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Biochem Biophys Res Commun
2007 Apr 20;3554:902-6. doi: 10.1016/j.bbrc.2007.02.059.
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The mRNA coding for Xenopus glutamate receptor interacting protein 2 (XGRIP2) is maternally transcribed, transported through the late pathway and localized to the germ plasm.
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Using a large-scale in situ hybridization screening, we found that the mRNA coding for Xenopus glutamate receptor interacting protein 2 (XGRIP2) was localized to the germ plasm of Xenopus laevis. The mRNA is maternally transcribed in oocytes and, during maturation, transported to the vegetal germ plasm through the late pathway where VegT and Vg1 mRNAs are transported. In the 3'-untranslated region (UTR) of the mRNA, there are clusters of E2 and VM1 localization motifs that were reported to exist in the mRNAs classified as the late pathway group. With in situ hybridization to the sections of embryos, the signal could be detected in the cytoplasm of migrating presumptive primordial germ cells (pPGCs) until stage 35. At stage 40, when the cells cease to migrate and reach the dorsal mesentery, the signal disappeared. A possible role of XGRIP2 in pPGCs of Xenopus will be discussed.
Fig. 1.
Expression and localization of XGRIP2 mRNA in embryos. We performed in situ hybridization to the sections of embryos at stages 2–6, 10, 19, 28, 35, and 40. Signals can be detected in the germ plasm of blastomeres and cytoplasm of migrating PGCs from stage 10–35. Then, it disappears at stage 40. Arrows indicate the positive signals. Scale bar, 100 μm.
Fig. 2.
Overall expression of XGRIP2 in the ovary and embryos by RT-PCR. The transcript was maternally transcribed and it gradually declined to stage 28, and then increased (because of transcriptional activation in nerve tissues). Control experiment with β-actin was indicated.
Fig. 3.
Expression and localization of XGRIP2 mRNA in oocyte. XGRIP2 mRNA was detected in the cytoplasm of stage-I oocyte and then concentrated to the vegetal region in a wedge shape in stage-II oocyte, and then localized to the germ plasm at stage-III and stage-IV. Scale bar, 100 μm. GV, germinal vesicle; white arrows, mitochondrial cloud.
Supplementary Figure 1.
Amino-acid sequence comparison between XGRIP2 (DDBJ AB290863), predicted Xenopus tropicalis GRIP2 (TGRIP2, Xenbase protein Accession No. 456689) and predicted GRIP1 (TGRIP1, Xenbase protein Accession No. 174390) using the CLUSTALW alignment tool. Consensus sequences are screened; dark blue indicates completely identical, and light blue identical by two to three. Underlined sequences are seven PDZ domains.
Supplementary Figure 2.
Phylogenetic tree was formed by comparison among eleven GRIP family proteins. X, Xenpopus laevis; T, Xenopus tropicalis; C, Canis familiaris; h, human; m, mouse; D, dog; R, rat. Each Accession number is listed in the figure.
