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Biochem Biophys Res Commun
2009 Jan 16;3783:428-32. doi: 10.1016/j.bbrc.2008.11.069.
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Trim36/Haprin plays a critical role in the arrangement of somites during Xenopus embryogenesis.
Yoshigai E
,
Kawamura S
,
Kuhara S
,
Tashiro K
.
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The TRIM family members contain a tripartite motif (TRIM), which includes RING, B-box, and coiled-coil domains, or collectively RBCC. They have been implicated in a variety of biological processes, such as the regulation of differentiation and development, and oncogenesis. In this study, we discovered a novel function of the TRIM family in early development. We report the expression of Trim36/Haprin during Xenopus laevis early embryogenesis and its involvement in somite formation. Temporal expression analysis indicated that Trim36/Haprin was present throughout embryogenesis. Spatial expression analysis showed that its expression was mainly confined to the nervous system and a portion of the posterior somite. Morpholino-mediated knockdown of Trim36/Haprin markedly and specifically inhibited the somite formation. We conclude that Trim36/Haprin plays an important role in the arrangement of somites during their formation.
Fig. 1. Comparison of the TRIM36/HAPRIN amino acid sequence among species. Predicted TRIM36/HAPRIN amino acid sequences of X. laevis (GenBank Accession No.
BC068723), humans (TRIM36, GenBank Accession No. NM_018700), mice (HAPRIN, GenBank Accession No. AB103063; TRIM36, GenBank Accession No. BC068106), and X.
tropicalis (TRIM36, GenBank Accession No. CR942777) are compared. RING finger (RING), B-box 1 and 2 type zinc finger (B1 and B2), coiled-coil (CC), fibronectin type III (FN3),
and SPRY domains are presented. The digits indicate the amino acid residues that define the boundaries of the domains.
Fig. 2. Temporal and spatial expression pattern of X. laevis Trim36/Haprin during development. (A) RT-PCR analysis during X. laevis embryogenesis. The amount of input cDNA
was confirmed by PCR with ornithine decarboxylase primers (ODC). (B) RT-PCR analysis using each blastomere at the 8-cell embryos. Abbreviations: animal, animal pole cell;
vegetal, vegetal pole cell; dorsal, dorsal side cell; ventral, ventral side cell. (CâG) Whole-mount in situ hybridization analysis showed the localization of X. laevis Trim36/Haprin
transcripts. Dorsal views at stage 18 (C) are shown with the anterior region towards the bottom. Lateral views at stage 23 (D) and 28 (E) are shown with the anterior region
towards the left. Transverse sections at stage 28 at the head (F) and the posterior regions (G) are also shown. The positions of the sections are indicated in (E) as vertical lines.
Black arrows, white arrows and arrowheads indicate somites, eyes, and neural tubes, respectively.
Fig. 3. Effect of Trim36/Haprin MO microinjection. (A) Embryos microinjected with
Trim36/Haprin MO. Trim36/Haprin MO at a concentration of 10 or 20 ng was injected
into 1-cell embryos (1-cell) and both blastomeres of 2-cell embryos (right and left).
(B) The frequency of malformation is shown in (A).
Fig. 4. Histological and morphological analyses. (A) Whole-mount in situ hybridization with a DIG-labeled antisense a-actin RNA probe using the embryo injected with
Trim36/Haprin MO at the left blastomere in the 2-cell stage. (B) The histological analysis of Trim36/Haprin MO-microinjected (MO), and MO and Trim36/Haprin mRNA-coinjected
(MO + mRNA) embryos. These embryos were injected into all four blastomeres of the 8-cell vegetal side.
trim36 (tripartite motif containing 36) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 28, lateral view, anteriorleft, dorsal up.
