Biomedical subjects

Hiroyuki Ide

Publications and source records attributed to Hiroyuki Ide.

15 recordsLinked to original sources

PubMed2006

Limb regeneration in Xenopus laevis froglet.

Limb regeneration in amphibians is a representative process of epimorphosis. This type of organ regeneration, in which a mass of undifferentiated cells referred to as the "blastema" proliferate to restore the lost part of the amputated organ, is distinct from morphallaxis as observed, for instance, in Hydra, in which rearrangement of pre-existing cells and tissues mainly contribute to regeneration. In contrast to complete limb regeneration in urodele amphibians, limb regeneration in Xenopus, an anuran amphibian, is restricted. In this review of some aspects regarding adult limb regeneration in Xenopus laevis, we suggest that limb regeneration in adult Xenopus, which is pattern/tissue deficient, also represents epimorphosis.

Animals↗

PubMed2006

Molecular characterization of Ciona sperm outer arm dynein reveals multiple components related to outer arm docking complex protein 2.

Using proteomic and immunochemical techniques, we have identified the light and intermediate chains (IC) of outer arm dynein from sperm axonemes of the ascidian Ciona intestinalis. Ciona outer arm dynein contains six light chains (LC) including a leucine-rich repeat protein, Tctex1- and Tctex2-related proteins, a protein similar to Drosophila roadblock and two components related to Chlamydomonas LC8. No LC with thioredoxin domains is included in Ciona outer arm dynein. Among the five ICs in Ciona, three are orthologs of those in sea urchin dynein: two are WD-repeat proteins and the third one, unique to metazoan sperm flagella, contains both thioredoxin and nucleoside diphosphate kinase modules. The remaining two Ciona ICs have extensive coiled coil structure and show sequence similarity to outer arm dynein docking complex protein 2 (DC2) that was first identified in Chlamydomonas flagella. We recently identified a third DC2-like protein with coiled coil structure, Ci-Axp66.0 that is also associated in substoichiometric amounts with Ciona outer arm dynein. In addition, Oda5p, a component of an additional complex required for assembly of outer arm dynein in Chlamydomonas flagella, also groups with this family of DC2-like proteins. Thus, the assembly of outer arm dynein onto doublet microtubules involves multiple coiled-coil proteins related to DC2.

Animals↗

PubMed2006

Analysis of scleraxis and dermo-1 genes in a regenerating limb of Xenopus laevis.

Xenopus laevis larvae can regenerate an exact replica of the missing part of a limb after amputation at an early limb bud stage. However, this regenerative capacity gradually decreases during metamorphosis, and a froglet is only able to regenerate hypomorphic cartilage, resulting in a spike-like structure (spike). It has been reported that the spike has tissue deformities, e.g., a muscleless structure. However, our previous study demonstrated that the muscleless feature of the spike can be improved. The existence of other kinds of tissue, such as tendon, has not been clarified. In this study, we focused on the tendon and dermis, and we isolated the scleraxis and dermo-1 genes, which are known to be marker genes for the tendon and dermis, respectively. The expressions of these genes were investigated in both the developmental and regenerating processes of a Xenopus limb. Although muscle was needed to maintain scleraxis expression, scleraxis transcription was detectable in the muscleless spike. Additionally, although grafting of matured skin, including dermal tissue, inhibited limb regeneration, the expression of dermo-1, a dermal marker gene, was detected from the early stage of the froglet blastema. These results indicate that tendon precursor cells and dermal cells exist in the regenerating froglet blastema. Our results support the idea that spike formation in postmetamorphic Xenopus limbs is epimorphic regeneration.

Amino Acid Sequence↗

PubMed2006

Characterization of Xenopus digits and regenerated limbs of the froglet.

Xenopus has 4 and 5 digits in a forelimb and hindlimb, respectively. It is thought that their limbs and digits develop in Xenopus by mechanisms that are almost conserved from amphibians to higher vertebrates. This is supported by some molecular evidence. The 5'hoxd genes are convenient marker genes for characterizing digits in the chick and mouse. The anteriormost digit is characterized by being hoxd13-positive and hoxd12 (hoxd11)-negative in the chick and mouse. In this study, we revealed that the anteriormost digit of the Xenopus forelimb is hoxd13-positive and hoxd11-positive, that is, a more posterior character than digit I. The order of formation of digit cartilages also suggested that Xenopus forelimb digit identity is II to V, not I to IV. We have also been interested in the relationship between digit identity and shh. The anteriormost digit develops in a shh-independent way. A limb treated with cyclopamine (a shh inhibitor) has a gene expression pattern (hoxd11-negative) similar to that in shh-deficient mice, suggesting that a hindlimb treated with cyclopamine has a digit I character. However, a Xenopus froglet regenerate (spike), which lacks shh expression during its regeneration process, does not have such an expression pattern, being hoxd11-positive. We investigated hoxd11 transcriptions in blastemas that formed in the anteriormost and posteriormost digits, and we found that the blastemas have different hoxd11 expression levels. These findings suggest that the froglet limb blastema does not have a mere digit I character in spite of shh defectiveness and that the froglet limb blastema recognizes its positional differences along the anterior-posterior axis.

Amino Acid Sequence↗

PubMed2005

Nerve-dependent and -independent events in blastema formation during Xenopus froglet limb regeneration.

Blastema formation, the initial stage of epimorphic limb regeneration in amphibians, is an essential process to produce regenerates. In our study on nerve dependency of blastema formation, we used forelimb of Xenopus laevis froglets as a system and applied some histological and molecular approaches in order to determine early events during blastema formation. We also investigated the lateral wound healing in comparison to blastema formation in limb regeneration. Our study confirmed at the molecular level that there are nerve-dependent and -independent events during blastema formation after limb amputation, Tbx5 and Prx1, reliable markers of initiation of limb regeneration, that start to be expressed independently of nerve supply, although their expressions cannot be maintained without nerve supply. We also found that cell proliferation activity, cell survival and expression of Fgf8, Fgf10 and Msx1 in the blastema were affected by denervation, suggesting that these events specific for blastema outgrowth are controlled by the nerve supply. Wound healing, which is thought to be categorized into tissue regeneration, shares some nerve-independent events with epimorphic limb regeneration, although the healing process results in simple restoration of wounded tissue. Overall, our results demonstrate that dedifferentiated blastemal cells formed at the initial phase of limb regeneration must enter the nerve-dependent epimorphic phase for further processes, including blastema outgrowth, and that failure of entry results in a simple redifferentiation as tissue regeneration.

Animals↗

PubMed2005

Muscle formation in regenerating Xenopus froglet limb.

A spike, a resultant regenerate made after amputation of a Xenopus froglet limb, has no muscle tissue. This muscle-less phenotype was analyzed by molecular approaches, and the results of analysis revealed that the spike expresses no myosin heavy chain or Pax7, suggesting that neither mature muscle tissue nor satellite cells exist in the spike. The regenerating blastema in the froglet limb lacked some myogenesis-related marker genes, myoD and myf5, but allowed implanted muscle precursor cells to survive and differentiate into myofiber. Implantation of hepatocyte growth factor (HGF) -releasing cell aggregates rescued this muscle-less phenotype and induced muscle regeneration in Xenopus froglet limb regenerates. These results suggest that failure of regeneration of muscle is due to a disturbance of the early steps of myogenesis under a molecular cascade mediated by HGF/c-met. Improvement of muscle regeneration in the Xenopus adult limb that we report here for the first time will give us important insights into epimorphic tissue regeneration in amphibians and other vertebrates.

Amputation, Surgical↗

PubMed2005

Splanchnic (visceral) mesoderm has limb-forming ability according to the position along the rostrocaudal axis in chick embryos.

Positioning of the limb is one of the important events for limb development. In the early stage of embryogenesis, the lateral plate mesoderm splits into two layers and the dorsal layer (the somatic mesoderm) gives rise to a series of distinct structures along the rostrocaudal axis, including the forelimb bud, flank body wall, and hindlimb bud. To determine whether positional information in the somatic mesoderm for regionalization along the rostrocaudal axis is also inherited by the ventral layer of the lateral plate mesoderm (the splanchnic mesoderm), experiments in which the splanchnic mesoderm was transplanted under the ectoderm in an in ovo chick system were carried out. Transplantation of the wing-, flank-, and leg-level splanchnic mesoderm resulted in the formation of wings, nothing, and legs, respectively. These results suggest that the splanchnic mesoderm possesses the ability to form limbs and that the ability differs according to the position along the rostrocaudal axis. The position-specific ability to form limbs suggests that there are some domains involved in the formation of position-specific structures in the digestive tract derived from the splanchnic mesoderm, and results of cell fate tracing supported this possibility. In contrast, analysis of shh expression suggested that the anteroposterior polarity in the limb region seems not to be inherited by the splanchnic mesoderm. We propose that the positioning of limb buds is specified and determined in the very early stage of development of the lateral plate mesoderm before splitting and that the polarity in a limb bud is established after the splitting of the mesoderm.

Animals↗

PubMed2005

Joint development in Xenopus laevis and induction of segmentations in regenerating froglet limb (spike).

In Xenopus laevis, amputation of the adult limb results in the formation of a simple (hypomorphic) spike-like structure without joints, although tadpole limb bud regenerates complete limb pattern. The expression of some joint marker genes was examined in limb development and regeneration. Bmp-4 and gdf-5 were expressed and sox-9 expression was decreased in the joint region. Although developing cartilages were well-organized and had bmp-4 expressing perichondrocytes, the spike cartilage did not have such a structure, but only showed sparse bmp-4 expression. Application of BMP4-soaked beads to the spike led to the induction of a joint-like structure. These results suggest that the lack of joints in the spike is due to the deficiency of the accumulation of the cells that express bmp-4. Improvement of regeneration in the Xenopus adult limb that we report here for the first time will give us important insights into epimorphic regeneration.

Animals↗

PubMed2005

Characteristics of initiation and early events for muscle development in the Xenopus limb bud.

In Xenopus laevis, limb buds start to develop at a later point of the larval stage, prior to metamorphosis. This onset of limb development in Xenopus is totally different from that in amniotes such as birds and mammals, in which limb buds emerge at an early stage of embryogenesis, in parallel with other organogenesis. We investigated limb myogenesis in Xenopus, focusing on myogenic gene expression, myogenic ability of limb bud cells in the early stage, and the origin of myogenic precursor cells in the limb bud. The Xenopus early limb bud contains myoD/cardiac actin-positive and pax3/pax7-negative cells. Interestingly, results of transplantation experiments have revealed that this early limb bud contains myogenic precursor cells. In order to know the contribution of myogenic cells in somites to myogenic precursor cells in the early limb bud, we used a Cre-LoxP system for tracing over a long period. The results of fate tracing for myogenic cells in somites of the Xenopus embryo suggested that early-specified myogenic cells in somites do not contribute to limb muscle in Xenopus. Taken together, the results suggest that limb muscle development in Xenopus has characteristics of initiation and early events distinct from those of other vertebrate clades.

Animals↗

PubMed2004

Molecular characterization of radial spoke subcomplex containing radial spoke protein 3 and heat shock protein 40 in sperm flagella of the ascidian Ciona intestinalis.

Members of the heat-shock protein (HSP)40 regulate the protein folding activity of HSP70 proteins and help the functional specialization of this molecular chaperone system in various types of cellular events. We have recently identified Hsp40 as a component of flagellar axoneme in the ascidian Ciona intestinalis, suggesting a correlation between Hsp40 related chaperone system and flagellar function. In this study, we have found that Ciona 37-kDa Hsp40 is extracted from KCl-treated axonemes with 0.5 M KI solution and comigrates with radial spoke protein (RSP)3 along with several proteins as a complex through gel filtration and ion exchange columns. Peptide mass fingerprinting with matrix-assisted laser desorption ionization/time of flight/mass spectrometry revealed that other proteins in the complex include a homolog of sea urchin spokehead protein (homolog of RSP4/6), a membrane occupation and recognition nexus repeat protein with sequence similarity with meichroacidin, and a functionally unknown 33-kDa protein. A spoke head protein, LRR37, is not included in the complex, suggesting that the complex constructs the stalk of radial spoke. Immunoelectron microscopy indicates that Hsp40 is localized in the distal portion of spoke stalk, possibly at the junction between spoke head and the stalk.

Animals↗

PubMed2003

Ephrin-A2 regulates position-specific cell affinity and is involved in cartilage morphogenesis in the chick limb bud.

In the developing limb bud, mesenchymal cells show position-specific affinity, suggesting that the positional identity of the cells is represented as their surface properties. Since the affinity is regulated by glycosylphosphatidylinositol (GPI)-anchored cell surface proteins, and by EphA4 receptor tyrosine kinase, we hypothesized that the GPI-anchored ligand, the ephrin-A family, also contributes to the affinity. Here, we describe the role of ephrin-A2 in the chick limb bud. Ephrin-A2 protein is uniformly distributed in the limb bud during early limb development. As the limb bud grows, expression of ephrin-A2 is strong in its proximal-to-intermediate regions, but weak distally. The position-dependent expression is maintained in vitro, and is regulated by FGF protein, which is produced in the apical ectodermal ridge. To investigate the role of ephrin-A2 in affinity and in cartilage morphogenesis of limb mesenchyme, we ectopically expressed ephrin-A2 in the limb bud using the retrovirus vector, RCAS. Overexpressed ephrin-A2 modulated the affinity of the mesenchymal cells that differentiate into autopod elements. It also caused malformation of the autopod skeleton and interfered with cartilage nodule formation in vitro without inhibiting chondrogenesis. These results suggest that ephrin-A2 regulates the position-specific affinity of limb mesenchyme and is involved in cartilage pattern formation in the limb.

Animals↗

PubMed2002

Anteroposterior axis formation in Xenopus limb bud recombinants: a model of pattern formation during limb regeneration.

We previously showed that recombinant limb buds with dissociated and reaggregated mesenchyme develop more than 30 digits in Xenopus laevis, which exhibits different capacities for limb regeneration at different developmental stages (Yokoyama et al. [1998] Dev Biol 196:1-10). Cell-cell contact among anterior- and posterior-derived mesenchymal cells is required for anteroposterior (AP) axis formation of recombinant limbs in an intercalary manner. However, whether one-way induction from posterior cells to anterior cells as proposed by the polarizing zone model or interactions between anterior and posterior cells evoke the AP axis formation in recombinant limbs remains unclear. In this study, we found, by a combination of X-ray irradiation and a recombinant limb technique, that not one-way induction but interactions between anterior and posterior cells accompanied by cell contribution are indispensable for AP axis formation in recombinant limbs. Shh was expressed in posterior-derived not anterior-derived cells. We propose that the recombinant limb is an excellent model for examining the axis formation mechanism in regenerating limbs because, as in recombinant limbs, cell-cell contact among cells derived from different positions of an amputation plane occurs in the blastema of regenerating limbs.

Animals↗

PubMed2002

Specification and determination of limb identity: evidence for inhibitory regulation of Tbx gene expression.

Limb-type-specific expression of Tbx5/Tbx4 plays a key role in drawing distinction between a forelimb and a hindlimb. Here, we show insights into specification and determination during commitment of limb-type identity, in particular that median tissues regulate Tbx expressions. By using the RT-PCR technique on chick embryos, the onset of specific Tbx5/Tbx4 expression in the wing/leg region was estimated to be stage 13. Specification of the limb-type identity is thought to occur before stage 9, since all explants from stage 9 through 14 expressed the intrinsic Tbx gene autonomously in a simple culture medium. The results of transplantation experiments revealed that axial structures medial to the lateral plate mesoderm at the level of the wing region are capable of transforming leg identity to wing identity, suggesting that a factor(s) from the median tissues is involved in the limb-type determination. Nevertheless, the transplanted wing region was not converted to leg identity. The results of the transplantation experiments also suggested that wing-type identity is determined much earlier than is leg-type identity. Finally, we also found that inhibitory effects of median tissues mediate the specific expression of Tbx5/Tbx4 in the presumptive wing/leg region. We propose a model for limb-type identification in which inhibitory regulation is involved in restricting one Tbx gene expression by masking the other Tbx expression there.

Animals↗

PubMed2002

Cell adhesiveness and affinity for limb pattern formation.

Stage-dependent cell sorting in vitro is an intriguing property that mesenchymal cells of a chick limb bud have. We previously proposed that N-cadherin, a cell adhesion molecule, is involved in the sorting process and is likely to be a component of the mechanism of proximal-distal patterning in the developing limb (Yajima et al., (1999) Dev. Dynam. 216:274-284). Here, we present more direct evidence that N-cadherin is one of the molecules responsible for regulation of stage-dependent cell sorting in vitro. Our results suggest that N-cadherin, which accumulates in the distal region of the chick limb bud as limb development proceeds, is related to the positional identity that gives rise to the different shapes and numbers of cartilaginous elements along the proximal-distal axis. In this article we also give insights into positional identity which is mediated by Hoxgenes and cell surface property during limb development.

Animals↗

PubMed1998

Freezing behaviors in leaf buds of cold-hardy conifers visualized by NMR microscopy.

1H-Nuclear magnetic resonance (NMR) microscopy was used to study freezing behavior in wintering leaf buds of Momi fir (Abies firma Sieb. et Zucc.) and Japanese red pine (Pinus densiflora Sieb. et Zucc.). The images acquired predominantly reflected the density of mobile (i.e., non-ice) protons mainly from unfrozen water. By comparing images taken at various subfreezing temperatures, we determined which tissues produced the high and low temperature exotherms detected by differential thermal analyses. Typical extra-organ freezing was successfully imaged in leaf buds of A. firma. The bud scales readily froze at -7 degrees C, but shoot primordia remained supercooled to -14 degrees C in December buds and to -21 degrees C in March buds. The size of supercooled shoot primordia was reduced with decreasing temperature, indicating a gradual decrease in water content of the shoot primordia. In contrast, the signal from shoot primordia of P. densiflora disappeared between -7 and -14 degrees C, corresponding to the high temperature exotherm at -8 degrees C, indicating extracellular freezing of the shoot primordia. The xylem and bark tissues readily froze at -7 degrees C in A. firma and between -7 and -14 degrees C in P. densiflora. We conclude that NMR microscopy can noninvasively provide more spatially specific information about freezing behavior in leaf buds than traditional methods such as differential thermal analysis. In particular, it allows the organized and harmonized freezing behaviors in complex organs to be visualized directly thereby revealing the diversity of mechanisms involved in freezing behaviors.

Journal Article↗