The optic vesicle promotes cornea to lens transdifferentiation in larval Xenopus laevis

The outer cornea and pericorneal epidermis (lentogenic area) of larval Xenopus laevis are the only epidermal regions competent to regenerate a lens under the influence of the retinal inducer. However, the head epidermis of the lentogenic area can acquire the lens-regenerating competence following transplantation of an eye beneath it. In this paper we demonstrate that both the outer cornea and the head epidermis covering a transplanted eye are capable of responding not only to the retinal inducer of the larval eye but also to the inductive action of the embryonic optic vesicle by synthesizing crystallins. As the optic vesicle is a very weak lens inductor, which promotes crystallin synthesis only on the lens biased ectoderm of the embryo, these results indicate that the lens-forming competence in the outer cornea and epidermis of larval X. laevis corresponds to the persistence and acquisition of a condition similar to that of the embryonic biased ectoderm

Targeted expression of the dominant-negative FGFR4a in the eye using Xrx1A regulatory sequences interferes with normal retinal development

Molecular analysis of vertebrate eye development has been hampered by the availability of sequences that can selectively direct gene expression in the developing eye. We report the characterization of the regulatory sequences of the Xenopus laevis Rx1A gene that can direct gene expression in the retinal progenitor cells. We have used these sequences to investigate the role of Fibroblast Growth Factor (FGF) signaling in the development of retinal cell types. FGFs are signaling molecules that are crucial for correct patterning of the embryo and that play important roles in the development of several embryonic tissues. FGFs and their receptors are expressed in the developing retina, and FGF receptor-mediated signaling has been implicated to have a role in the specification and survival of retinal cell types. We investigated the role of FGF signaling mediated by FGF receptor 4a in the development of retinal cell types in Xenopus laevis. For this purpose, we have made transgenic Xenopus tadpoles in which the dominant-negative FGFR4a (ΔFGFR4a) coding region was linked to the newly characterized regulatory sequences of the Xrx1A gene. We found that the expression of ΔFGFR4a in retinal progenitor cells results in abnormal retinal development. The retinas of transgenic animals expressing ΔFGFR4a show disorganized cell layering and specifically lack photoreceptor cells. These experiments show that FGFR4a-mediated FGF signaling is necessary for the correct specification of retinal cell types. Furthermore, they demonstrate that constructs using Xrx1A regulatory sequences are excellent tools with which to study the developmental processes involved in retinal formation

The lens-regenerating competence in the outer cornea and epidermis of larval Xenopus laevis is related to pax6 expression.

After lentectomy, larval Xenopus laevis can regenerate a new lens by transdifferentiation of the outer cornea and pericorneal epidermis (lentogenic area). This process is promoted by retinal factor(s) accumulated into the vitreous chamber. To understand the molecular basis of the lens-regenerating competence (i.e. the capacity to respond to the retinal factor forming a new lens) in the outer cornea and epidermis, we analysed the expression of otx2, pax6, sox3, pitx3, prox1, betaB1-cry (genes all involved in lens development) by Real-time RT-PCR in the cornea and epidermis fragments dissected from donor larvae. The same fragments were also implanted into the vitreous chamber of host larvae to ascertain their lens-regenerating competence using specific anti-lens antibodies. The results demonstrate that there is a tight correlation between lens-regenerating competence and pax6 expression. In fact, (1) pax6 is the only one of the aforesaid genes to be expressed in the lentogenic area; (2) pax6 expression is absent in head epidermis outside the lentogenic area and in flank epidermis, both incapable of transdifferentiating into lens after implantation into the vitreous chamber; (3) in larvae that have undergone eye transplantation under the head or flank epidermis, pax6 re-expression was observed only in the head epidermis covering the transplanted eye. This is consistent with the fact that only the head epidermis reacquires the lens-regenerating competence after eye transplantation, forming a lens following implantation into the vitreous chamber; and (4) in larvae that have undergone removal of the eye, the epidermis covering the orbit maintained pax6 expression. This is consistent with the fact that after the eye enucleation the lentogenic area maintains the lens-regenerating competence, giving rise to a lens after implantation into the vitreous chamber. Moreover, we observed that misexpression of pax6 is sufficient to promote the acquisition of the lens-regenerating competence in flank epidermis. In fact, flank epidermis fragments dissected from pax6 RNA injected embryos could form lenses when implanted into the vitreous chamber. The data indicate for the first time that pax6 is a pivotal factor of lens-regenerating competence in the outer cornea and epidermis of larval X. laevis

Transcriptomic Analysis in the Sea Anemone Nematostella vectensis

International audienceThe sea anemone Nematostella vectensis is an emerging research model to study embryonic development and regeneration at the molecular and global transcriptomic level. Transcriptomics analysis is now routinely used to detect differential expression at the genome level. Here we present the latest procedures for isolating high-quality RNA required for next generation sequencing, as well as methods and resources for quantifying transcriptomic data