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L C Javois

Publications and source records attributed to L C Javois.

At least 19 recordsLinked to original sources
PubMed1999

Interactions between the foot and the head patterning systems in Hydra vulgaris.

The Cnidarian, hydra, is an appealing model system for studying the basic processes underlying pattern formation. Classical studies have elucidated much basic information regarding the role of development gradients, and theoretical models have been quite successful at describing experimental results. However, most experiments and computer simulations have dealt with isolated patterning events such as the dynamics of head regeneration. More global events such as interactions among the head, bud, and foot patterning systems have not been extensively addressed. The characterization of monoclonal antibodies with position-specific labeling patterns and the recent cloning and characterization of genes expressed in position-specific manners now provide the tools for investigating global interactions between patterning systems. In particular, changes in the axial positional value gradient may be monitored in response to experimental perturbation. Rather than studying isolated patterning events, this approach allows us to study patterning over the entire animal. The studies reported here focus on interactions between the foot and the head patterning systems in Hydra vulgaris following induction of a foot in close proximity to a head, axial grafting of a foot closer to the head, or doubling the amount of basal tissue by lateral grafting of an additional peduncle-foot onto host animals. Resulting positional value changes as monitored by antigen (TS19) and gene (ks1 and CnNK-2) expression were assessed in the foot, head, and intervening tissue. The results of the experiments indicate that positional values changed rapidly, in a matter of hours, and that there were reciprocal interactions between the foot and the head patterning systems. Theoretical interpretations of the results in the form of computer simulations based on the reaction-diffusion model are presented and predict many, but not all, of the experimental observations. Since the lateral grafting experiment cannot, at present, be simulated, it is discussed in light of what has been learned from the axial grafting experiments and their simulations.

Animals↗
PubMed1999

Interactions between the foot and bud patterning systems in Hydra vulgaris.

In the freshwater coelenterate, hydra, asexual reproduction via budding occurs at the base of the gastric region about two-thirds of the distance from the head to the foot. Developmental gradients of head and foot activation and inhibition originating from these organizing centers have long been assumed to control budding in hydra. Much has been learned over the years about these developmental gradients and axial pattern formation, and in particular, the inhibitory influence of the head on budding is well documented. However, understanding of the role of the foot and potential interactions between the foot, bud, and head patterning systems is lacking. The purpose of this study was to investigate the role of the foot in the initiation of new axis formation during budding by manipulating the foot and monitoring effects on the onset of first bud evagination and the time necessary to reach the 50% budding point. Several experimental situations were examined: the lower peduncle and foot (PF) were injured or removed, a second PF was laterally grafted onto animals either basally (below the budding zone) or apically (above the budding zone), or both the head and PF were removed simultaneously. When the PF was injured or removed, the onset of first bud evagination was delayed and/or the time until the 50% budding point was reached was longer. The effects were more pronounced when the manipulation was performed closer to the anticipated onset of budding. When PF tissue was doubled, precocious bud evagination was induced, regardless of graft location. Removal of the PF at the same time as decapitation reduced the inductive effect of decapitation on bud evagination. These results are discussed in light of potential signals from the foot or interactions between the foot and head patterning systems that might influence bud axis initiation.

Animal Structures↗
PubMed1996

The dynamics of head activation changes during proportioning in Hydra oligactis with altered head-body ratios.

Normal hydra head-body proportions were altered by axially grafting a second head in place of the lower body column. The resulting animals had double the head tissue and one-quarter the normal body column. Changes in the head activation potential of tissue subjacent to both heads were monitored by assaying the ability of these animals to regenerate heads. The host head, the grafted head, or both heads were removed at varying times following graft construction and the animals were scored for head regeneration and/or the ability to express a head-specific antigen recognized by monoclonal antibody, CP8. In the presence of the grafted head, tissue subjacent to the host head lost the ability to regenerate a head or express the head-specific antigen over a 48-hr period. In the presence of the host head, tissue subjacent to the grafted head regenerated heads at a very low frequency and lost the ability to express the head-specific antigen over the same 48-hr period. Following simultaneous removal of both heads, animals initially regenerated both heads for the first 48 hr after graft construction. Then, both head regeneration and expression of the head-specific antigen declined gradually over the next 3 days, though not to the very low levels observed when one head remained. These data, especially the loss of CP8 labeling, support the hypothesis that loss of regeneration ability was due to a loss of head activation potential in tissues subjacent to the heads. We propose that this reflected the attempt of grafted animals to compensate for the altered head-body proportions through reproportioning. In keeping with this hypothesis, feeding the grafted animals to stimulate growth of body column tissue and a shift toward more normal head-body proportions prior to decapitation resulted in animals which were capable of regenerating heads when decapitated. Several interpretations of the results based on the Gierer-Meinhardt reaction-diffusion model of pattern formation are discussed.

Animals↗
PubMed1993

Patterning of heads and feet during regeneration of Hydra oligactis aggregates.

The de novo appearance of head organizing centers was monitored during regeneration of aggregates made from dissociated body column tissue of Hydra oligactis using the head-specific monoclonal antibody CP8 (L. C. Javois, R. D. Wood, and H. R. Bode, 1986, Dev. Biol. 117, 607-618). Newly differentiated CP8+ head organizing centers were present by 24 hr of regeneration and were restricted to one half of the aggregate. Depending on subsequent patterning events these CP8+ cells went on to develop into heads or body tentacles, or remained as CP8+ patches. A comparison of the number of initial CP8+ patches with the final number of CP8+ structures indicated that this number was fairly constant, suggesting that a limited number of head organizing centers were established early during regeneration. Examination of fully differentiated head structures revealed that they were segregated to one end of the aggregate with feet segregated away from the CP8+ structures. Increasing the surface area of the aggregates resulted in a more random initial spacing of CP8+ head organizing centers and ultimately a more random distribution of differentiated heads and feet. However, polarized polyps with one head and foot detached and the remaining structures were then segregated. The ability of existing pattern formation models to describe interactions between head- and foot-forming systems resulting in the observed spacing of heads and feet is discussed.

Animals↗
PubMed1992

Nerve ring of the hypostome in hydra. I. Its structure, development, and maintenance.

The anatomy and developmental dynamics of the nerve ring in the hypostome of Hydra oligactis were examined immunocytochemically with an antiserum against a neuropeptide and with neuron-specific monoclonal antibodies. The nerve ring is unique in the mesh-like nerve net of hydra. It is a distinct neuronal complex consisting of a thick nerve bundle running circumferentially at the border between the hypostome and tentacle zone. Immunostaining showed that the nerve ring was heterogeneous and contained at least four different subsets of neurons. During head regeneration and budding, the nerve ring appeared only after the nerve net of ganglion and sensory cells had formed. Every epithelial cell is continuously displaced with neurons toward either head or foot in an adult hydra. However, the ectoderm in the immediate vicinity of, and including, the nerve ring constitutes a stationary zone that is not displaced. Tissue immediately above this zone is displaced toward the tip of the hypostome, while tissue below is displaced along the tentacles. Correspondingly, the production of new neurons in the ring as measured by their differentiation kinetics is much slower than in surrounding areas. Thus, the nerve ring is static and stable in contrast to the dynamic features of the nerve net of hydra.

Animals↗
PubMed1991

Simultaneous effects of head activator on the dynamics of apical and basal regeneration in Hydra vulgaris (formerly Hydra attenuata).

Previous studies have demonstrated that head activator (HA), an 11 amino acid peptide, stimulates head-specific differentiation processes in hydra. Additionally, HA enhances the differentiation of interstitial cells into nerve cells. This study investigated the effects of exogenous synthetic HA on the dynamics of both apical and basal regeneration in a piece of tissue excised from the body column of treated animals which comprised one-eighth of the original animal. The dynamics of apical and basal regeneration were monitored using the monoclonal antibody TS19. This antibody binds to apical and basal ectodermal tissue very early in the process of regeneration, before morphological structures are evident. Labeling is ultimately localized to the tentacles of the head and a ring above the basal disc. Thus, TS19 is a useful tool for analyzing the dynamics of both apical and basal patterning processes in the same regenerate simultaneously. Quantification of TS19 positive areas on regenerates over a time course of 72 hr revealed that HA treatment accelerated and amplified the dynamics of both apical and basal TS19 labeling. The specific basal effect was novel and was demonstrated to occur in the absence of a determined head independently of new nerve cell differentiation. It is proposed that the basal effect was the result of growth factor-like activity of HA.

Animals↗
PubMed1990

Patterning of the head in hydra as visualized by a monoclonal antibody: III. The dynamics of head regeneration.

The dynamics of the early patterning processes leading to the regeneration of a head in tissue excised from the body column of Hydra oligactis were examined by using a monoclonal antibody, CP8. This antibody displays position-specific binding, labeling the head ectodermal epithelial cells. During regeneration of a head, antibody labeling is present well before morphological signs of the head, at a time correlated with the determination of the tissue (Javois et al., Dev. Biol., 117:607-618, '86). By quantifying antibody labeling during regeneration of three different pieces of tissue excised from the body column, it was found that the dynamics of the early patterning processes as visualized by CP8 labeling varied. The pattern of labeling observed as well as the spread of labeled tissue suggested that the amount and geometry of apical tissue in the regenerate played a critical role in the patterning processes. Contrary to the labeling pattern observed in heads which formed during bud development or which regenerated following decapitation (Javois et al., '86), not all the CP8+ tissue was confined to the head structures in these regenerates. Several alternative explanations for this surprising result are presented. The usefulness of these data in refining pattern formation models by more explicitly constraining their parameters is discussed.

Animals↗
PubMed1990

Effect of head activator on proliferation, head-specific determination and differentiation of epithelial cells in hydra.

In hydra the differentiation of head-specific ectodermal epithelial cells from multipotent stem cells is a multistep process in which cell cycle progression is regulated at three restriction points. Head activator acts as a positive signal at these restriction points. At the G2/mitosis boundary of epithelial stem cells head activator functions as a mitogen, being necessary for cell division. Subsequently, in or before S phase, head activator acts as determinant to ensure commitment of epithelial cells to head-specific determination. This effect of head activator requires hundredfold-higher concentrations, and may also require longer incubation times, than for cell proliferation. Epithelial cells thus committed to head-specific differentiation become arrested in G2 as a third and last restriction point in the cell cycle. They require disinhibition by decapitation and probably the presence of head activator for final differentiation, which then occurs in G2.

Animals↗
PubMed1988

Patterning of the head in hydra as visualized by a monoclonal antibody, II. The initiation and localization of head structures in regenerating pieces of tissue.

The body column of hydra is polarized such that a new head will regenerate from the apical end when both extremities are removed. This is due to a graded property of the tissue termed the head activation gradient. The aim of the experiments presented here was to determine what events connect a two-dimensional segment of the activation gradient in an isolated piece of tissue with the formation of a head structure at a particular location. To this end, tissue pieces with three different shapes were excised and analyzed during and after regeneration. The most apical tissue of each piece was labeled with the DNA-intercalating dye, DAPI, and the area where developmental changes were occurring was monitored using the monoclonal antibody CP8 (Javois et al., 1986). First, it was shown that polarity of regeneration was maintained regardless of the fraction of body length included in the excised pieces. Second, while head structures usually formed from the original apical tissue, they could be located anywhere in the regenerate. This was an effect of the healing process which shaped the apical edge differently in different pieces. Third, early CP8 binding occurred in similarly shaped areas suggesting that patterning events were initiated in a contiguous manner wherever apical tissue was located. And finally, not all of the CP8-marked tissue successfully formed structures. Apparently some regions were favored to continue the patterning process, and these in turn extinguished the process in neighboring regions.

Animals↗
PubMed1986

Patterning of the head in hydra as visualized by a monoclonal antibody. I. Budding and regeneration.

A monoclonal antibody, CP8, has been isolated which displays a position-specific binding pattern to epithelial cells of Hydra oligactis. Antibody binding is restricted to the head of adult animals. When a new head develops during the budding process, CP8 binding is present in the area which will form the head well before morphological signs of it. Similarly, following decapitation as a new head regenerates, CP8 label appears covering a domed area at the apical end of the regenerate before tentacles evaginate delineating the head. When bud development or regeneration is complete, CP8 label is restricted to the new head. Experiments indicate the appearance of CP8 label during the formation of a head correlates closely with the patterning events which result in the determination of the tissue to form a head. The usefulness of CP8 as a diagnostic tool for exploring the dynamics of head pattern formation in hydra is discussed.

Animals↗