Posterior hoxa genes expression during zebrafish bony fin ray development and regeneration suggests their involvement in scleroblast differentiation.

Expression of two zebrafish developmental posterior hoxa genes, hoxa11b and hoxa13b, was studied by in situ hybridization during pectoral and caudal fin development and regeneration. Expression was restricted to cells of the bony rays region. During fin development, molecular cytological analysis revealed that a subpopulation of mesenchymal cells expressed these two hoxa genes during their early differentiation in the subapical region of the developing ray.

Comparison of even-skipped related gene expression pattern in vertebrates shows an association between expression domain loss and modification of selective constraints on sequences.

The even-skipped related genes (evx) encode homeodomain-containing transcription factors that play key roles in body patterning and neurogenesis in a wide array of Eumetazoa species. It is thought that the genome of the last common ancestor of Chordata contained a unique evx gene linked to a unique ancestral Hox complex. During subsequent evolution, two rounds of whole genome duplication followed by individual gene losses gave rise to three paralogs: evx1, evx2, and eve1.

Old questions, new tools, and some answers to the mystery of fin regeneration.

Pluridisciplinary approaches led to the notion that fin regeneration is an intricate phenomenon involving epithelial-mesenchymal and reciprocal exchanges throughout the process as well as interactions between ray and interray tissue. The establishment of a blastema after fin amputation is the first event leading to the reconstruction of the missing part of the fin. Here, we review our knowledge on the origin of the blastema, its formation and growth, and of the mechanisms that control differentiation and patterning of the regenerate.

Zebrafish keratin 8 is expressed at high levels in the epidermis of regenerating caudal fin.

Cytokeratins are structural proteins of the intermediate filament family and are mainly expressed in epithelial cells. In several vertebrates it has been shown that keratin 8 is expressed in simple epithelial tissues, some non-epithelial tissue and in hyper-proliferative tissues during development and tumor transformation. We previously cloned and characterised the zebrafish (Danio rerio) homologous cytokeratin 8 cDNA (zfk8) which was described as an epidermal marker during zebrafish development.

evx1 transcription in bony fin rays segment boundaries leads to a reiterated pattern during zebrafish fin development and regeneration.

The dermoskeleton of zebrafish fins is composed of actinotrichia and segmented bony rays, or lepidotrichia, which grow by successive addition of distal segments. The present study shows that evx1, a new zebrafish even-skipped related gene (Thaëron et al., 2000) displays during bony ray morphogenesis, a unique repetitive expression pattern along the proximodistal axis of the fin.

Expression of two even-skipped genes eve1 and evx2 during zebrafish fin morphogenesis and their regulation by retinoic acid.

Growth and patterning during fin regeneration depend, like for fin development, on the integrated expression of homeogenes. In the present work we have studied, by in situ hybridization, the expression and regulation of two vertebrate homologs eve1 and evx2 of the Drosophila pair-rule even-skipped gene family. Upon amputation of pectoral and caudal fins, both genes, expressed transiently in the mesenchyme during early stages of fin development of these fins, are turned on.

Involvement of the sonic hedgehog, patched 1 and bmp2 genes in patterning of the zebrafish dermal fin rays.

The signaling molecule encoded by Sonic hedgehog (shh) participates in the patterning of several embryonic structures including limbs. During early fin development in zebrafish, a subset of cells in the posterior margin of pectoral fin buds express shh. We have shown that regulation of shh in pectoral fin buds is consistent with a role in mediating the activity of a structure analogous to the zone of polarizing activity (ZPA) (Akimenko and Ekker (1995) Dev.

Gene expression during amphibian limb regeneration.

Limb regeneration in adult urodeles is an important phenomenon that poses fundamental questions both in biology and in medicine. In this review, we focus on recent advances in the characterization of the regeneration blastema at cellular and molecular levels and on the current understanding of the molecular basis of limb regeneration and its relationship to development. In particular, we discuss (i) the spatiotemporal distribution of genes and gene products in the mesenchyme and wound epidermis of the regenerating limb, (ii) how growth is controlled in the regeneration blastema, and (iii) molecules that are likely to be involved in patterning the regenerating limb such as homeobox genes and retinoids.

Correlation between RA-induced apoptosis and patterning defects in regenerating fins and limbs.

We have compared the ability of RA to induce apoptosis in regenerating fins and limbs in order to establish whether there may be a possible causal relationship between RA-induced cell death and the different patterning abnormalities observed in these two systems following RA treatment. In regenerating fins RA affects the anteroposterior axis and induces narrowing of the fin and fusion of rays. In regenerating limbs however, it mainly affects the proximodistal axis, where dose-dependent duplications of segments are observed.

Caudal fin regeneration in wild type and long-fin mutant zebrafish is affected by retinoic acid.

Zebrafish (Danio rerio) represents an ideal experimental model to tackle fundamental issues concerned with organogenesis during development and regeneration of complex body structures. We discuss here the development of the skeleton in zebrafish caudal fins, their regenerative ability in wild type and long-fin mutant adult fish, and how retinoic acid (RA), which induces duplications along the proximodistal axis in regenerating limbs, affects regeneration of the caudal fin. The dorsal and ventral lobes of zebrafish caudal fins are apparently symmetrical along the dorsoventral axis, but all of the skeletal elements and most of the soft tissues of both lobes originate from the ventral part of the embryo, as demonstrated by whole-mount staining of developing fish.

Retinoic acid-induced cell death in the wound epidermis of regenerating zebrafish fins.

Previous work has shown that treatment with retinoic acid (RA) can induce malformations in regenerating pectoral and caudal fins. RA-treated regenerates are narrower than unamputated and regenerated control fins because of a decrease in the distance between rays, and either partial or total fusion of some of them. In order to tackle the issue of how RA induces its teratogenic effects on regenerating fins, and which cell types may be specifically affected by RA, we have examined the cellular changes occurring in early regenerates following treatment with retinoids.

Formation of the peripheral nervous system during tail regeneration in urodele amphibians: ultrastructural and immunohistochemical studies of the origin of the cells.

In the regenerating newt tail, epimorphic regeneration--which recapitulates morphologically normal embryonic development--proceeds along a rostrocaudal differentiation gradient. Innervation of the new myomeres results from the spinal roots of segments rostral to the amputation plane and from ventral roots emerging from the lateroventral region of the regenerating spinal cord, in which motor neurons are differentiating. Electron microscopy and an indirect immunofluorescence study with anti-glial fibrillary acid protein (GFAP) confirm that the ventrolateral part of the regenerated ependymal tube gives rise to cells of the ventral root sheath and the spinal ganglia.

Nerve-dependent expression of c-myc protein during forelimb regeneration of Xenopus laevis froglets.

The consequences of denervation on the expression of c-myc protein have been analyzed on the regenerating forelimb of young froglets of Xenopus laevis. The level of c-myc expression, low in control limbs and enhanced in the regenerate, is transiently increased after a three-hour total denervation. For this protein, the level of expression is not a function of the quantity of nerve in the regenerate.

Nuclear distribution of PCNA during embryonic development in Xenopus laevis: a reinvestigation of early cell cycles.

The immunocytological distribution of the proliferating cell nuclear antigen (PCNA), a protein involved in DNA replication, has been examined during the early development of Xenopus laevis. The protein is uniformly detected in nuclei during early stages up to the neurula stage. PCNA is detected by its distinctive cyclical pattern during early development, remaining detectable only during the period of S phase of each cell cycle.

Organization and development of the mineral phase during early ontogenesis of the bony fin rays of the trout Oncorhynchus mykiss.

Characterization of mineral deposition has been studied by electron optical methods during early ontogenesis of lepidotrichia, the bony fin rays, of the trout Oncorhynchus mykiss (the former Salmo gairdneri). The fin rays consist of an extracellular granular ground substance containing in part a network of collagen fibrils within the basal lamella of the fin dermoepidermal interface. Growth of individual rays proceeds in a proximodistal direction.

Enhanced c-myc gene expression during forelimb regenerative outgrowth in the young Xenopus laevis.

Analysis of the expression of the c-myc protooncogene has been carried out in the forelimb regenerate of the Xenopus laevis froglet. Northern blot hybridization analysis revealed the presence of a 2.5-kilobase c-myc transcript in the regenerate forelimb at a level at least 7-fold more than the one found in nonregenerating forelimbs or stumps of regenerating forelimbs.

Genes and mechanisms involved in early embryonic development in Xenopus laevis.

Our laboratory is studying genes involved in the regulation of the balance between cell growth and differentiation during embryonic development in Xenopus. We have analyzed the developmental expression of the proto-oncogenes c-myc, and KiRas 2B, the proliferating cell nuclear antigen (PCNA), and the tumor suppressor gene p53. These genes, usually expressed during cell proliferation, are expressed in the oocyte in large quantities, but the majority of their maternal RNAs are degraded by the gastrula stage.

Early stages of spinal ganglion formation during tail regeneration in the newt, Notophthalmus viridescens.

Stages in the development of sensory ganglia in the regenerating newt tail after amputation are described by taking advantage of the rostrocaudal developmental gradient of the regenerating tail. A series of ganglia, beginning at the tip of the regenerate and progressing rostrally, were examined. Eight-week regenerates were used because they showed the most complete array of stages.

Fine structural peculiarities of the pectoral fin dermoskeleton of two brachiopterygii, Polypterus senegalus and Calamoichthys calabaricus (Pisces, Osteichthyes).

The features of the dermal skeleton of the pectoral fins of two Brachiopterygii, Polypterus senegalus and Calamoichthys calabaricus, have been studied by light and electron microscopy. The components studied are the ganoine-covered lepidotrichia segments and the distally located actinotrichia, the features of which are similar to those in teleosts. An irregular patch of ganoine susceptible to erosion by vascular canals lies on top of the cellular bone of the upper surface of the segment.

On the origin of ganoine: histological and ultrastructural data on the experimental regeneration of the scales of Calamoichthys calabaricus (Osteichthyes, Brachyopterygii, Polypteridae).

In order to understand the process of ganoine formation on the ganoid scales, scale regeneration has been studied to overcome the lack of a growth series of scale ontogeny. Seven stages of ganoid scale regeneration have been defined over a period of five months in the polypterid fish Calamoichthys calabaricus. The study has been carried out using transmission electron microscopic techniques.