Biomedical subjects

Kumiko Yamaguchi

Publications and source records attributed to Kumiko Yamaguchi.

5 recordsLinked to original sources

PubMed2006

Developmental expression of the chick four-jointed homologue.

Four-jointed is a type II transmembrane protein that is thought to be cleaved to give rise to a secreted protein. In Drosophila, four-jointed controls outgrowth, vein patterning, and bristle polarity in the developing limb together with the polarity of the ommatidia in the developing eye. In Drosophila and mice, Fj is regulated by notch signaling. Here, we have determined the expression of the chick four-jointed (fjx) homologue during embryonic development. We show that fjx is expressed in the limb bud; facial primordia; the proliferating zone of the lens, feather buds, the neural tube; and neural crest derivatives such as the dorsal root ganglia. Analysis of the fjx expression in the developing limb bud showed that initially fjx is expressed throughout the limb bud, but as the limb develops, highest levels of fjx transcripts are found distally. However, by stage 27, fjx expression is predominantly found in the central core of the limb bud. Finally, fjx expression becomes confined to the developing tendons, ligaments, articular cartilage, and arteries but not the veins. Comparison with scleraxis (scx), a marker of tendons and ligaments, revealed that they are coexpressed in the majority of tendons but that fjx is expressed after scx, when the tendons have begun to differentiate. These data suggest that fjx has two roles during limb development: the first controlling outgrowth and the second tissue differentiation.

Amino Acid Sequence↗

PubMed2005

Expression and function of Bapx1 during chick limb development.

The homeobox-containing transcription factor Bapx1 (also known as Nkx3.2) is crucial for development of the axial skeleton and parts of the chondrocranium. Here we describe the detailed expression of Bapx1 during chick limb development and show that in contrast to its expression in the axial skeleton, Bapx1 is expressed after the commitment to chondrogenesis. Bapx1 is initially expressed throughout the developing skeletal elements prior to the overt differentiation of the distinct chondrogenic layers. Once distinct layers (proliferating, prehypertrophic and hypertrophic) have formed, Bapx1 expression is restricted to the proliferating chondrocytes. Bapx1 transcripts are excluded from the articular cartilage. A second homeobox-containing transcription factor, Barx1, is expressed in a complementary fashion in the developing joint and articular cartilage. Interestingly, in vitro functional analyses showed that Bapx1 overexpression in micromass cultures increased both matrix production and nodule number suggesting that Bapx1 is sufficient to promote chondrogenesis in the limb. In contrast, Barx1 had the opposite effect on nodule number suggesting that it has an inhibitory effect on chondrogenic initiation consistent with its expression in the developing joint. A slight increase in matrix levels was also observed consistent with its expression in the articular chondrocytes. Finally, we show that Bapx1 is also expressed in the soft tissues such as the developing tendons, muscle sheaths and surrounding mesenchyme, and therefore may have additional as yet uncharacterized roles in limb morphogenesis.

Amino Acid Sequence↗

PubMed2004

Using protein-DNA chimeras to detect and count small numbers of molecules.

We describe general methods to detect and quantify small numbers of specific molecules. We redirected self-splicing protein inteins to create 'tadpoles', chimeric molecules comprised of a protein head covalently coupled to an oligonucleotide tail. We made different classes of tadpoles that bind specific targets, including Bacillus anthracis protective antigen and the enzyme cofactor biotin. We measured the amount of bound target by quantifying DNA tails by T7 RNA polymerase runoff transcription and real-time polymerase chain reaction (PCR) evaluated by rigorous statistical methods. These assays had a dynamic range of detection of more than 11 orders of magnitude and distinguished numbers of molecules that differed by as little as 10%. At their low limit, these assays were used to detect as few as 6,400 protective antigen molecules, 600 biotin molecules and 150 biotinylated protein molecules. In crudely fractionated human serum, the assays were used to detect as few as 32,000 protective antigen molecules. Tadpoles thus enable sensitive detection and precise quantification of molecules other than DNA and RNA.

Antigens, Bacterial↗

PubMed2004

Premature regression of the leg apical ectodermal ridge in the Japanese chick wingless mutant.

The autosomal dominant Japanese wingless mutant has varying degrees of wing and leg truncations. The wing defects range from complete loss to negligible defects, whereas leg abnormalities are usually restricted to loss of the phalanges. Further analyses of the mutant focusing on the leg, which has been relatively uncharacterized, were performed. The expression pattern of Fgf8, a marker gene for the apical ectodermal ridge (AER) that controls outgrowth of the limbs, revealed premature regression at stage 28. Electron microscopy study showed abnormalities in the basement membrane all through the AER in the same stage. In the mutant, cell death was observed in the mesenchyme underlying AER between stages 31 and 32, although in the wild-type leg, AER regression and cell death occurred almost simultaneously at stages 33-34. To know if the cell death and cessation of the outgrowth are common mechanisms of wild-type and the mutant, we removed the AER in wild-type embryos at stage 28 and followed the fate of the limb. This also resulted in premature cell death 48 h after AER removal (equivalent to stage 32) and limb truncations similar to those observed in mutant limbs. To confirm whether either AER or underlying mesenchyme is responsible for the truncation, transplantation of the AER between the wild-type and the mutant was performed. This revealed that AER is the defective tissue in this mutant.

Animals↗

PubMed2004

Spatiotemporal distribution of apoptosis during normal cloacal development in mice.

To understand normal cloacal developmental processes, serial sagittal sections of mouse embryos were made every 6 hrs from embryonic day 11.5 (E11.5) to E13.5. During cloacal development to form the urogenital sinus and anorectal canal, fusion of the urorectal septum with the cloacal membrane was not observed, and the ventral and dorsal parts of the cloaca were continuously connected by the canal until disappearance of the cloacal membrane to open the vestibule formed by the urogenital sinus and anorectal canal to the outside at E13.5. Ventral shifting of the dorsal part of the cloaca was observed until E12.5. The dorsal part was transformed in accordance with ventral shifting. In addition, apoptosis was seen to occur around the dorsal part. However, from E12.25, apoptotic cells are observed in a linear arrangement in the urorectal septum just ventral to the peritoneal cavity. Interestingly, extension of this line reaches the area of the cloacal membrane disintegrated by apoptosis. The present findings suggest that in the early stages (until E12.0), distribution of apoptosis in mesenchyme around the dorsal part of the cloaca might be strongly related to the transformation and ventral shifting of this part. Conversely, the apoptosis pattern in urorectal septum mesenchyme in later stages (from E12.0) might be involved in transformation of the urorectal septum and disintegration of the cloacal membrane.

Aging↗