Juergens K et al. (2005), Isolation and comparative expression analysis o...

XB-ART-1719

Dev Dyn 2005 Aug 01;2334:1554-9. doi: 10.1002/dvdy.20470.

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Isolation and comparative expression analysis of the Myc-regulatory proteins Mad1, Mad3, and Mnt during Xenopus development.

Juergens K , Rust B , Pieler T , Henningfeld KA .

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The Myc-Max-Mad network of transcription factors plays an essential role in many cellular processes such as proliferation, differentiation, and apoptosis. The Mad proteins heterodimerize with Max, function as transcriptional repressors, and are capable of antagonizing the transforming activity of Myc. We report on the isolation of Xmad1, Xmad3, and Xmnt, novel Xenopus genes belonging to the Mad family. We also describe their temporal and spatial expression patterns during Xenopus embryogenesis. Xmad1 expression is found primarily in cells that have undergone terminal differentiation including the notochord, floor plate, and cement gland. Xmad3 transcripts are expressed broadly throughout the central nervous system and the eye, starting at neurula stages. In contrast, Xmnt expression in the CNS was localized anteriorly and, in addition, is present in the migrating neural crest cells. This study demonstrates the Mads are expressed in specific and mostly nonoverlapping patterns, suggesting distinct roles during embryogenesis.

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Species referenced: Xenopus laevis
Genes referenced: h4c4 lat mnt mxd1 mxd3 myc nppa smad1

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	Figure 3. Spatial expression pattern of Xmad1, Xmad3, and Xmnt in Xenopus embryos. (A-I) Xmad1, (J-P) Xmad3, and (Q-W) Xmnt. (A) Stage 6.5 lat., (B) stage 10 veg., (C) stage 11 veg., (D) stage 13 d., (E) stage 17 ant., (F) stage 17, transversal section, (G) stage 19, (H) stage 27, (I) stage 27, transversal section, (J) stage 20 d., (K) stage 20 lat., (L) stage 23 ant., (M) stage 23 d., (N) stage 29, (O) transversal section S1 of N, (P) transversal section S2 of N, (Q) stage 19 ant., (R) stage 19 d., (S) stage 20 lat., (T) stage 23 lat., (U) stage 27, (V) stage 27 sagittal section S1, and (W) stage 27 transversal section S2. anp, anterior neural plate; b, branchial crest segment; cg, cement gland; d, dorsal; ev, eye vesicle; fb, forebrain; fp, floor plate; h, hyoid crest segment; hb, hindbrain; hc, hypochord; lat, lateral; mz marginal zone; nc, notochord; nt, neural tube; veg, vegetal; mb, midbrain; op, olfactory placode; otic vesicle; P, pronephros; pg, pineal gland; sc, spinal chord; r, retina; V, trigeminal ganglion; VII, geniculate ganglion. The dashed line indicates the plane of the section.
	mxd1 (MAX dimerization protein 1) gene expression gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 27, lateral view, anterior left, dorsal up, head and anterior trunk region.
	mxd1 (MAX dimerization protein 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 13, dorsal view, anterior left.
	mxd1 (MAX dimerization protein 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 17, transverse section mid-trunk region, dorsal up.
	mxd1 (MAX dimerization protein 1) gene expression gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19, lateral view, anterior left, dorsal up.
	mxd1 (MAX dimerization protein 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 27, transverse section mid-trunk region, dorsal up.
	Figure 2. Temporal expression of Xmad1, Xmad3, and XMnt mRNA determined by RT-PCR at various stages of Xenopus development. The ubiquitous marker Histone H4 served as a control.
	Figure 1. Amino acid alignment of the predicted open reading frame (ORF) of Xmad1, Xmad3, and Xmnt (GenBank accession numbers: AY964104, AY964105, and AY964106) with the corresponding mouse sequences. A: Comparison of Mad1 and Mad3 protein. B: Comparison of Mnt sequences. The SID repressor domain, the bHLH domain, and the leucine zipper domain are indicated by green, red, and blue bars, respectively. The overall amino acid identity shared between the Xenopus and mouse sequences was: Mad1 (72%), Mad3 (60.4%), and Mnt (55.7%); within the SID domain: Mad1 (95%), Mad3 (90%), and Mnt (80%) and within the bHLH domain Mad1 (92%), Mad3 (70%), and Mnt (100%).
	mxd3 (MAX dimerization protein 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 20, dorsal view, anterior left.
	mxd3 (MAX dimerization protein 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 20, lateral view, anterior left, dorsal up.
	mxd3 (MAX dimerization protein 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 23, anterior view, dorsal up.
	mxd3 (MAX dimerization protein 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 23, dorsal view, anterior left.
	mxd3 (MAX dimerization protein 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 29, lateral view, anterior left, dorsal up.
	mxd3 (MAX dimerization protein 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 29, transverse section. head region, dorsal up.

	mnt (MAX network transcriptional repressor) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19, anterior view, dorsal up.
	xmt MAX network transcriptional repressor) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 19, dorsal view, anterior down.
	mnt (MAX network transcriptional repressor ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 27, lateral view, anterior left, dorsal up.
	mnt (MAX network transcriptional repressor ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 27, sagittal section through head region, anterior left.
	xmt (MAX network transcriptional repressor ) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 27, transverse section, through spinal chord at pharyngeal level, dorsal up.