In situ differentiation of iridophore crystallotypes underlies zebrafish stripe patterning.

Skin color patterns are ubiquitous in nature, impact social behavior, predator avoidance, and protection from ultraviolet irradiation. A leading model system for vertebrate skin patterning is the zebrafish; its alternating blue stripes and yellow interstripes depend on light-reflecting cells called iridophores. It was suggested that the zebrafish's color pattern arises from a single type of iridophore migrating differentially to stripes and interstripes. However, here we find that iridophores do not migrate between stripes and interstripes but instead differentiate and proliferate in-place, based on their micro-environment. RNA-sequencing analysis further reveals that stripe and interstripe iridophores have different transcriptomic states, while cryogenic-scanning-electron-microscopy and micro-X-ray diffraction identify different crystal-arrays architectures, indicating that stripe and interstripe iridophores are different cell types. Based on these results, we present an alternative model of skin patterning in zebrafish in which distinct iridophore crystallotypes containing specialized, physiologically responsive, organelles arise in stripe and interstripe by in-situ differentiation.

Gut-like ectodermal tissue in a sea anemone challenges germ layer homology.

Cnidarians (for example, sea anemones and jellyfish) develop from an outer ectodermal and inner endodermal germ layer, whereas bilaterians (for example, vertebrates and flies) additionally have a mesodermal layer as intermediate germ layer. Currently, cnidarian endoderm (that is, 'mesendoderm') is considered homologous to both bilaterian endoderm and mesoderm. Here we test this hypothesis by studying the fate of germ layers, the localization of gut cell types, and the expression of numerous 'endodermal' and 'mesodermal' transcription factor orthologues in the anthozoan sea anemone Nematostella vectensis. Surprisingly, we find that the developing pharyngeal ectoderm and its derivatives display a transcription-factor expression profile (foxA, hhex, islet, soxB1, hlxB9, tbx2/3, nkx6 and nkx2.2) and cell-type combination (exocrine and insulinergic) reminiscent of the developing bilaterian midgut, and, in particular, vertebrate pancreatic tissue. Endodermal derivatives, instead, display cell functions and transcription-factor profiles similar to bilaterian mesoderm derivatives (for example, somatic gonad and heart). Thus, our data supports an alternative model of germ layer homologies, where cnidarian pharyngeal ectoderm corresponds to bilaterian endoderm, and the cnidarian endoderm is homologous to bilaterian mesoderm.

Proliferation-independent regulation of organ size by Fgf/Notch signaling.

Organ morphogenesis depends on the precise orchestration of cell migration, cell shape changes and cell adhesion. We demonstrate that Notch signaling is an integral part of the Wnt and Fgf signaling feedback loop coordinating cell migration and the self-organization of rosette-shaped sensory organs in the zebrafish lateral line system. We show that Notch signaling acts downstream of Fgf signaling to not only inhibit hair cell differentiation but also to induce and maintain stable epithelial rosettes. Ectopic Notch expression causes a significant increase in organ size independently of proliferation and the Hippo pathway. Transplantation and RNASeq analyses revealed that Notch signaling induces apical junctional complex genes that regulate cell adhesion and apical constriction. Our analysis also demonstrates that in the absence of patterning cues normally provided by a Wnt/Fgf signaling system, rosettes still self-organize in the presence of Notch signaling.

Pre-bilaterian origin of the blastoporal axial organizer.

The startling capacity of the amphibian Spemann organizer to induce naïve cells to form a Siamese twin embryo with a second set of body axes is one of the hallmarks of developmental biology. However, the axis-inducing potential of the blastopore-associated tissue is commonly regarded as a chordate feature. Here we show that the blastopore lip of a non-bilaterian metazoan, the anthozoan cnidarian Nematostella vectensis, possesses the same capacity and uses the same molecular mechanism for inducing extra axes as chordates: Wnt/ß-catenin signaling. We also demonstrate that the establishment of the secondary, directive axis in Nematostella by BMP signaling is sensitive to an initial Wnt signal, but once established the directive axis becomes Wnt-independent. By combining molecular analysis with experimental embryology, we provide evidence that the emergence of the Wnt/ß-catenin driven blastopore-associated axial organizer predated the cnidarian-bilaterian split over 600 million years ago.

Wnt/ß-catenin dependent cell proliferation underlies segmented lateral line morphogenesis.

Morphogenesis is a fascinating but complex and incompletely understood developmental process. The sensory lateral line system consists of only a few hundred cells and is experimentally accessible making it an excellent model system to interrogate the cellular and molecular mechanisms underlying segmental morphogenesis. The posterior lateral line primordium periodically deposits prosensory organs as it migrates to the tail tip. We demonstrate that periodic proneuromast deposition is governed by a fundamentally different developmental mechanism than the classical models of developmental periodicity represented by vertebrate somitogenesis and early Drosophila development. Our analysis demonstrates that proneuromast deposition is driven by periodic lengthening of the primordium and a stable Wnt/ß-catenin activation domain in the leading region of the primordium. The periodic lengthening of the primordium is controlled by Wnt/ß-catenin/Fgf-dependent proliferation. Once proneuromasts are displaced into the trailing Wnt/ß-catenin-free zone they are deposited. We have previously shown that Wnt/ß-catenin signaling induces Fgf signaling and that interactions between these two pathways regulate primordium migration and prosensory organ formation. Therefore, by coordinating migration, prosensory organ formation and proliferation, localized activation of Wnt/ß-catenin signaling in the leading zone of the primordium plays a crucial role in orchestrating lateral line morphogenesis.

Cell-cell signaling interactions coordinate multiple cell behaviors that drive morphogenesis of the lateral line.

The zebrafish sensory lateral line system has emerged as a powerful model for the mechanistic study of collective cell migration and morphogenesis. Recent work has uncovered the details of a signaling network involving the Wnt/ß-catenin, Fgf and Delta-Notch pathways that patterns the migrating lateral line primordium into distinct regions. Cells within these regions exhibit different fundamental behaviors that together orchestrate normal lateral line morphogenesis. In this review, we summarize the signaling network that patterns the migrating lateral line primordium and describe how this patterning coordinates crucial morphogenic cell behaviors.

Cell migration during morphogenesis.

During development, functional structures must form with the correct three-dimensional geometry composed of the correct cell types. In many cases cell types are specified at locations distant to where they will ultimately reside for normal biological function. Although cell migration is crucial for normal development and morphogenesis of animal body plans and organ systems, abnormal cell migration during adult life underlies pathological states such as invasion and metastasis of cancer. In both contexts cells migrate either individually, as loosely associated sheets or as clusters of cells. In this review, we summarize, compare and integrate knowledge gained from several in vivo model systems that have yielded insights into the regulation of morphogenic cell migration, such as the zebrafish lateral line primordium and primordial germ cells, Drosophila border cell clusters, vertebrate neural crest migration and angiogenic sprouts in the post-natal mouse retina. Because of its broad multicontextual and multiphylletic distribution, understanding cell migration in its various manifestations in vivo is likely to provide new insights into both the function and malfunction of key embryonic and postembryonic events. In this review, we will provide a succinct phenotypic description of the many model systems utilized to study cell migration in vivo. More importantly, we will highlight, compare and integrate recent advances in our understanding of how cell migration is regulated in these varied model systems with special emphasis on individual and collective cell movements.

Multiple signaling interactions coordinate collective cell migration of the posterior lateral line primordium.

Collective migration of adherent cohorts of cells is a common and crucial phenomenon during embryonic development and adult tissue homeostasis. The zebrafish posterior lateral line primordium has emerged as a powerful in vivo model to study collective migration due to its relative simplicity and accessibility. While it has become clear that chemokine signaling is the primary guidance system responsible for directing the primordium along its migratory path it is not clear what mechanisms downstream of chemokine signaling coordinate migration of individual cells within the primordium. In this review, we summarize the cell signaling interactions that underlie collective migration of the primordium and discuss proposed mechanisms for the function of chemokine signaling in this tissue.

Wnt/beta-catenin and Fgf signaling control collective cell migration by restricting chemokine receptor expression.

Collective cell migration is a hallmark of embryonic morphogenesis and cancer metastases. However, the molecular mechanisms regulating coordinated cell migration remain poorly understood. A genetic dissection of this problem is afforded by the migrating lateral line primordium of the zebrafish. We report that interactions between Wnt/beta-catenin and Fgf signaling maintain primordium polarity by differential regulation of gene expression in the leading versus the trailing zone. Wnt/beta-catenin signaling in leader cells informs coordinated migration via differential regulation of the two chemokine receptors, cxcr4b and cxcr7b. These findings uncover a molecular mechanism whereby a migrating tissue maintains stable, polarized gene expression domains despite periodic loss of whole groups of cells. Our findings also bear significance for cancer biology. Although the Fgf, Wnt/beta-catenin, and chemokine signaling pathways are well known to be involved in cancer progression, these studies provide in vivo evidence that these pathways are functionally linked.