Zhong C et al. (2015), Developmental expression of the N-myc downstrea...

XB-ART-51841

Int J Dev Biol 2015 Jan 01;5910-12:511-7. doi: 10.1387/ijdb.150178xh.

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Developmental expression of the N-myc downstream regulated gene (Ndrg) family during Xenopus tropicalis embryogenesis.

Zhong C , Zhou YK , Yang SS , Zhao JF , Zhu XL , Chen HH , Chen PC , Huang LQ , Huang X .

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The N-myc downstream regulated gene (Ndrg) family consists of four main members Ndrg1, 2, 3, and 4. The Ndrg genes are involved in many vital biological events including development. However, comprehensive expression patterns of this gene family during vertebrate embryogenesis remain largely unknown. Here, we analyzed the Ndrg family from the evolutionary perspective and examined the expression patterns of the Ndrg genes during Xenopus tropicalis embryogenesis. Different Ndrg family members of vertebrates are separated into different homology clusters which can be further classified into two groups and each Ndrg family member is well conserved during evolution. The temporal and spatial expression patterns of Ndrg1, 2, 3 and 4 are different during early Xenopus tropicalis development. Ndrg1, 2 and 4 are maternally expressed genes while Ndrg3 is a zygotically expressed gene. The Ndrg genes are differentially expressed in the developing central nervous system, the developing sensory organs, and the developing excretory organs. Moreover, they also show other specific expression domains. Our results indicate that the Ndrg genes exhibit specific expression patterns and may play different roles during vertebrate embryogenesis.

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Species referenced: Xenopus tropicalis
Genes referenced: myc mycn ndrg1 ndrg2 ndrg3 ndrg4 ptpru

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	Fig. 1. Amino acid sequences alignment of the Ndrg genes. (A-D) Amino acid sequences of Ndrg1, 2, 3, and 4 from different vertebrates were aligned respectively. Amino acid sequences were collected from human (Homo sapiens), mouse (Mus musculus), chicken (Gallus gallus), frog (Xenopus tropicalis), and zebrafish (Danio rerio). Identity residues from zebrafish to mammals are shown in black, and similarity sites are shown in gray. The red, blue, green and pink lines indicate Ndr domain, a/b hydrolase-fold motif, phosphopantetheine site and 3×10 AA (GTRSRSHTSE) tandem repeats, respectively. Abbreviations: hs (Homo sapiens), mm (Mus musculus), gg, (Gallus gallus), xt (Xenopus tropicalis), dr (Danio rerio).
	Fig. 2. Phylogenetic tree of the Ndrg genes. The CDSs of the Ndrg genes were obtained from human (Homo sapiens), chimpanzee (Pan troglodytes), gibbion (Nomascus leucogenys), cow (Bos taurus), rat (Rattus norvegicus), mouse (Mus musculus), opossum (Monodelphis domestica), chicken (Gallus gallus), frog (Xenopus tropicalis), zebrafish (Danio rerio), fruit fly (Drosophila melanogaster), and worm (Caenorhabditis elegans). NJ based phylogenetic tree was constructed by MEGA5 software. Numbers on branch nodes are bootstrap replication values. The scale bar represents the number of amino acids substitutions.
	Fig. 3. The temporal expression patterns of Ndrg1, 2, 3 and 4 during Xenopus tropicalis embryogenesis. qRT-PCR analysis of relative expression levels of Ndrg1, 2, 3 and 4 at eggs (stage 0), cleavage stage (stage 3 and 6), gastrula stage (stage 10), neurula stage (stage 14 and 17), early tailbud stage (stage 23 and 28) and late tailbud stage (stage 34).
	Fig. 4. The spatial expression pattern of Ndrg1 during Xenopus tropicalis embryogenesis. (A) Animal view. (B) Vegetal view, dorsal to the top. (C) Dorsal view, vegetal to the bottom. (D, E) Dorsal view, anterior to the top. (F, G) Lateral view, dorsal to the top and anterior to the right. (H-J) Transverse paraffin sections of stage 28 embryos after WISH. Abbreviations: ba (branchial arch), de (dorsal endoderm), fb (forebrain), em (endomesoderm), nc (neural crest), md (midline), ntc(notochord), opv (optic vesicle), pn (pronephros), vp (ventral pancreatic bud).
	Fig.5. The spatial expression pattern of Ndrg2 during Xenopus tropicalis embryogenesis. (A,B) Animal view. (C,D) Dorsal view, anterior to the top. (E,G) Lateral view, dorsal to the top and anterior to the right. (F) Anterior view, dorsal to the top. (H-K) Transverse paraffin sections of stage 28 embryos after WISH. Abbreviations: br (brain), ep (epidermis), nf (neural fold), nt (neural tube), opv (optic vesicle), otv (otic vesicle), sc (spinal cord).
	Fig. 6 (Left). The spatial expression pattern of Ndrg3 during Xenopus tropicalis embryogenesis. (A,B) Animal view. (C,E) Lateral view, dorsal to the top and anterior to the right. (D) Anterior view, dorsal to the top. (F-H) Transverse paraffin sections of stage 28 embryos after WISH. Abbreviations: br (brain), h (heart), ptp (profundal and trigeminal placodes/ganglia), otv (otic vesicle), sc (spinal cord).
	Fig. 7 (Right). The spatial expression pattern of Ndrg4 during Xenopus tropicalis embryogenesis. (A,B) Animal view. (C,D) Dorsal view, anterior to the top. (E) Anterior view, dorsal to the top. (F, G) Lateral view, dorsal to the top and anterior to the right. (H-J) Transverse paraffin sections of stage 28 embryos after WISH. Abbreviations: ba (branchial arch), fb (forebrain), mb(midbrain), hb (hindbrain), opv (optic vesicle), otv (otic vesicle), sc (spinal cord), pn (pronephros).
	ndrg1 (N-myc downstream regulated 1) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 3, animal view.
	ndrg2 (N-myc downstream regulated 2) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 5, animal view.
	ndrg2 (N-myc downstream regulated 2) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 19, dorsal view, anterior up.
	ndrg2 (N-myc downstream regulated 2) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 24, anterior view, dorsal up.
	ndrg4 (NDRG family member 4) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 23, lateral view, anterior right, dorsal up.
	ndrg3 (N-myc downstream regulated 3) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior right, dorsal up.
	ndrg3 (NDRG family member 3) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 6, animal view.
	ndrg3 (NDRG family member 3) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 23, anterior view, dorsal up.
	ndrg3 (NDRG family member 3) gene expression in Xenopus tropicalis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anterior right, dorsal up.