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K Muneoka

Publications and source records attributed to K Muneoka.

At least 19 recordsLinked to original sources
PubMed1992

Position specific growth regulation of 3T3 cells in vivo.

In this study we have investigated the mechanism by which spatial growth is regulated by monitoring 3T3 cells, introduced into the developing mouse limb using exo utero surgery. The 3T3 cells were labeled with a human cell surface glycoprotein, CD8, and injected into stage 7-9 mouse limbs. At 24 and 48 hr after injection embryos were labeled with [3H]thymidine and processed for immunohistochemistry and autoradiography. The labeling index of CD8 positive cells was compared to that of neighboring limb bud cells and also to the position of the injection site within the limb. We find that the labeling index of 3T3 cells is in accord with that of the limb cells that immediately surround them; 3T3 cells display a high labeling index in limb regions of high growth and a low labeling index in limb regions of low growth. In addition, we find that both limb bud cells and injected 3T3 cells display a general proximal (low) to distal (high) gradient of growth at the stages analyzed. We conclude from these results that position-specific regulation of growth occurs in a non-cell autonomous manner and is likely to be mediated by mitogenic signals that are localized within the limb environment. In addition, our results demonstrate the usefulness of utilizing established cell lines as in vivo probes to monitor developmental mechanisms.

3T3 Cells↗
PubMed1992

Molecular aspects of regeneration in developing vertebrate limbs.

We review embryological as well as molecular evidence that emphasizes the idea that both the regenerate and the developing vertebrate limb bud utilize a similar set of signals that regulate pattern formation. Evidence is presented to implicate the Hox-7.1 gene in the developmental regulation of growth, differentiation, and positional assignment during limb outgrowth and the proposal is made that the expression of this gene governs the cellular activities within the progress zone during limb outgrowth. Finally, we review the limited information known about the regenerative capabilities of limb buds in organisms that cannot regenerate as adults. We content that a solution to the problem of regenerative failure among higher vertebrates will come progressively through a stepwise analysis of impaired regeneration associated with increasing developmental age.

Animals↗
PubMed1992

Position-specific growth of mouse limb bud cells in vitro.

The relationship between cellular position and growth control has been studied in cultures of dissociated fragments of mouse limb bud cells. Using cells derived from various positions along the anterior-posterior axis of the limb bud we have developed culture conditions that optimize growth of positionally isolated cells. Under these conditions limb bud cells display an inherent, position-specific growth response; proliferation of cells derived from anterior and central regions of the limb is enhanced over that of posterior derived cells. Thus, within the total population of limb bud cells the in vitro growth of posterior cells is unique and correlates with the positional activity associated with the zone of polarizing activity. Anterior and posterior cells were cocultured to determine whether interactions between these two groups of positionally distinct cells lead to the stimulation of growth that has been observed in vivo. We observe a slight but consistent position-dependent stimulation of growth that is indicative of a mitogenic signal passing between these positionally disparate cells. Similarities between position-related growth dynamics in vivo and in vitro suggest that positional interactions that are important for limb formation can occur between dissociated cells cultured under standard conditions.

Animals↗
PubMed1991

Conversion by retinoic acid of anterior cells into ZPA cells in the chick wing bud.

In recent years there has been considerable interest in the role of retinoic acid (RA) in vertebrate-limb pattern formation. When RA is applied to the anterior of the chick wing bud, a mirror-image duplication of the limb pattern develops that is identical to the pattern resulting from grafts of posterior tissue (zone of polarizing activity, or ZPA). It has been proposed that position along the anterior-posterior axis in the chick limb is specified by a gradient of a diffusible factor produced by the ZPA. The ZPA-mimicking action of RA has led to the hypothesis that exogenously applied RA acts by providing graded spatial information across the anterior-posterior limb axis. An alternative interpretation is that RA changes anterior cells into ZPA cells, which in turn provide the actual pattern-duplicating stimulus; there is already some preliminary evidence that this occurs. A hybrid interpretation has also been suggested whereby ZPA cells are formed in response to RA exposure and then begin to release retinoids that act as graded spatial cues. We have used a functional assay to test anterior chick wing-bud cells for ZPA activity after exposure to RA. The results of our studies indicate that the action of RA is to change anterior cells into ZPA cells. Further, our results indicate that it is unlikely that RA-treated anterior cells then begin producing RA in such a way as to provide a graded positional signal.

Animals↗
PubMed1991

Mapping the early development of projections from the entorhinal cortex in the embryonic mouse using prenatal surgery techniques.

The purpose of this work was to study the development of specific projections from the postero-lateral cortex during the third trimester of gestation in the mouse. To do this, we labeled undifferentiated lateral cortex with the fluorescent carbocyanine dye, Dil, in the embryonic day (E) 16 mouse embryo using exo utero surgical techniques (Muneoka, Wanek, and Bryant, 1986). Embryos were allowed to develop to term (postnatal day 0, P0) at which time the fiber patterns emanating from the marked regions were studied. Dye placement in the undifferentiated postero-ventral cortex produced labeled fibers in the hippocampal formation. A robust projection of the angular bundle into the CA1 region of the hippocampus was heavily labeled. In addition, in some animals, cortical tracts, such as the anterior commissure, corpus callosum, and a corticotectal tract, were labeled. These tracts have been described previously as scaffolding pathways in the fetal cat (McConnell, Ghosh, and Shatz, 1989), and other vertebrates (Wilson, Ross, Parrett, and Easter, 1990). Dye placement in adjacent, more anterior or dorsal areas showed strong labeling in cortical structures but no labeling in the hippocampal formation. These data indicate that, by birth, the temporal cortex is subdivided along the rostro-caudal axis as entorhinal cortex and perirhinal cortex, and along the dorso-ventral axis, as entorhinal cortex and neocortex. Also, these earliest connections are similar to adult connections in their specificity of target area selection. Therefore, these early, yet specific, connections may play a role int he formation of future connections during postnatal development.

Animals↗
PubMed1989

A staging system for mouse limb development.

A series of 15 stages of development for the mouse limb bud have been defined, spanning the time from the first appearance of the limb bud to the completion of limb outgrowth. The stages are based on changes in the morphology of the limb in living preparations. The development and regression of the apical ectodermal ridge (AER) as well as the development of the skeletal structures are also described. This staging system has been developed in response to the need to standardize in situ experimental analyses of the mouse limb bud. Comparable stages of the commonly used chick wing and mouse whole embryo systems are presented.

Animals↗
PubMed1989

Mammalian limb bud development: in situ fate maps of early hindlimb buds.

Fate maps of the developing mouse hindlimb bud have been constructed for the first time using exo utero surgical techniques and carbon particle injections. Such fate maps demonstrate that the limb develops in a proximal to distal manner as a result of distal expansion. The anterior-posterior extent of the limb bud develops asymmetrically with the posterior half giving rise to slightly more of the digit pattern (digits 3-5) than the anterior half (digits 1 and 2). We found no evidence for the occurrence of extensive cellular rearrangements during limb development, and the free limb bud appears to give rise to only zeugo- and autopodial elements with the stylopod arising in the body wall proximal to the bud. These results are consistent with our current understanding of limb development in lower vertebrates and also provide detailed information that will be useful for future limb studies in mammals.

Animals↗
PubMed1989

Evidence for regulation following amputation and tissue grafting in the developing mouse limb.

Procedures are now available to experimentally manipulate postimplantation mouse embryos in situ and allow development to continue into postnatal life (Muneoka, K., N. Wanek, and S.V. Bryant (1986) J. Exp. Zool., 239:289-293). We have investigated the ability of the well-formed hindlimb bud to regulate following two experimental operations: amputation and wedge grafts designed to confront anterior and posterior cells. After comparing the resultant limbs with the fate maps of the relevant stages, we conclude that the developing hindlimb bud at stage 7/8 (equivalent to stage 27/28 of the chick) is capable of partial regeneration of the peripheral digits following amputation and capable of supernumerary digit tip formation after grafting of a wedge of anterior tissue to a posterior position.

Animals↗
PubMed1987

Intercalation and the cellular origin of supernumerary limbs in Xenopus.

The hypothesis that a specialized polarizing zone controls the pattern of the anterior-posterior axis during limb development in Xenopus has been tested by analysing the cellular contribution to supernumerary limbs. Supernumerary limbs were generated by grafting hindlimb buds contralaterally between X. borealis and X. laevis to appose anterior and posterior limb tissues. Cells derived from these two species of Xenopus are readily identified by staining with quinacrine. The analysis of cellular contribution showed that supernumerary limbs consist of approximately half anterior-derived (57%) and half posterior-derived (43%) cells. These data are not consistent with the polarizing zone theory but are consistent with the hypothesis that both supernumerary limbs and normally developing limbs arise from intercalary interactions between limb bud cells with different positional values.

Animals↗
PubMed1986

Mouse embryos develop normally exo utero.

Surgical procedures are described which permit direct experimental access to postimplantation mouse embryos. These procedures arose from our finding that development proceeds normally to term after embryos have been released from the confines of the uterus, while remaining attached to it via the placenta. Embryos continue to develop exo utero, within the abdominal cavity of the mother, and are capable of surviving a variety of different surgical manipulations. This finding opens the way for experimental analyses of mouse development in vivo.

Animals↗
PubMed1986

Intrinsic control of regenerative loss in Xenopus laevis limbs.

The regenerative capability of Xenopus laevis hindlimbs was studied at different developmental stages. Three types of surgical deletion of the autopod were performed: simple amputations, 3-digit wedge-shaped deletions and 1-digit wedge-shaped deletions. The frequency of regenerative response and the digital patterns of the regenerates were analyzed. Regenerative capacity declines with developmental stage for all three types of deletions. For simple amputations this decline occurs in an orderly manner with the failure of anterior digits to regenerate at progressively later stages. A comparison between 1-digit and 3-digit deletions shows that for all stages of development, 3-digit deletions regenerate better than one-digit deletions. These data indicate that the amount of tissue removed is directly related to whether a regenerative response is observed. At any given stage, larger deletions are more likely to regenerate than smaller ones. These results are discussed with regard to the mechanisms by which growth during regeneration is controlled.

Animals↗
PubMed1986

The migration of dermal cells during blastema formation in axolotls.

Using the diploid/triploid cell marker in the axolotl (Ambystoma mexicanum) we have examined the movement of cells from the dermis into the early limb blastema. Cells of dermal origin begin to migrate beneath the wound epithelium at about 5 days postamputation, and by 10 days they are widely distributed across the amputation surface. By 15 days, a dense accumulation of blastema cells is present beneath the apical cap, and these cells are preferentially oriented in a circumferential direction. These results are discussed in relation to previous studies showing that the progeny of dermal cells become widely distributed during regeneration, and that cells of dermal origin are a major source of blastema cells. The results are also discussed in relation to ideas about how growth and patterning of the new appendage occur.

Ambystoma mexicanum↗
PubMed1986

Cellular contribution from dermis and cartilage to the regenerating limb blastema in axolotls.

Using the triploid/diploid cell marker in the axolotl, Ambystoma mexicanum, we have analyzed the extent to which cells derived from the dermis and the skeleton contribute to the regenerating limb blastema. We found that dermal cells contribute 43% of the blastemal cell population whereas cells derived from skeletal tissue contribute only 2%. When compared to the availability of cells at the plane of amputation, dermal cells overcontribute by greater than twofold whereas skeletal cells undercontribute by several-fold. These data correlate with the effects that these two tissues have on the formation of the limb pattern during regeneration; dermis has a dramatic influence on pattern and skeletal tissue has virtually no effect. It is suggested that the fibroblasts present in the dermis and in other parts of the limb form virtually all of the mesodermal tissues in the regenerate with the exception of the muscle.

Ambystoma mexicanum↗
PubMed1986

Pattern discontinuity, polarity and directional intercalation in axolotl limbs.

Axolotl limb stumps with dorsal-ventral confrontations between digits 2 and 3 but with a normal anterior-posterior pattern were created by grafting between contralateral limbs. Graft and host differed in ploidy to permit a determination of the origin of cells in the regenerated limb. After regeneration, limbs were analysed for skeletal and muscle patterns and for the distribution of marked cells in the regenerate. Regenerated limbs showed varying degrees of abnormality in their dorsal-ventral organization. Following regeneration, the original dorsal-ventral discontinuities were in some cases maintained and in others resolved. The maintenance or resolution of pattern discontinuities occurred in a position-dependent manner. Cell marker analysis indicates a relationship between the resolution of discontinuities and the extent to which cells become displaced across the original graft-host interface. These data lend support to the suggestion that circumferential intercalation is directionally biased.

Ambystoma↗