Biomedical subjects

Shigeo Takeuchi

Publications and source records attributed to Shigeo Takeuchi.

4 recordsLinked to original sources

PubMed2006

Subtilisin-like proprotein convertase activity is necessary for left-right axis determination in Xenopus neurula embryos.

Signaling by members of TGF-beta superfamily requires the activity of a family of site-specific endopeptidases, known as Subtilisin-like proprotein convertases (SPCs), which cleave these ligands into mature, active forms. To explore the role of SPCs in lateral plate mesoderm (LPM) differentiation in Xenopus, two SPC inhibitors, decanoyl-Arg-Val-Lys-Arg-chloromethylketone (Dec-RVKR-CMK) and hexa-arginine, were injected into the left and right LPM of Xenopus neurulae. Left-side injection caused heart-specific left-right reversal, and this phenotype was rescued by co-injection of mature Nodal protein. In contrast, right-side injection caused left-right reversal of both the heart and gut. Tailbud embryos were less sensitive to SPC inhibitors than neurula embryos. Injection of inhibitors into either side of neurula embryos completely abolished expression of the left-LPM-specific genes, Xnr-1, antivin, and pitx2. SPC1 enzyme (Furin) was injected into the left or right LPM of mid-neurula embryos to determine the effect of enhancing SPC activity. Left-side injection of SPC1 did not cause a significant left-right reversal of the internal organs. However, right-side injection of SPC1 strongly induced the expression of Xnr-1 and pitx2 in the right LPM, and caused 100% left-right reversal of both the heart and gut. These results suggest that moderate level of SPC activity in the right LPM of the neurulae is necessary for proper left-right specification. Taken together, SPC enzymatic activity must be present in both LPMs for expression of the left-handed genes and left-right axis determination of the heart and gut in Xenopus embryos.

Anal Canal↗

PubMed2005

Xenopus nodal related-1 is indispensable only for left-right axis determination.

In Xenopus, multiple nodal-related genes are expressed in the organizer region. Among them, only Xenopus nodal related-1 (Xnr-1) is expressed unilaterally in the left lateral plate mesoderm (LPM) at late neurula-early tailbud stage. To elucidate the essential role of Xnr-1 for left-right specification, loss of function experiments using antisense morpholino oligonucleotides (MOs) targeting three different regions of Xnr-1 were performed. Left-side injection of Xnr-1 MO suppresses the left-side specific genes such as Xnr-1, Xenopus antivin (lefty) and Xenopus pitx2 and randomizes cardiac and visceral left-right orientation. In contrast, paraxial bilateral expression of Xnr-1 along the posterior notochord is not affected by the Xnr-1 MO. In embryos injected with the Xnr-1 MO, morphology of dorsal axial structures is normal and dorsal expression of sonic hedgehog and TGF-beta5 is not changed. Right-side injection of Nodal protein, or polyethyleneimine-based gene transfer of Xnr-1 mRNA in the right LPM induces Xnr-1 and pitx2 in the same side and fully (more than 90%) reverses situs of the internal organs. Left-side injection of Nodal protein restores normal left-right orientation in the embryos that were injected with Xnr-1 MO into the left blastomere and would cause randomization of the left-right axis without the Nodal injection. Taken together, unilateral expression of Xnr-1 in the left LPM directs the orientation of the left-right axis by driving the left-specific gene cascade. Knockdown of Xnr-1 function by the MOs suggests that Xnr-1 is indispensable only for the left-right orientation and dispensable for other embryonic axes probably owing to the redundancy in the function of multiple Xnrs.

Animals↗

PubMed2003

Xenopus neurula left-right asymmetry is respeficied by microinjecting TGF-beta5 protein.

A variety of TGF-beta-related ligands regulate the left-right asymmetry of vertebrates but the involvement of TGF-betas in left-right specification has not been reported. We assessed whether TGF-beta signaling is involved in the left-right specification of Xenopus post-gastrula embryos by microinjecting Xenopus TGF-beta5 protein into the left or right flank of neurula-tailbud embryos. Injection on the right side of neurulae caused left-right reversal of the internal organs in 93% of the embryos, while injection on the left side caused less than 5% left-right reversal. Expression of Xenopus nodal related-1 (Xnr-1 ), Xenopus antivin and Xenopus Pitx2, which are normally expressed on the left, was unaltered by the left-side injection. In contrast, right-side injection into neurulae induced the expression of these genes predominantly on the right side. Right-side injection into tailbud embryos caused bilateral expression of these handed genes. Time course analysis of asymmetric gene expression revealed that Xnr-1 could be induced by TGF-beta5 at late neurula stage, while antivin and Pitx2 could be induced by TGF-beta5 at the latertail bud stage. Injection of the antisense morpholino oligonucleotide against Xenopus TGF-beta5 into the left dorsal blastomere inhibited the normal left-handed expression of Xnr-1 and Pitx2, and caused the organ reversal in the injected embryos. These results suggest that normal left-right balance of endogenous TGF-beta5 signaling in the neurula embryo may be needed to determine the laterality of the asymmetric genes and to generate the correct left-right axis.

Animals↗

PubMed2002

Changes in gustatory sense during pregnancy.

Changes in gustatory sense were investigated in 97 pregnant women and in 30 healthy, non-pregnant women who served as controls. All 97 pregnant women completed a questionnaire regarding taste changes and 32 of them underwent serial tests of gustatory function, including electrogustometry and testing with filter paper disks placed over the areas of the chorda tympani nerve and glossopharyngeal nerve. All gustatory testing was performed by the same person. In addition, serum levels of trace elements were measured in 72 of the pregnant women. Of the 97 pregnant women, 90 (92.8%) reported in the questionnaire that they had experienced some type of change in taste during pregnancy, usually a change in sour taste (59 women; 65.6%). Pregnant women had higher gustatory thresholds than non-pregnant women, with an especially marked decrease in gustatory function being noticed in the first trimester. Although serum zinc levels decreased in pregnant women between the second and third trimesters, zinc levels were in the normal range in the early stage of pregnancy. Thus, it is difficult to explain dysgeusia in the early stage of pregnancy as being associated with a deficiency of zinc. The decrease in gustatory function during the first trimester is considered to be due to the notable changes in secretion of hormones that occur during this stage.

Adolescent↗