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Xenopus ZFP36L1 (zinc finger protein 36, C3H type-like 1) belongs to the ZFP36 family of RNA-binding proteins, which contains two characteristic tandem CCCH-type zinc-finger domains. The ZFP36 proteins can bind AU-rich elements in 3' untranslated regions of target mRNAs and promote their turnover. However, the expression and role of ZFP36 genes during neural development in Xenopus embryos remains largely unknown. The present study showed that Xenopus ZFP36L1 was expressed at the dorsal part of the forebrain, forebrain-midbrain boundary, and midbrain-hindbrain boundary from late neurula stages to tadpole stages of embryonic development. Overexpression of XZFP36L1 in Xenopus embryos inhibited neural induction and differentiation, leading to severe neural tube defects. The function of XZP36L1 requires both its zinc finger and C terminal domains, which also affect its subcellular localization. These results suggest that XZFP36L1 is likely involved in neural development in Xenopus and might play an important role in post-transcriptional regulation.
Figure 1 Bayesian tree depicting the phylogenetic relationships of the ZFP36 proteins. Branch confidence values are shown as
Bayesian posterior probabilities. The scale bar shows the number of substitutions per site.
Figure 2 Temporal and spatial expression patterns of XZFP36L1
A: RT-PCR analysis of the developmental expression of XZFP36L1A and B; B–I: Expression patterns of XZFP36L1 revealed by WMISH and paraffin sections;
B: Stage 10, the embryo was bisected, the arrowheads indicate the expression of XZFP36L1 in the ectoderm and involuting mesoderm; C: Stage 16, anterior
view; D: Stage 20, lateral view; E: Stage 20, anterior view; F-I: Stage 32 embryos: F: Lateral view. The broken lines indicate the position of corresponding
sections showed in F’ and Fâ€. Br, branchial arches. F’, Fâ€: Transverse sections of stage 32 embryo, the arrowheads indicate the expression of XZFP36L1 in the
dorsal brain. ot, otic vesicle; G: Dorsal view of the head region; H: Horizontal section showing its expression in the fore-midbrain boundary (red arrowheads)
and mid-hindbrain boundary (black arrowheads); I: Double WMISH of En2 (red) and XZFP36L1 (blue). Arrowheads in (C−I) indicate the expression of
XZFP36L1 in the forebrain (blue), fore-midbrain boundary (red) and mid-hindbrain boundary (black). White arrowheads in (C) and (E) indicate its expression
at the presumptive neural placode region. Bars in (B−I)=0.5 mm.
Figure 3 Overexpression of XZFP36L1 leads to NTDs in Xenopus embryos
A: Control embryos, stage25; B: Embryo injected with XZFP36L1 mRNA shows severe defects during neural tube closure (arrowheads), stage 25; C−J:
Overexpression of XZFP36L1 inhibits neural differentiation. Expression of neural marker genes N-tubulin (C), Sox2 (D), Slug (E), En2 (F), Nkx6.2 (G), Pax3
(H), Dbx1 (I) and Xath1 (G) is largely inhibited in the injected sides (arrowheads). (C−F), stage 15; (G−J), stage 25. Bars=0.5 mm.
Figure 4 Activity and cellular localization of different XZFP36L1 truncates
A: The schematic structures of XZFP36L1 protein and its truncates; B-G: Both the ZF motif and the C-terminal domain are required for its activity in neural
development; B: control embryos. Overexpression of full length XZFP36L1 (C) or the ZFC truncate (F) leads to defects in neural tube closure (arrowheads),
while overexpression of the NZF truncate (D) or ZF domains (E) or C-terminal truncate (G) has no obvious effect on neural tube closure; H-M: Localization of
XZFP36L1 and its truncates in transfected Hela cells.
Image taken from "Temporal and Spatial Expression Patterns of Sox1 Gene in Xenopus laevis Embryo" which has no PMID and can't be uploaded to Xenbase. Appended to this article as it is the same author and publication
http://www.zoores.ac.cn/EN/Y2007/V28/I4/403#1
Figure1. Alignment of Xenopus laevis Sox1, Sox2, and Sox3
Image taken from "Temporal and Spatial Expression Patterns of Sox1 Gene in Xenopus laevis Embryo" which has no PMID and can't be uploaded to Xenbase. Appended to this article as it is the same author and publication
http://www.zoores.ac.cn/EN/Y2007/V28/I4/403#1
Fig. 2 Temporal expression of Sox1-3 gene
Image taken from "Temporal and Spatial Expression Patterns of Sox1 Gene in Xenopus laevis Embryo" which has no PMID and can't be uploaded to Xenbase. Appended to this article as it is the same author and publication
http://www.zoores.ac.cn/EN/Y2007/V28/I4/403#1
Fig.3 Embryonic expression of Xenopus laevis Sox1 Gene
Image taken from "Temporal and Spatial Expression Patterns of Sox1 Gene in Xenopus laevis Embryo" which has no PMID and can't be uploaded to Xenbase. Appended to this article as it is the same author and publication
http://www.zoores.ac.cn/EN/Y2007/V28/I4/403#1
Fig.4 Comparison of the expression of XSox1-3 in the brain and eye
