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D L Stocum

Publications and source records attributed to D L Stocum.

At least 37 records · Page 2Linked to original sources
PubMed1988

Retinoic acid coordinately proximalizes regenerate pattern and blastema differential affinity in axolotl limbs.

An assay that detects position-related differences in affinity of axolotl regeneration blastema cells in vivo was used to test whether retinoic acid, which proximalizes regenerate pattern, simultaneously proximalizes blastema cell affinity. The assay involved autografting or homografting late bud forelimb blastomas derived from the wrist, elbow or midupper arm levels to the dorsal surface of the blastema-stump junction of an ipsilateral, medium-bud-stage hindlimb regenerating from the midthigh level. The grafted blastemas consistently displaced to their corresponding levels on the proximodistal axis of the host regenerate, indicating the existence of level-specific differences in blastema cell affinity. Retinoic acid proximalized the pattern of donor forelimb regenerates to the level of the girdle and abolished their displacement behaviour on untreated host hindlimbs. Conversely, untreated forelimb donor blastemas displaced distally to their corresponding levels on host ankle regenerates, that had been proximalized to the level of the girdle by retinoic acid. These results indicate that positional memory in regenerating limbs is directly related to blastema cell affinity, and that very similar or identical sets of level-specific affinity properties are shared by forelimb and hindlimb cells.

Ambystoma↗
PubMed1988

Retinoic acid proximalizes level-specific properties responsible for intercalary regeneration in axolotl limbs.

The objective of this study was to determine whether retinoic acid (RA) coordinately proximalizes positional memory and the cellular recognition system that detects pattern discontinuity in regenerating amphibian limbs. The strategy was to test the capacity of RA-treated blastemas to evoke intercalary regeneration when grafted to an amputation level proximal to their level of origin. Control wrist and ankle, or elbow and knee blastemas treated with the retinoid solvent, dimethylsulphoxide, evoked intercalary regeneration as effectively as untreated blastemas, when grafted to the midstylopodial amputation surface of host limbs. RA-treated wrist and ankle or elbow and knee blastemas were proximalized and formed complete limbs that were at an angle to, or continuous with, the midstylopodium of the host limb. No intercalary regeneration, from either graft or host, was observed in these cases. The results indicate that the cellular mechanism that recognizes disparities between non-neighbouring cells and initiates intercalary regeneration is coordinately proximalized with positional memory. Thus the recognition mechanism and positional memory are directly related. Intercalary regeneration and corrective displacement (affinophoresis), both of which restore a pattern of normal cell neighbours by different means in regenerating axolotl limbs, appear to use the same mechanism to recognize pattern discontinuity.

Ambystoma↗
PubMed1987

Use of retinoids to analyze the cellular basis of positional memory in regenerating amphibian limbs.

Cells of the amphibian limb regeneration blastema inherit memories of their level of origin (positional memory) along the limb axes. These memories serve as boundaries of what is to be regenerated, thus preventing regeneration of any but the missing structures. Because of its importance in determining the boundaries of regenerate pattern, it is essential to understand the cellular and molecular basis of positional memory. One approach to this problem is to look for position-related differences in a cell or molecular property along a limb axis and then show, using an agent that modifies regenerate pattern, that the cell or molecular property and the pattern are coordinately modified. We have done this using retinoic acid (RA) as a pattern-modifying agent and an in vivo assay that detects position-related differences in a cell recognition-affinity property along the proximodistal (PD) axis of the regenerating axolotl limb. RA proximalizes positional memory in the PD axis, posteriorizes it in the anteroposterior axis, and ventralizes it in the dorsoventral axis. The level-specific PD cell recognition-affinity property is proximalized by RA, indicating that this property and positional memory are causally related. The effects of RA on positional memory may be mediated through a cellular RA-binding protein (CRABP), since the concentration of unbound (apo) CRABP molecules is highest during early stages of regeneration when the proximalizing effects of RA are greatest.

Amphibians↗
PubMed1986

Retinoic acid modifies positional memory in the anteroposterior axis of regenerating axolotl limbs.

The effects of retinoic acid (RA) on anteroposterior (AP) positional memory of regenerating axolotl limbs were tested after removing the anterior or posterior half from the zeugopodium (lower arm or leg). RA (150 micrograms/g body wt) was injected into groups of animals bearing the following types of limbs: (1) anterior and posterior half zeugopodia grafted to the eyesocket and amputated distally 7 days later; (2) unamputated anterior and posterior half zeugopodia in situ; (3) double anterior and double posterior half zeugopodia amputated distally 7 days after their construction; (4) sham-operated zeugopodia amputated distally 7 days after operation. Controls consisted of these four groups injected with the retinoid solvent, dimethyl sulfoxide, or not injected. Control half zeugopodia grafted to the eyesocket regenerated no more than one or two digits. Control unamputated half zeugopodia in situ underwent partial or complete regeneration of the missing half from the proximal and midline wound surfaces exposed during construction of the half zeugopodia. Control double anterior and posterior zeugopodia both regenerated symmetrical, hypomorphic regenerates with 1-3 digits in the double anteriors and 1-6 digits in the double posteriors. Sham-operated controls regenerated normally. Regenerating anterior and posterior halves responded differently to RA. RA-treated anterior half zeugopodia in the eyesocket, and anterior half stumps adjacent to the unamputated posterior half zeugopodia in situ both produced regenerates that duplicated stump structures in the proximodistal axis and formed a complete and normal AP pattern. RA-treated double anterior zeugopodia regenerated proximodistal-duplicated pairs of mirror-imaged limbs, each with a complete and normal AP pattern. In contrast, half posterior zeugopodia in the eyesocket, the posterior half stumps of unamputated half anterior zeugopodia in situ, and double posterior zeugopodia all failed to regenerate. These results suggest that RA modifies positional memory in only one direction in the AP axis, posterior.

Ambystoma↗
PubMed1984

Retinoic-acid-induced pattern completion in regenerating double anterior limbs of urodeles.

The effects of retinoic acid on the regeneration of double anterior lower arms in the adult newt, Notophthalmus viridescens, were investigated. Normally, double anterior lower arms regenerate a hypomorphic, symmetrical pattern of structures, which are distally complete; and double anterior upper arms regenerate a hypomorphic, symmetrical but distally incomplete pattern of structures. In limbs with a normal anteroposterior axis, the major effect of retinoic acid is to alter the proximodistal (PD) positional value of cells local at the amputation level to a much more proximal value, thereby creating duplications in the regenerate of structures proximal to the amputation plane (Thoms and Stocum, '84). Therefore, we predicted that double anterior lower arms treated with retinoic acid would regenerate like double anterior upper arms. However, in a substantial number of cases, each half of these double anterior lower arms regenerated a limb that was complete in the anteroposterior (AP) axis, with asymmetry corresponding to the half of origin. In addition, these regenerates were serially duplicated in the PD axis. These results indicate that retinoic acid can posteriorize the positional value of midline cells, leading to restoration of normal AP pattern, when the set of posterior-half positional values is removed from the cross section of the limb.

Amputation, Surgical↗
PubMed1984

Retinoic acid-induced pattern duplication in regenerating urodele limbs.

The effects of varying doses of retinoic acid on forelimb regeneration in larval Ambystoma mexicanum amputated through the wrist joint and in adult Notophthalmus viridescens amputated through the basal carpals were compared. In both species, the major effect of retinoic acid was to cause the proximodistal duplication, in the regenerate, of stump segments proximal to the amputation plane. Transverse axial duplications (anteroposterior and dorsoventral) occurred in a smaller percentage of cases; these consisted of cartilage spurs in axolotls, and extra digits in newts. The frequency and magnitude of the proximodistal and (in the newt) transverse duplications were dose dependent, and the regenerating limbs were maximally sensitive to the retinoid during the period of dedifferentiation and accumulation of blastema cells. The effect of retinoic acid is exerted on cells local to the amputation surface, as shown by the fact that retinoic acid caused the proximodistal duplication of stump segments in regenerates derived from amputated distal lower arm segments grafted to the eyesocket .

Ambystoma↗
PubMed1984

The urodele limb regeneration blastema. Determination and organization of the morphogenetic field.

The idea that the undifferentiated limb regeneration blastema of urodele amphibians is an undetermined and pluripotent structure is examined. A detailed review of the literature shows that this notion has no basis in fact. The data show that the morphogenetic potency of the blastema is restricted to its prospective significance and that this potency can be fully expressed when the blastema is transplanted either to a neutral location or to a regenerating organ of another type. Within this morphogenetic constraint, however, blastema cells have a histogenetic potency that is, at least in some cases, greater than their limb cell phenotype of origin. The morphogenetic responses of the regeneration field to discontinuities suggest that its autonomous determining relationships are based on the inheritance, from parent limb cells, of a graded set of mesodermal positional values specifying the pattern of the amputation plane, and a single epidermal external boundary value. The dividing mesenchymal cells of the blastema change positional value to erase any discontinuity between themselves and the epidermis, and the epidermis acts as a stop signal to inform the mesenchyme when the regenerate boundary has been reached. In vitro experiments suggest that changes in mesenchymal positional value in response to discontinuity can be interpreted in terms of gradients of cell-cell adhesivity, and they focus attention on the importance of molecular studies of blastema cell surfaces for our future understanding of regeneration and morphogenesis in general.

Animals↗
PubMed1982

Control of pattern formation in urodele limb ontogeny: a review and a hypothesis.

From a review of the literature, the hypothesis is advanced that the forelimb region of the urodele embryo acquires its transverse axial polarity and pattern by the action of posterior and dorsal polarizing regions. The anterior-posterior and dorsal-ventral axes are determined simultaneously and their determination is a prerequisite for proximal-distal outgrowth. Outgrowth of the limb bud is accompanied by the generation, between proximal and distal boundaries, of a set of positional values specifying proximal-distal axial polarity and pattern. The proximal boundary is the initial positional value carried by the cells of the limb area. The distal boundary is imposed upon the outermost layer of limb disc cells by the overlying ectoderm.

Ambystoma↗
PubMed1982

Determination of axial polarity in the urodele limb regeneration blastema.

The state of determination of the anterior-posterior, dorsal-ventral and proximal-distal axes of the undifferentiated limb regeneration blastema was evaluated by heterografting and autografting experiments in which these axes were reversed with respect to the limb stump. Species-specific size differences in skeletal elements were used as markers to trace the origin of regenerate tissues in the heterografting experiments, and differences in the skeletal patterns of hindlimbs and forelimbs were used as markers in the autografting experiments. The resulting primary regenerates fell into two categories, those composed wholly or partly of donor tissues, and those composed entirely of host tissues. Regenerate structures formed from donor tissues always maintained the handedness of origin, while regenerates formed from host tissues always displayed host-side handedness. These results demonstrate that the axes of the blastema are determined from the start of regeneration, and that previous claims of axial lability in reversal experiments are based on an illusion created by resorption of graft tissue, accompanied by regeneration from the host. Reversal of the transverse axes resulted in the formation of supernumerary limbs. Analysis of heterograft cases in which the anterior-posterior axis was reversed showed that 50% of the supernumeraries were constructed partly of donor blastema tissue whose axial polarity was reversed with respect to the adjacent primary regenerate. The vast majority of the primary regenerates in these cases possessed the normal number of digits. It is thus likely that the reprogrammed donor blastema cells used to construct the supernumeraries are derived by division of a thin band of cells at the edge of the graft adjacent to the supernumerary.

Ambystoma↗
PubMed1979

Stages of forelimb regeneration in Ambystoma maculatum.

A series of normal stages describing the regeneration of larval A. maculatum limbs after amputation through the upper arm or wrist is described. Nine discrete stages were recognized, based on external morphological and associated histological features. These stages are Initial Dedifferentiation (ID), Early Bud (EB), Medium Bud (MB), Late Bud (LB), Early Redifferentiation (ER), Notch (N), 2-Fingerbud (2-FB), 3-Fingerbud (3-FB) and 4-Fingerbud (4-FB). Similarities and differences between this and other staging systems for urodele limb regeneration are discussed. The absence of osteoclasts was a striking feature during dedifferentiation of the wrist, in contrast to their presence in large numbers during dedifferentiation of the upper arm.

Ambystoma↗
PubMed1979

Development of the tibiotarsus in the chick embryo: biosynthetic activities of histologically distinct regions.

The dynamics of the histological changes which occur in the distal half of the tibial portion of the embryonic chick tibiotarsus from day 8 to day 18 of incubation are correlated with the capacities of histologically distinct zones to incorporate isotopic precursors into mucopolysaccharides and collagen. At the distal end of the tibia, which abuts the suture line formed by the fusion of the two tarsals with the tibia, there persists throughout embryonic development a narrow band of small, round or oval, rapidly dividing chondrocytes which synthesize chondroitin SO4 and collagen at low rates. Just proximal to this zone is a broader zone of flattened, disc-shaped chondrocytes which divide more slowly and are extremely active in chondroitin SO4 and collagen synthesis. Proximal to the zone of flattened chondrocytes is a zone of non-dividing, hypertrophied chondrocytes which are large and round and increase continually in size going from the distal to the proximal end of the zone. The biosynthetic activities of the cells in this zone fall sharply with their distance from the zone of flattened chondrocytes. Finally, there is a fourth zone, the marrow cavity, formed by a proximal to distal disintegration of the hypertrophied chondrocytes, starting at mid-diaphysis. The marrow cavity is surrounded by a shell of periosteal and intra-membranous bone which extends to the distal end of the zone of hypertrophied chondrocytes. Our data suggest that as the tibiotarsus grows in length the small, round rapidly dividing cells of the tibia differentiate first to flattened chondrocytes which synthesize matrix at a high rate and ultimately to low activity, hypertrophying chondrocytes. This sequence proceeds in a linear fashion.

Animals↗
PubMed1978

Regeneration of symmetrical hindlimbs in larval salamanders.

The complete circle rule of the polar coordinate model of pattern regulation was tested for regenerating hindlimbs of Ambystoma larvae. The hindlimbs were made symmetrical in the circumference of either the thigh or the shank, and their ability to regenerate from both levels was observed. Thighs composed of two anterior halves failed to regenerate, whereas thighs composed of two posterior halves often regenerated distally complete, correspondingly symmetrical limbs; shanks composed of either two anterior or two posterior halves regenerated distally complete, correspondingly symmetrical limbs. These results are in contrast to what is predicted by the complete circle rule and suggest a modification of this rule.

Ambystoma↗