Reza N et al. (2016), Nucleoporin gene expression in Xenopus tropical...

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Int J Dev Biol 2016 Jan 01;604-6:181-8. doi: 10.1387/ijdb.150317nr.

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Nucleoporin gene expression in Xenopus tropicalis embryonic development.

Reza N , Khokha MK , Del Viso F .

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Nucleoporins (nups) compose the structure of the nuclear pore complex (NPC) of all cells, but several studies have illuminated nucleoporins' additional roles in development and the cell cycle. However, a comprehensive study of nup expression in embryonic development has not yet been reported. We synthesized antisense probes for all nup genes and used whole-mount in situ hybridization techniques to determine the expression pattern of all members of the nup family of genes at three different developmental stages in Xenopus tropicalis. We found that the expression of nups was not ubiquitous in embryos, but was localized to specific and distinguishable anatomical structures at all three stages tested. We also found that the expression patterns for nups within the same subcomplexes were not necessarily identical. Thus, nup expression is subject to a significant level of regulation during development. These results provide new information for functional studies of nups to unravel their roles in embryonic development.

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Species referenced: Xenopus tropicalis
Genes referenced: efl1 iqsec1 nup107 nup133 nup153 nup155 nup160 nup188 nup205 nup210 nup214 nup35 nup37 nup43 nup50 nup54 nup62 nup85 nup88 nup93 nup98 rae1 sec13

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	Fig. 3 (Left). Expression of outer ring scaffold nups in developing X. tropicalis embryos. Expression was analyzed by whole mount in situ hybridization at 3 different stages of X. tropicalis development (16-18, 26-28 and 33-36, columns). Signal was detected for all the nups in the anterior regions of the neural tube and near the eye anlages at stages 16-18. At stages 26-28 and 33-36, signal was detected for all nups in the brain, eyes, and visceral arches. c, cement gland; i, isthmus or mid to hindbrain boundary; lm, dorsolateral plate mesoderm; vm, ventral mesoderm. A: anterior, P: posterior, L: left, R: right.
	Fig. 4 (Right). Expression of inner ring scaffold nups in developing X. tropicalis embryos. Expression was analyzed by whole mount in situ hybridization at 3 different stages of X. tropicalis development (16-18, 26-28 and 33-36, columns). All nups were detected in the neural plate region at stages 16-18 and in the head, visceral arches and eyes at stages 26-28 and 33-36. h, heart progenitor field; i, isthmus or mid to hindbrain boundary. A: anterior, P: posterior, L: left, R: right.
	Fig. 5. Expression of Channel nups in developing X. tropicalis embryos. Expression was analyzed by whole mount in situ hybridization at 3 different stages of X. tropicalis development (16-18, 26-28 and 33-36, columns). All nups were detected in neural plate region at stages 16-18; brain, eyes and visceral arches at stages 26-28. i, isthmus or mid to hindbrain boundary. A: anterior, P: posterior, L: left, R: right.
	Fig. 6. Expression of cytoplasmic and nuclear basket nups in developing X. tropicalis embryos. Expression was analyzed by whole mount in situ hybridization at 3 different stages of X. tropicalis development (16-18, 26-28 and 33-36, columns). (A) Cytoplasmic nups and (B) Nuclear basket nups expression patterns. Neural tube signal was detected all nups at stages 16-18. At stages 26-28 and 33-36 transcript expression was detected in the head, eyes, and visceral arches in all nups. h, heart progenitor field; c, cement gland. A: anterior, P: posterior, L: left, R: right.
	Fig. 7. Expression of Transmembrane nups in developing X. tropicalis embryos. Expression was analyzed by whole mount in situ hybridization at 3 different stages of X. tropicalis development (16-18, 26-28 and 33-36, columns). At stages 16-18 expression was detected for all genes in the neural tube. All nups showed expression in the head, eyes, and visceral arches at stages 26-28 and 33-36. A: anterior, P: posterior, L: left, R: right.
	Fig. 8. Correlation of nup expression between in situ hybridization and RTPCR. (A) Schematics of stages 28 and 33 depicting Head (H) and ventral mesoderm (VM) regions that were dissected and used for RNA extractions and subsequent RT-PCR. (B) Expression by RT-PCR of 2 nups for each subcomplex, in each of the dissected tissues. In situ hybridization images from previous figures are shown for illustration. Odc1 (ornithine decarboxylase 1) and ef1a1 (eukaryotic translation elongation factor 1 alpha 1) were used as positive controls of RT-PCR. The primer sequences for each nup are shown in Table 1.