Vertebrate segmentation: from cyclic gene networks to scoliosis

One of the most striking features of the human vertebral column is its periodic organization along the anterior-posterior axis. This pattern is established when segments of vertebrates, called somites, bud off at a defined pace from the anterior tip of the embryo's presomitic mesoderm (PSM). To trigger this rhythmic production of somites, three major signaling pathways--Notch, Wnt/β-catenin, and fibroblast growth factor (FGF)--integrate into a molecular network that generates a traveling wave of gene expression along the embryonic axis, called the "segmentation clock." Recent systems approaches have begun identifying specific signaling circuits within the network that set the pace of the oscillations, synchronize gene expression cycles in neighboring cells, and contribute to the robustness and bilateral symmetry of somite formation. These findings establish a new model for vertebrate segmentation and provide a conceptual framework to explain human diseases of the spine, such as congenital scoliosis.     

Vertebrate neural stem cell segmentation, tracking and lineaging with validation and editing

This protocol and the accompanying software program called LEVER (lineage editing and validation) enable quantitative automated analysis of phase-contrast time-lapse images of cultured neural stem cells. Images are captured at 5-min intervals over a period of 5-15 d as the cells proliferate and differentiate. LEVER automatically segments, tracks and generates lineage trees of the stem cells from the image sequence. In addition to generating lineage trees capturing the population dynamics of clonal development, LEVER extracts quantitative phenotypic measurements of cell location, shape, movement and size. When available, the system can include biomolecular markers imaged using fluorescence. It then displays the results to the user for highly efficient inspection and editing to correct any errors in the segmentation, tracking or lineaging. To enable high-throughput inspection, LEVER incorporates features for rapid identification of errors and for learning from user-supplied corrections to automatically identify and correct related errors.     

Morph frequencies in the snail Cepaea nemoralis: changes with time and their interpretation

Colonies of the polymorphic snail Cepaea nemoralis on the sand dune at Point of Air, north Wales, first sampled in 1962 by A.J. Cain and P.M. Sheppard, have been re-sampled after 35 years. The frequency of the mid-banded morph has increased. Overall, yellow frequency has decreased at the expense of pink, but the change is in different directions in different parts of the dune. Yellow mid-banded and pink five-banded, phenotype combinations controlled by alleles at unlinked loci, are in excess within colonies, which suggests an effect of selection. It is argued that changes in morph frequency are due, at least in part, to movement between colonies. Cepaea populations are slow to respond to environmental changes and displacement of individuals may be greater than is often assumed. It is therefore difficult to interpret frequency changes over time. Correlated changes in groups of colonies may not always be interpretable as indicating selection.     

SOX3 activity during pharyngeal segmentation is required for craniofacial morphogenesis

Craniofacial development is a complex multi-step process leading to the morphogenesis of the face and sense organs, and to that of the neck, including the anteriormost part of the respiratory and digestive apparatus and associated endocrine glands. In vertebrates, the process is initiated by the formation of the pharyngeal arches from ectoderm, endoderm and mesoderm. These arches are then populated by neural crest cells, which originate from the central nervous system. We show here that, in mouse, there is a requirement for the HMG box factor SOX3 during the earliest stage of pharyngeal development: the formation of the pharyngeal pouches that segment the pharyngeal region by individualising each arch. In Sox3-null mutants, these pouches are expanded at the detriment of the second pharyngeal arch. As a consequence, neural crest cell migration and ectoderm-derived epibranchial placode development are affected, leading to craniofacial defects. We also show that Sox3 genetically interacts both with FgfR1 and with Sox2, another member of the Soxb1 family, to fulfil its function in the pharyngeal region. Although the importance of the neural crest has long been recognised, our studies highlight the equally crucial role of the pharyngeal region in craniofacial morphogenesis. They also give insight into the formation of pharyngeal pouches, of which little is known in vertebrates. Finally, this work introduces two new players in craniofacial development - SOX3 and SOX2.     

Oscillations of the snail genes in the presomitic mesoderm coordinate segmental patterning and morphogenesis in vertebrate somitogenesis

The segmented body plan of vertebrate embryos arises through segmentation of the paraxial mesoderm to form somites. The tight temporal and spatial control underlying this process of somitogenesis is regulated by the segmentation clock and the FGF signaling wavefront. Here, we report the cyclic mRNA expression of Snail 1 and Snail 2 in the mouse and chick presomitic mesoderm (PSM), respectively. Whereas Snail genes' oscillations are independent of NOTCH signaling, we show that they require WNT and FGF signaling. Overexpressing Snail 2 in the chick embryo prevents cyclic Lfng and Meso 1 expression in the PSM and disrupts somite formation. Moreover, cells mis-expressing Snail 2 fail to express Paraxis, remain mesenchymal, and are thereby inhibited from undergoing the epithelialization event that culminates in the formation of the epithelial somite. Thus, Snail genes define a class of cyclic genes that coordinate segmentation and PSM morphogenesis.     

Developmental modularity and phenotypic novelty within a biphasic life cycle: morphogenesis of a cone snail venom gland

The venom gland of predatory cone snails (Conus spp.), which secretes neurotoxic peptides that rapidly immobilize prey, is a proposed key innovation for facilitating the extraordinary feeding behaviour of these gastropod molluscs. Nevertheless, the unusual morphology of this gland has generated controversy about its evolutionary origin and possible homologues in other gastropods. I cultured feeding larvae of Conus lividus and cut serial histological sections through the developing foregut during larval and metamorphic stages to examine the development of the venom gland. Results support the hypothesis of homology between the venom gland and the mid-oesophageal gland of other gastropods. They also suggest that the mid-region of the gastropod foregut, like the anterior region, is divisible into dorsal and ventral developmental modules that have different morphological, functional and ontogenetic fates. In larvae of C. lividus, the ventral module of the middle foregut transformed into the anatomically novel venom gland of the post-metamorphic stage by rapidly pinching-off from the main dorsal channel of the mid-oesophagus, an epithelial remodelling process that may be similar to other cases where epithelial tubes and vesicles arise from a pre-existing epithelial sheet. The developmental remodelling mechanism could have facilitated an abrupt evolutionary transition to the derived morphology of this important gastropod feeding innovation.     

Vertebrate gastrulation: the BMP sticker shock

BMPs are essential regulators of cell fate during early embryonic development. Molecular genetics and in vivo imaging of cell behaviors in zebrafish now demonstrate a role for BMPs in the control of cell adhesion. The work reveals an important new mechanism governing cell movements during gastrulation.     

Vertebrate development: taming the nodal waves

Recent results have provided evidence that regulatory loops operating in vertebrate embryos between the developmental signalling factors Nodal and Lefty may provide a real example of the kind of reaction-diffusion process long predicted to be a mechanism of pattern formation.     

Vertebrate evolution: turning heads

The skeleton of the neck and shoulders has undergone alterations during evolution, but muscle connectivity has not. A recent study suggests this is a result of neural crest cells defining attachment points and thus muscle connectivity.     

Vertical movement and time allocation of a freshwater pulmonate snail

The paper presents an analysis of locomotion and location in a vertical water column of 10Biomphalaria glabrata, under constant conditions of light, temperature, and food availability. Individual snails varied in distance traveled, and in the percentage of time spent in different areas of the water column. Distance covered ranged from 53 cm to 100 cm h^–1 . Approximately 21% of locomotion was observed to be passive (floating up or sinking down). Snails spent approximately 58% of the time in the bottom section, 35% in the top section and only 7% in the middle region. The study provides baseline data that can be used to compare further studies ofBiomphalaria glabrata as various conditions are altered.