El-Hodiri HM and Kelly LE (2018), Visualization of Gene Expression Patterns by In...

XB-ART-55021

Methods Mol Biol 2018 Jan 01;1797:325-335. doi: 10.1007/978-1-4939-7883-0_17.

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Visualization of Gene Expression Patterns by In Situ Hybridization on Early Stages of Development of Xenopus laevis.

El-Hodiri HM , Kelly LE .

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In situ hybridization performed using whole fixed embryos provides accurate and detailed visualization of gene expression patterns. These patterns are useful for investigating spatial patterns of gene expression in normally developing embryos but can also be useful in investigating the effects of genetic or environmental changes on expression of genetic markers characteristic of particular tissues, organs, or genetic pathways. Our lab's protocol for whole-mount in situ hybridization is presented.

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Species referenced: Xenopus laevis
Genes referenced: alx1 alx4 cyp26a1 prrx1 prrx2

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	Fig. 1 Visualization of paired-type homeobox gene expression in the developing head skeleton of tailbud stage Xenopus laevis embryos. Whole-mount in situ hybridization was performed on embryos fixed at stages 28–30 (panels a–d) or 34–36 (panels e–h) using antisense riboprobes for alx1 (a, e), alx4 (b, f), prrx1 (c, g), or prrx2 (d, h). Specific stages are indicated in the lower right corner of each panel. Gene-specific antisense riboprobes were synthesized by in vitro transcription (Ambion MEGAscript kit) using specific cDNAs generated by 3′-RACE (Clontech SMARTer RACE Kit) using total RNA isolated from mixed neurula stage embryos using degenerate primers encoding VQVWFQN (5′-GTN CAR GTN TGG TTY CAR AAY-3′). Anatomical features are indicated in panel G: C—cement gland; E—eye; F—frontonasal process; Op—olfactory placode/pit; Ov—otic vesicle; P—pharyngeal arch


	alx4 (ALX homeobox 4) gene expression in Xenopus laevis embryo, head region only, assayed via in situ hybridization, NF stage 28, lateral view, anterior left, dorsal up.