Luis G. Morelli

Patterning embryos with oscillations: structure, function and dynamics of the vertebrate segmentation clock

The segmentation clock is an oscillating genetic network thought to govern the rhythmic and sequential subdivision of the elongating body axis of the vertebrate embryo into somites: the precursors of the segmented vertebral column. Understanding how the rhythmic signal arises, how it achieves precision and how it patterns the embryo remain challenging issues. Recent work has provided evidence of how the period of the segmentation clock is regulated and how this affects the anatomy of the embryo. The ongoing development of real-time clock reporters and mathematical models promise novel insight into the dynamic behavior of the clock.

Intercellular Coupling Regulates the Period of the Segmentation Clock

BACKGROUND Coupled biological oscillators can tick with the same period. How this collective period is established is a key question in understanding biological clocks. We explore this question in the segmentation clock, a population of coupled cellular oscillators in the vertebrate embryo that sets the rhythm of somitogenesis, the morphological segmentation of the body axis. The oscillating cells of the zebrafish segmentation clock are thought to possess noisy autonomous periods, which are synchronized by intercellular coupling through the Delta-Notch pathway. Here we ask whether Delta-Notch coupling additionally influences the collective period of the segmentation clock. RESULTS Using multiple-embryo time-lapse microscopy, we show that disruption of Delta-Notch intercellular coupling increases the period of zebrafish somitogenesis. Embryonic segment length and the spatial wavelength of oscillating gene expression also increase correspondingly, indicating an increase in the segmentation clocks period. Using a theory based on phase oscillators in which the collective period self-organizes because of time delays in coupling, we estimate the cell-autonomous period, the coupling strength, and the coupling delay from our data. Further supporting the role of coupling delays in the clock, we predict and experimentally confirm an instability resulting from decreased coupling delay time. CONCLUSIONS Synchronization of cells by Delta-Notch coupling regulates the collective period of the segmentation clock. Our identification of the first segmentation clock period mutants is a critical step toward a molecular understanding of temporal control in this system. We propose that collective control of period via delayed coupling may be a general feature of biological clocks.

Delayed coupling theory of vertebrate segmentation

Rhythmic and sequential subdivision of the elongating vertebrate embryonic body axis into morphological somites is controlled by an oscillating multicellular genetic network termed the segmentation clock. This clock operates in the presomitic mesoderm (PSM), generating dynamic stripe patterns of oscillatory gene‐expression across the field of PSM cells. How these spatial patterns, the clocks collective period, and the underlying cellular‐level interactions are related is not understood. A theory encompassing temporal and spatial domains of local and collective aspects of the system is essential to tackle these questions. Our delayed coupling theory achieves this by representing the PSM as an array of phase oscillators, combining four key elements: a frequency profile of oscillators slowing across the PSM; coupling between neighboring oscillators; delay in coupling; and a moving boundary describing embryonic axis elongation. This theory predicts that the segmentation clocks collective period depends on delayed coupling. We derive an expression for pattern wavelength across the PSM and show how this can be used to fit dynamic wildtype gene‐expression patterns, revealing the quantitative values of parameters controlling spatial and temporal organization of the oscillators in the system. Our theory can be used to analyze experimental perturbations, thereby identifying roles of genes involved in segmentation.

Topology and Dynamics of the Zebrafish Segmentation Clock Core Circuit

By combining biochemical, embryological, and mathematical approaches, this work uncovers an important role for protein-protein interactions in determining the dynamics of the somite-forming segmentation clock in vertebrates.

Self-propelled particles with fluctuating speed and direction of motion in two dimensions

We study general aspects of active motion with fluctuations in the speed and the direction of motion in two dimensions. We consider the case in which fluctuations in the speed are not correlated to fluctuations in the direction of motion, and assume that both processes can be described by independent characteristic time scales. We show the occurrence of a complex transient that can exhibit a series of alternating regimes of motion, for two different angular dynamics which correspond to persistent and directed random walks. We also show additive corrections to the diffusion coefficient. The characteristic time scales are also exposed in the velocity autocorrelation, which is a sum of exponential forms.

Computational Approaches to Developmental Patterning

Computational approaches are breaking new ground in understanding how embryos form. Here, we discuss recent studies that couple precise measurements in the embryo with appropriately matched modeling and computational methods to investigate classic embryonic patterning strategies. We include signaling gradients, activator-inhibitor systems, and coupled oscillators, as well as emerging paradigms such as tissue deformation. Parallel progress in theory and experiment will play an increasingly central role in deciphering developmental patterning.

A Doppler effect in embryonic pattern formation

Observing an embryonic Doppler effect The sound of an oncoming train changes as it passes you, a phenomenon termed the Doppler effect. Soroldoni et al. propose a similar event during the formation of vertebrate embryo body segments. It is generally thought that the internal timing of a genetic oscillator called the “segmentation clock” sets the rhythm of body segments called somites. However, time-lapse microscopy of the spatial waves of oscillations and the timing of body segment formation showed segments forming faster than spatial genetic oscillations. This “Doppler effect” occurs because the end of the oscillating tissue moves steadily into the oncoming waves. Thus, the rhythm of sequential body segmentation is a function of genetic oscillations, their changing wave pattern, and tissue shortening. Science, this issue p. 222 Genetic oscillations, their changing spatial pattern, and tissue shortening direct the rhythm of sequential body segmentation. During embryonic development, temporal and spatial cues are coordinated to generate a segmented body axis. In sequentially segmenting animals, the rhythm of segmentation is reported to be controlled by the time scale of genetic oscillations that periodically trigger new segment formation. However, we present real-time measurements of genetic oscillations in zebrafish embryos showing that their time scale is not sufficient to explain the temporal period of segmentation. A second time scale, the rate of tissue shortening, contributes to the period of segmentation through a Doppler effect. This contribution is modulated by a gradual change in the oscillation profile across the tissue. We conclude that the rhythm of segmentation is an emergent property controlled by the time scale of genetic oscillations, the change of oscillation profile, and tissue shortening.

Wnt-regulated dynamics of positional information in zebrafish somitogenesis

How signaling gradients supply positional information in a field of moving cells is an unsolved question in patterning and morphogenesis. Here, we ask how a Wnt signaling gradient regulates the dynamics of a wavefront of cellular change in a flow of cells during somitogenesis. Using time-controlled perturbations of Wnt signaling in the zebrafish embryo, we changed segment length without altering the rate of somite formation or embryonic elongation. This result implies specific Wnt regulation of the wavefront velocity. The observed Wnt signaling gradient dynamics and timing of downstream events support a model for wavefront regulation in which cell flow plays a dominant role in transporting positional information.

Persistence, period and precision of autonomous cellular oscillators from the zebrafish segmentation clock

In vertebrate development, the sequential and rhythmic segmentation of the body axis is regulated by a “segmentation clock”. This clock is comprised of a population of coordinated oscillating cells that together produce rhythmic gene expression patterns in the embryo. Whether individual cells autonomously maintain oscillations, or whether oscillations depend on signals from neighboring cells is unknown. Using a transgenic zebrafish reporter line for the cyclic transcription factor Her1, we recorded single tailbud cells in vitro. We demonstrate that individual cells can behave as autonomous cellular oscillators. We described the observed variability in cell behavior using a theory of generic oscillators with correlated noise. Single cells have longer periods and lower precision than the tissue, highlighting the role of collective processes in the segmentation clock. Our work reveals a population of cells from the zebrafish segmentation clock that behave as self-sustained, autonomous oscillators with distinctive noisy dynamics. DOI: http://dx.doi.org/10.7554/eLife.08438.001

Collective modes of coupled phase oscillators with delayed coupling.

We study the effects of delayed coupling on timing and pattern formation in spatially extended systems of dynamic oscillators. Starting from a discrete lattice of coupled oscillators, we derive a generic continuum theory for collective modes of long wavelengths. We use this approach to study spatial phase profiles of cellular oscillators in the segmentation clock, a dynamic patterning system of vertebrate embryos. Collective wave patterns result from the interplay of coupling delays and moving boundary conditions. We show that the phase profiles of collective modes depend on coupling delays.