-ART-46659

Dev Dyn 2013 May 01;2425:572-9. doi: 10.1002/dvdy.23945.

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Characterization of pax1, pax9, and uncx sclerotomal genes during Xenopus laevis embryogenesis.

Sánchez RS , Sánchez SS .

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The axial skeleton develops from the sclerotome, a mesenchymal cell population derived from somites. Sclerotomal cells migrate from somites to the perinotochordal and perineural space where they differentiate into chondrocytes to form cartilage and bone. In anurans, little is known about the way how the sclerotome changes as development proceeds and how these events are regulated at the molecular level. Pax1, Pax9, and Uncx4.1 genes play a central role in the morphogenesis of the axial skeleton in vertebrates, regulating cell proliferation and chondrogenic specification of the sclerotome.In this work, we cloned and examined through whole-mount in situ hybridization and reverse transcriptase-polymerase chain reaction the expression patterns of pax1, pax9, and uncx transcription factors in the anuran Xenopus laevis.We found that these genes are similarly expressed in the sclerotome and in the pharyngeal pouch. A detailed analysis of the location of these transcripts showed that they are expressed in different subdomains of the sclerotomal compartment and differ from that observed in other vertebrates.

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Species referenced: Xenopus laevis
Genes referenced: pax1 pax9 uncx

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	Fig. 4. Spatial expression of pax1 transcripts during Xenopus development. Whole-mount in situ hybridization was carried out using a digoxigenin-labeled riboprobe. A: Lateral view, ante- rior is to the right. A: At stage 22, pax1 is expressed in the branchial arches and weakly in the somite (red arrows). B: The pax1 expression is increased and its localization advances pro- gressively to caudal region of the somites in the stage 24 (B), stage 26 (C), and stage 35 embryos (D). E: Magnification of the dorsal region of D at the somite level. Black arrows indicate the intersomitic space. F: Whole-mount in situ hybridization with pax1 sense probe was used as negative control. G,H: Transverse sections of stage 35 embryos at the branchial arches level (G) and trunk (H). Nieuwenkoop-Faber stages are indicated on the lower left corner. BA, branchial arches; N, notochord; NT, neural tube; PE, pharyngeal endoderm; S, somite.
	pax1 (paired box 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 24, lateral view, anterior left, dorsal up.
	pax1 (paired box 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anterior left, dorsal up.
	pax1 (paired box 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35/36, lateral view, anterior left, dorsal up.
	pax1 (paired box 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35/36, lateral view, anterior left, dorsal up.
	Fig. 5. Spatial expression of pax9 transcripts during Xenopus development. Whole-mount pax9 in situ hybridization. A: Lateral view, anterior is to the right. A: At stage 22, pax9 is weakly expressed in the branchial arches and in the somite (red arrows). B: pax9 expression is increased and its localization advances progressively to caudal region in the stage 26 (B), stage 28 (C), and stage 35 embryos (D). E: Magnification of the dorsal region of C at the somites level. Black arrows indicate the intersomitic space. F: Whole-mount in situ hybridization with pax9 sense probe. G,H: Transverse sections of stage 35 embryos at the branchial arches level (G) and trunk (H).
	pax9 (paired box 9) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anterior left, dorsal up.
	pax9 (paired box 9) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 22, lateral view, anterior left, dorsal up.
	pax9 (paired box 9) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	pax9 (paired box 9) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35/36, lateral view, anterior left, dorsal up.
	pax9 (paired box 9) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35/36, lateral view, anterior left, dorsal up.
	Fig. 6. Spatial expression of uncx transcripts during Xenopus development. Whole-mount uncx in situ hybridization. A: Lateral view, anterior is to the right. A: At stage 21, uncx is weakly expressed in the branchial arches. B: At stage 26, uncx is strongly expressed in the branchial arches and in the anterior somitic region. C,D: Expression uncx in the in stage 28 (C) and stage 35 (D). E: Magnification of the dorsal region of C at the somites level. F: Whole-mount in situ hybridization with pax9 sense probe. G,H: Transverse sections of stage 35 embryos at the bran- chial arches level (G in panel D) and trunk (H in panel D).
	uncx (unc homeobox ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 22, lateral view, anterior left, dorsal up.
	uncx (unc homeobox ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 26, lateral view, anterior left, dorsal up.
	uncx (unc homeobox ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.
	uncx (unc homeobox ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35, lateral view, anterior left, dorsal up.
	uncx (unc homeobox ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 35, lateral view, anterior left, dorsal up.
	Transverse sections of stage 35 embryos at the branchial arches level. Displays pharyngeal endoderm (PE).