Morphogenetic Mechanisms Research Articles

D’Arcy Thompson and Synthetic Biology—Then and Now

AbstractThough often presented as a recent scientific endeavor, synthetic biology began in the 19th century and was a particularly active field in the years preceding the publication of D’Arcy Thompson’s On Growth and Form. Much synthetic biology of the era was devoted to the construction of nonliving chemical systems that would undergo morphogenesis or dynamic behaviors which had been observed in living organisms. The point was to show that “life-like” structure and behavior could be generated by physicochemical laws and required no vitalist element. D’Arcy Thompson’s careful analysis of physicochemical morphogenetic mechanisms as possible explanations of organic form links closely to this way of thinking. In the modern era, when we can genetically engineer cells to undergo specific behaviors, and program cells to undergo simple morphogenetic behaviors of the kind that Thompson and others felt might underly natural morphogenesis, it is possible to test whether they will in fact produce a predictable multicellular shape. This addresses essentially the same questions about the morphogenetic role of physicochemical forces, such as surface tension, but does so “the other way round”: physicochemical mechanisms are not being used as models for morphogenesis by natural cells but rather as a means to engineer cells to make designed forms.

The myoarchitecture of the vertebrate cardiac ventricles: evolution and classification.

The ventricle of the vertebrate heart is the main segment of the cardiac outflow region. Compared with other cardiac components, it shows remarkable histomorphological variation among different animal groups. This variation is especially apparent in the myocardium, which is generally classified into three main types: trabeculated, compact and mixed. The trabeculated or 'spongy' myocardium is characterized by the existence of trabeculae and deep recesses or intertrabecular spaces, lined by the endocardium. The compact type is composed of condensed myocardial fibers, with almost no trabeculated layer. The mixed type consists of an outer compact layer and an inner trabeculated layer. Among vertebrates, fishes show a great diversity of myocardial types. On this basis, the ventricular myoarchitecture has been categorized into four groups of varying complexity. This classification is made according to (i) the proportion of the two types of myocardium, trabeculated versus compact, and (ii) the vascularization of the heart wall. Here, we review the morphogenetic mechanisms that give rise to the different ventricular myoarchitecture in gnathostomes (i.e. jawed vertebrates) with special emphasis on the diversity of the ventricular myocardium throughout the phylogeny of ancient actinopterygians and teleosts. Finally, we propose that the classification of the ventricular myoarchitecture should be reconsidered, given that the degrees of myocardial compactness on which the current classification system is based do not constitute discrete states, but an anatomical continuum.

Analysis of the Structural Organization of the Human Finger Phalanges Using Anatomical Network Models

Despite the fact that the morphogenetic mechanisms of human finger transformation during ontogenesis are currently known, the issues of how they are organized into a complex integrated structure of the distal hand remains open. This question remains unanswered for several reasons, including the lack of consensus on conceptual definitions and approaches, as well as tools for assessing and comparing variations in several anatomical parts of the hand. The aim of the study was to investigate the structural organization of the human finger phalanges using anatomical network analysis (AnNA). Material and methods. In this study, the authors applied the IGRAPH package functions in the R data analysis programming environment for AnNA. Network modeling and layout were performed using the Fruchterman-Reingold algorithm. Analysis of the structure, as well as modularity and integration in the networks, was performed using the spin-glass algorithm. X-ray osteometric indices of the I–V fingers phalanx length were used to assess AnNA in 100 men and 100 women of the middle age without traumatic changes, deformations, and developmental deviations. Results. AnNA demonstrates a two-level organization of the distal hand in the form of a proximal module including the proximal phalanges and a distal module combining the middle and distal phalanges. When comparing the features of the network models of the distal hand, it was found that in women, the organization of the finger phalanges is characterised by higher morphological integration and modularity (modularity 0.43) than in men (modularity 0.38). orphological modularity and integration are organizing factors in the structure of the finger phalanges of the human distal hand. Conclusion. The study results demonstrate that the structural organization of the finger phalanges of the human hand is a system of individual anatomical modules of the phalanges.

Structure, function and formation of the amniote skin pattern

From feather and hair dotted arrays to pigmented stripes and spots, the spatial distribution of skin appendages and colouration often forms visible ornaments crucial for fitness in the coat of birds and mammals. These geometrical motifs are extremely diverse in nature. Yet, phenotypic surveys evidenced common themes in variation: the orientation, appendage-specificity or pigmentation of a given region may be conserved across groups or species. Here, we review naturalist observations of natural variation in the anatomy and ecological function of the skin pattern in amniotes. We then describe several decades of genetics, mathematical modelling and experimental embryology work aiming at understanding the molecular and morphogenetic mechanisms responsible for pattern formation. We discuss how these studies provided evidence that the morphological trends and differences representative of the phenotypic landscape of skin patterns in wild amniote species is rooted in the mechanisms controlling the production of distinct compartments in the embryonic skin.

An outer membrane porin-lipoprotein complex modulates elongasome movement to establish cell curvature in Rhodospirillum rubrum

Curved cell shapes are widespread among bacteria and important for cellular motility, virulence and fitness. However, the underlying morphogenetic mechanisms are still incompletely understood. Here, we identify an outer-membrane protein complex that promotes cell curvature in the photosynthetic species Rhodospirillum rubrum. We show that the R. rubrum porins Por39 and Por41 form a helical ribbon-like structure at the outer curve of the cell that recruits the peptidoglycan-binding lipoprotein PapS, with PapS inactivation, porin delocalization or disruption of the porin-PapS interface resulting in cell straightening. We further demonstrate that porin-PapS assemblies act as molecular cages that entrap the cell elongation machinery, thus biasing cell growth towards the outer curve. These findings reveal a mechanistically distinct morphogenetic module mediating bacterial cell shape. Moreover, they uncover an unprecedented role of outer-membrane protein patterning in the spatial control of intracellular processes, adding an important facet to the repertoire of regulatory mechanisms in bacterial cell biology.

The embryo-oil drop assembly: the timing and morphology of a critical event for fish early-life history survival

Egg specific gravity is of relevance for fish recruitment since the ability to float influences egg and larvae development, dispersal and connectivity between fishing grounds. Using zootechnics, histological approaches, optical and electronic transmission microscopy, this study describes the morphogenetic mechanism of adhesion of the oil-drop covering layer (OCL) to the oil droplet (OD) in embryos of Merluccius merluccius under physical conditions reflecting the marine environment. The herein described primordial (p)OCL is a substructure of the inner yolk syncytial layer which contains egg organella aimed to mobilize lipidic reserves from the oil drop (OD) towards the embryo blood. It is shown that the timely OD-OCL assembly is a critical morphogenetic process for embryo and larvae survival. Such assembly depends on egg buoyance because of its influence on the embryo capacity to rotate within the perivitelline space. Therefore, oil droplet adhesion (ODA) eggs are capable to complete their development while oil droplet non-adhesion eggs (ODNA) dye soon after hatching. We show that gravity-dependent egg buoyance categories exhibit different ODA/ODNA ratios (0–77%) and that relationship diminishes under incubation systems such as sprayers, that do not assure a dynamic seawater surface mixing to avoid egg desiccation. As an adaptive trait, egg gravity strongly depends on oceanic properties such as current dynamics, turbulence, oxygen, rainfall, and salinity, whose rapid changes would likely challenge the sustainability of fisheries recruitment.

Developmental process of the modern house shrew's molars: implications for the evolution of the tribosphenic molar in Mesozoic mammals.

Phylogenetically, the tribosphenic molars-prototypes of multi-cusped cheek teeth in marsupial and placental mammals-are derived from the single-cusped conical teeth of reptiles through the addition of cusps. Ontogenetically, mammalian molars are formed through the interface between the dental epithelium and mesenchyme (future enamel-dentin junction), becoming geometrically complex by adding epithelial signaling centers, called enamel knots, which determine future cusp positions. To reevaluate cusp homologies in Mesozoic mammals from an ontogenetic perspective, this study tracked molar development in a living placental mammal species, the house shrew (Suncus murinus), whose molars are morphologically the least derived from tribosphenic prototypes. The development of shrew molars proceeded as if it replayed the evolutionary process of tribosphenic molars. The first formed enamel knots gave rise to the evolutionarily oldest cusps-upper paracone and lower protoconid. The order of formation of other enamel knots and their location in development seemed to trace the order of cusp appearance in evolution. The parallel relationship between ontogeny and phylogeny of mammalian molars, if any, suggests that a change in the timing between developmental events rather than a change in the morphogenetic mechanism itself, should have been a major causal factor for the evolutionary transformation of tooth morphology.

Optical microscopic imaging, manipulation, and analysis methods for morphogenesis research.

Morphogenesis is a developmental process of organisms being shaped through complex and cooperative cellular movements. To understand the interplay between genetic programs and the resulting multicellular morphogenesis, it is essential to characterize the morphologies and dynamics at the single-cell level and to understand how physical forces serve as both signaling components and driving forces of tissue deformations. In recent years, advances in microscopy techniques have led to improvements in imaging speed, resolution and depth. Concurrently, the development of various software packages has supported large-scale, analyses of challenging images at the single-cell resolution. While these tools have enhanced our ability to examine dynamics of cells and mechanical processes during morphogenesis, their effective integration requires specialized expertise. With this background, this review provides a practical overview of those techniques. First, we introduce microscopic techniques for multicellular imaging and image analysis software tools with a focus on cell segmentation and tracking. Second, we provide an overview of cutting-edge techniques for mechanical manipulation of cells and tissues. Finally, we introduce recent findings on morphogenetic mechanisms and mechanosensations that have been achieved by effectively combining microscopy, image analysis tools and mechanical manipulation techniques.

Age-Related Changes in Sperm Morphology and Analysis of Multiple Sperm Defects.

Analysis of sperm morphology defects (amorphous heads, abnormal acrosome, etc.) is useful for estimating the efficiency of spermiogenesis and sperm maturation. An advanced paternal age (more than 40 years) is associated with decreasing sperm count and reduced motility; however, there is little information on the effect of aging relating to sperm morphological defects. Moreover, searching for stable combinations of certain morphological defects in the same sperm can be useful for better understanding spermiogenesis. The aim of the study was to investigate age-related changes in sperm morphology and the prevalence of certain combinations of sperm morphological defects in men from the general population. Sperm morphology was assessed in 1266 volunteers from the Russian urban general population in different age groups (18-19, 20-24, 25-29, 30-34, 35-40, and over 40 years old). Two hundred sperm were evaluated from each semen sample (about 250 thousand spermatozoa in total). Sperm defects were classified according to the WHO laboratory manual (WHO, 2010). The total percentage of each sperm defect and the frequency of different combinations of sperm morphological anomalies for each age group were counted. Additionally, a similar analysis was performed for the groups of normospermia and pathozoospermia. The frequency of coiled and short sperm tails increased in men over 40 years old compared to younger subjects; however, aging did not affect the percentage of morphologically normal sperm. It was shown that the combination of a misshaped head (amorphous, pyriform, and elongated) with a postacrosomal vacuole, acrosome defect, excess residual cytoplasm, or any anomaly of the midpiece or tail in the same spermatozoon were not random combinations of independent solitary defects. The increased frequency of combinations of coiled tails with amorphous, elongated, or vacuolated heads was observed in men older than 40 years. Sperm morphological defects, such as severely deformed heads (pyriform, elongated, and round) were more common in men with pathozoospermia compared to normospermic subjects. An age-related impairment in sperm morphology was found. Stable combinations of head defects with anomalies in the acrosome, midpiece or tail suggest that these defects may be the result of a general violation in the morphogenetic mechanism.

F-Actin Organization and Epidermal Cell Morphogenesis in the Brown Alga Sargassum vulgare.

The ordinary epidermal cells of various vascular plants are characterized by wavy anticlinal wall contours. This feature has not yet been reported in multicellular algal species. Here, we found that, in the leaf-like blades of the brown alga Sargassum vulgare, epidermal cells exhibit prominent waviness. Initially, the small meristodermal cells exhibit straight anticlinal contour, which during their growth becomes wavy, in a pattern highly reminiscent of that found in land plants. Waviness is restricted close to the external periclinal wall, while at inner levels the anticlinal walls become thick and even. The mechanism behind this shape relies on cortical F-actin organization. Bundles of actin filaments are organized, extending under the external periclinal wall and connecting its junctions with the anticlinal walls, constituting an interconnected network. These bundles define the sites of local thickening deposition at the anticlinal/periclinal wall junctions. These thickenings are interconnected by cellulose microfibril extensions under the external periclinal wall. Apart from the wavy anticlinal contour, outward protrusions also arise on the external periclinal wall, thus the whole epidermis exhibits a quilted appearance. Apart from highlighting a new role for F-actin in cell shaping, the comparison of this morphogenetic mechanism to that of vascular plants reveals a case of evolutionary convergence among photosynthetic organisms.

Mathematical and computational models in spheroid-based biofabrication.

Ubiquitous in embryonic development, tissue fusion is of interest to tissue engineers who use tissue spheroids or organoids as building blocks of three-dimensional (3D) multicellular constructs. This review presents mathematical models and computer simulations of the fusion of tissue spheroids. The motivation of this study stems from the need to predict the post-printing evolution of 3D bioprinted constructs. First, we provide a brief overview of differential adhesion, the main morphogenetic mechanism involved in post-printing structure formation. It will be shown that clusters of cohesive cells behave as an incompressible viscous fluid on the time scale of hours. The discussion turns then to mathematical models based on the continuum hydrodynamics of highly viscous liquids and on statistical mechanics. Next, we analyze the validity and practical use of computational models of multicellular self-assembly in live constructs created by tissue spheroid bioprinting. Finally, we discuss the perspectives of the field as machine learning starts to reshape experimental design, and modular robotic workstations tend to alleviate the burden of repetitive tasks in biofabrication. STATEMENT OF SIGNIFICANCE: Bioprinted constructs are living systems, which evolve via morphogenetic mechanisms known from developmental biology. This review presents mathematical and computational tools devised for modeling post-printing structure formation. They help achieving a desirable outcome without expensive optimization experiments. While previous reviews mainly focused on assumptions, technical details, strengths, and limitations of computational models of multicellular self-assembly, this article discusses their validity and practical use in biofabrication. It also presents an overview of mathematical models that proved to be useful in the evaluation of experimental data on tissue spheroid fusion, and in the calibration of computational models. Finally, the perspectives of the field are discussed in the advent of robotic biofabrication platforms and bioprinting process optimization by machine learning.

Immunohistochemical basing of the pathogenesis equality of varicose vein transformation in varicocele and varicose veins of the lower extremities

Background. Varicocele as well as varicose veins of the lower extremities are quite widespread. To date, the mechanisms of variceal transformation remain not completely clear.Aim. To study the immunohistochemical profile in order to substantiate the common pathogenesis of varicose veins transformation in varicocele and varicose veins of the lower limbs; to confirm the key role of undifferentiated connective tissue dysplasia syndrome in the development of these diseases.Materials and methods. Vein wall fragments from 24 male patients aged 6–35 years old divided into the corresponding groups: group 1–12 patients with varicocele, group 2–12 patients with varicose veins of the lower limbs. Control group – 5 practically healthy men (age 16–32 years). Immunohistochemical study was performed with monoclonal antibodies to collagen types III, IV, α-SMA, fibronectin, laminin, MMP-2 and MMP-9, TGF-β1.Results and discussion. Histological and immunohistochemical studies of vein wall biopsy specimens in both groups revealed similar morphological changes, different from the control group. When assessing the expression level of α-SMA in both study groups, there was marked expression in the bundles of hypertrophic smooth muscle cells (in the zone of wall thickening where fibrosis was minimal) and weak expression in the bundles of smooth muscle cells “enmeshed” in connective tissue sleeves. Collagen type III showed weak expression in the subintimal zone and between the bundles of superficially located smooth muscle cells in both study groups. Expression of type IV collagen was detected in the areas of wall thickening and hypertrophy of smooth muscle cells. In both groups there was a pronounced expression of fibronectin in the most altered sections of the vein walls, especially in the middle layer, around the bundles of smooth muscle cells. There was marked expression of MMP-2 in all layers of the wall, and overexpression of MMP-9 in the areas of wall thinning (in the subintimal zone and muscular layer). TGF-β1 overexpression was also detected in vein biopsy specimens from both groups, predominantly in areas of thinning and pronounced intermuscular fibrosis.Conclusion. Taking into account the similarity of immunohistochemical manifestations in the vein walls in varicocele and varicose veins of the lower limbs we can speak about the common morphogenetic mechanisms of their remodeling. Thus, we consider it possible to combine various forms of varicosity into a single disease – varicose vein disease.

Mutant IAA21 genes from Dendrocalamus sinicus Chia et J. L. Sun inhibit stem and root growth in transgenic tobacco by interacting with ARF5

Woody bamboos are important resource of industrial fibres. Auxin signaling plays a key role in multiple plant developmental processes, as yet the role of auxin/indole acetic acid (Aux/IAA) in culm development of woody bamboos has not been previously characterized. Dendrocalamus sinicus Chia et J. L. Sun is the largest woody bamboo documented in the world. Here, we identified two alleles of DsIAA21 gene (sIAA21 and bIAA21) from the straight- and bent-culm variants of D. sinicus, respectively, and studied how the domains I, i, and II of DsIAA21 affect the gene transcriptional repression. The results showed that bIAA21 expression was rapidly induced by exogenous auxin in D. sinicus. In transgenic tobacco, sIAA21 and bIAA21 mutated in domains i, and II significantly regulated plant architecture and root development. Stem cross sections revealed that parenchyma cells were smaller in transgenic plants than that in wild type plants. Domain i mutation changed the leucine and proline at position 45 to proline and leucine (siaa21L45P and biaa21P45L) strongly repressed cell expansion and root elongation by reducing the gravitropic response. Substitution of isoleucine with valine in domain II of the full length DsIAA21 resulted in dwarf stature in transgenic tobacco plants. Furthermore, the DsIAA21 interacted with auxin response factor 5 (ARF5) in transgenic tobacco plants, suggesting that DsIAA21 might inhibit stem and root elongation via interacting with ARF5. Taken together, our data indicated that DsIAA21 was a negative regulator of plant development and suggested that amino acid differences in domain i of sIAA21 versus bIAA21 affected their response to auxin, and might play a key role in the formation of the bent culm variant in D. sinicus. Our results not only shed a light on the morphogenetic mechanism in D. sinicus, but also provided new insights into versatile function of Aux/IAAs in plants.

Active cell divisions generate fourfold orientationally ordered phase in living tissue

Morphogenesis, the process through which genes generate form, establishes tissue-scale order as a template for constructing the complex shapes of the body plan. The extensive growth required to build these ordered substrates is fuelled by cell proliferation, which, naively, should destroy order. Understanding how active morphogenetic mechanisms couple cellular and mechanical processes to generate order—rather than annihilate it—remains an outstanding question in animal development. We show that cell divisions are the primary drivers of tissue flow, leading to a fourfold orientationally ordered phase. Waves of anisotropic cell proliferation propagate across the embryo with precise patterning. Defects introduced into the nascent lattice by cell divisions are moved out of the tissue bulk towards the boundary by subsequent divisions. Specific cell proliferation rates and orientations enable cell divisions to organize rather than fluidize the tissue. We observe this using live imaging and tissue cartography to analyse the dynamics of fourfold tissue ordering in the trunk segmental ectoderm of the crustacean Parhyale hawaiensis beginning 72 h after egg lay. The result is a robust, active mechanism for generating global orientational order in a non-equilibrium system that sets the stage for the subsequent development of shape and form.

Morphogenesis of the gastrovascular canal network in Aurelia jellyfish: Variability and possible mechanisms

Patterns in biology can be considered as predetermined or arising from a self-organizing instability. Variability in the pattern can, thus, be interpreted as a trace of instability, growing out from noise. Studying this variability can, thus, hint toward an underlying morphogenetic mechanism. Here, we present the variability of the gastrovascular system of the jellyfish Aurelia. In this variability emerges a typical biased reconnection between canals and time-correlated reconnections. Both phenomena can be interpreted as traces of mechanistic effects, the swimming contractions on the tissue surrounding the gastrovascular canals, and the mean fluid pressure inside them. This reveals the gastrovascular network as a model system to study the morphogenesis of circulation networks and the morphogenetic mechanisms at play.

Biocontrol of Meloidogyne spp. in Solanum lycopersicum using a dual combination of Bacillus strains.

Root-knot nematodes (RKNs, Meloidogyne spp.) are obligate plant parasites that constitute a significant pest for agriculture worldwide. They penetrate the plant roots, reducing the uptake of water and nutrients, causing a significant impact on crop yield. One alternative on focus now for nematode management is biological control. Rhizobacteria within the Bacillus genus show multiple modes of action against plant-parasitic nematodes (PPNs) that can act alone or in combination. In this context, we evaluated a dual-strain bacteria combination (B. paralicheniformi FMCH001 and B. subtilis FMCH002) to reduce nematode infection in tomato plants. We evaluated mortality of larvae from Meloidogyne javanica in vitro, as well as eggs hatching after the treatment. Atraction, penetration, establishment, and reproduction assays in vitro or in pots in tomato plants infected with M. javanica and treated/ untreated with the dual-strain bacteria combination were also performed. Additionally, morphometric parameters comparing giant cells size from galls of treated and untreated plants by using confocal microscopy were also measured. The results showed that this combination of strains has nematicidal properties in the pre-infection phase by decreasing the egg-hatching, juvenile survival, and attractiveness to the roots. Furthermore, nematode establishment, gall formation, and, remarkably, giant cell development was severely impaired after the bacterial treatment, suggesting interference with morphogenetic mechanisms induced by the nematode during GCs development within the plant. Nematode reproduction in tomato plants was reduced independently of the application mode in soil, before or after bacterial treatment. The dual-strain combination was also effective against other PPNs (i.e. Pratylenchus spp.) and in different crops (soybean). Therefore, combining B. paralicheniformis FMCH001 and B. subtilis FMCH002 is an efficient agent for the biological control of Meloidogyne spp. by interfering with different stages of the nematode cycle as a result of multiple modes of action.

Pyramidal neuron morphogenesis requires a septin network that stabilizes filopodia and suppresses lamellipodia during neurite initiation

Pyramidal neurons are a major cell type of the forebrain, consisting of a pyramidally shaped soma with axonal and apicobasal dendritic processes. It is poorly understood how the neuronal soma develops its pyramidal morphology, while generating neurites of the proper shape and orientation. Here, we discovered that the spherical somata of immature neurite-less neurons possess a circumferential wreath-like network of septin filaments, which promotes neuritogenesis by balancing the protrusive activity of lamellipodia and filopodia. In embryonic rat hippocampal and mouse cortical neurons, the septin wreath network consists of curvilinear filaments that contain septins 5, 7, and 11 (Sept5/7/11). The Sept5/7/11 wreath network demarcates a zone of myosin II enrichment and Arp2/3 diminution at the base of filopodial actin bundles. In Sept7-depleted neurons, cell bodies are enlarged with hyperextended lamellae and abnormally shaped neurites that originate from lamellipodia. This phenotype is accompanied by diminished myosin II and filopodia lifetimes and increased Arp2/3 and lamellipodial activity. Inhibition of Arp2/3 rescues soma and neurite phenotypes, indicating that the septin wreath network suppresses the extension of lamellipodia, facilitating the formation of neurites from the filopodia of a consolidated soma. We show that this septin function is critical for developing a pyramidally shaped soma with properly distributed and oriented dendrites in cultured rat hippocampal neurons and invivo in mouse perinatal cortical neurons. Therefore, the somatic septin cytoskeleton provides a key morphogenetic mechanism for neuritogenesis and the development of pyramidal neurons.

Double Puzzle: Morphogenesis of the Bi-Layered Leaf Adaxial Epidermis of Magnolia grandiflora.

Anticlinal ordinary epidermal cell wall waviness is a widespread feature found in the leaves of a variety of land plant species. However, it has not yet been encountered in leaves with multiple epidermides. Surprisingly, in Magnolia grandiflora leaves, ordinary epidermal cells in both layers of the bi-layered adaxial epidermis exhibit wavy anticlinal contour. During the development of the above cells, cortical microtubules are organized in anticlinally oriented bundles under the anticlinal walls, and radial arrays extending from the bundles at the edges of anticlinal and external periclinal walls, under the external periclinal walls. This microtubule pattern is followed by cell wall reinforcement with local thickenings, the cellulose microfibrils of which are parallel to the underlying microtubules. This specialized microtubule organization and concomitant cell wall reinforcement is initiated in the external epidermal layer, while hypodermis follows. The waviness pattern of each epidermal layer is unrelated to that of the other. The above findings are discussed in terms of morphogenetic mechanism induction and any implications in the functional significance of ordinary epidermal cell waviness.

Self-organizing morphogenetic mechanisms as processors of evolution.

Many organismic shapes and ornamentations are analogous to self-organizing patterns in the non-biological world. lt is suggested that responsible processes become first serendipitously "adopted" by developmental systems and then "lamed" by Darwinian selection as soon as a specific function has been acquired. Yet, taming can only reduce, bul not eliminare, the unpredictability of the dynamic systems at the local level.

A transient apical extracellular matrix relays cytoskeletal patterns to shape permanent acellular ridges on the surface of adult C. elegans.

Epithelial cells secrete apical extracellular matrices to form protruding structures such as denticles, ridges, scales, or teeth. The mechanisms that shape these structures remain poorly understood. Here, we show how the actin cytoskeleton and a provisional matrix work together to sculpt acellular longitudinal alae ridges in the cuticle of adult C. elegans. Transient assembly of longitudinal actomyosin filaments in the underlying lateral epidermis accompanies deposition of the provisional matrix at the earliest stages of alae formation. Actin is required to pattern the provisional matrix into longitudinal bands that are initially offset from the pattern of longitudinal actin filaments. These bands appear ultrastructurally as alternating regions of adhesion and separation within laminated provisional matrix layers. The provisional matrix is required to establish these demarcated zones of adhesion and separation, which ultimately give rise to alae ridges and their intervening valleys, respectively. Provisional matrix proteins shape the alae ridges and valleys but are not present within the final structure. We propose a morphogenetic mechanism wherein cortical actin patterns are relayed to the laminated provisional matrix to set up distinct zones of matrix layer separation and accretion that shape a permanent and acellular matrix structure.