The physical basis of mollusk shell chiral coiling.

Snails are model organisms for studying the genetic, molecular, and developmental bases of left-right asymmetry in Bilateria. However, the development of their typical helicospiral shell, present for the last 540 million years in environments as different as the abyss or our gardens, remains poorly understood. Conversely, ammonites typically have a bilaterally symmetric, planispiraly coiled shell, with only 1% of 3,000 genera displaying either a helicospiral or a meandering asymmetric shell. A comparative analysis suggests that the development of chiral shells in these mollusks is different and that, unlike snails, ammonites with asymmetric shells probably had a bilaterally symmetric body diagnostic of cephalopods. We propose a mathematical model for the growth of shells, taking into account the physical interaction during development between the soft mollusk body and its hard shell. Our model shows that a growth mismatch between the secreted shell tube and a bilaterally symmetric body in ammonites can generate mechanical forces that are balanced by a twist of the body, breaking shell symmetry. In gastropods, where a twist is intrinsic to the body, the same model predicts that helicospiral shells are the most likely shell forms. Our model explains a large diversity of forms and shows that, although molluscan shells are incrementally secreted at their opening, the path followed by the shell edge and the resulting form are partly governed by the mechanics of the body inside the shell, a perspective that explains many aspects of their development and evolution.

Morphomechanics and Developmental Constraints in the Evolution of Ammonites Shell Form.

The idea that physical processes involved in biological development underlie morphogenetic rules and channel morphological evolution has been central to the rise of evolutionary developmental biology. Here, we explore this idea in the context of seashell morphogenesis. We show that a morphomechanical model predicts the effects of variations in shell shape on the ornamental pattern in ammonites, a now extinct group of cephalopods with external chambered shell. Our model shows that several seemingly unrelated characteristics of synchronous, ontogenetic, intraspecific, and evolutionary variations in ornamental patterns among various ammonite species may all be understood from the fact that the mechanical forces underlying the oscillatory behavior of the shell secreting system scale with the cross-sectional curvature of the shell aperture. This simple morphogenetic rule, emerging from biophysical interactions during shell formation, introduced a non-random component in the production of phenotypic variation and channeled the morphological evolution of ammonites over millions of years. As such, it provides a paradigm for the concept of "developmental constraints."

Cope's Rule and the Universal Scaling Law of Ornament Complexity.

Luxuriant, bushy antlers, bizarre crests, and huge, twisting horns and tusks are conventionally understood as products of sexual selection. This view stems from both direct observation and from the empirical finding that the size of these structures grows faster than body size (i.e., ornament size shows positive allometry). We contend that the familiar evolutionary increase in the complexity of ornaments over time in many animal clades is decoupled from ornament size evolution. Increased body size comes with extended growth. Since growth scales to the quarter power of body size, we predicted that ornament complexity should scale according to the quarter power law as well, irrespective of the role of sexual selection in the evolution and function of the ornament. To test this hypothesis, we selected three clades (ammonites, deer, and ceratopsian dinosaurs) whose species bore ornaments that differ in terms of the importance of sexual selection to their evolution. We found that the exponent of the regression of ornament complexity to body size is the same for the three groups and is statistically indistinguishable from 0.25. We suggest that the evolution of ornament complexity is a by-product of Cope's rule. We argue that although sexual selection may control size in most ornaments, it does not influence their shape.

Transitions during cephalopod life history: the role of habitat, environment, functional morphology and behaviour.

Cephalopod life cycles generally share a set of stages that take place in different habitats and are adapted to specific, though variable, environmental conditions. Throughout the lifespan, individuals undertake a series of brief transitions from one stage to the next. Four transitions were identified: fertilisation of eggs to their release from the female (1), from eggs to paralarvae (2), from paralarvae to subadults (3) and from subadults to adults (4). An analysis of each transition identified that the changes can be radical (i.e. involving a range of morphological, physiological and behavioural phenomena and shifts in habitats) and critical (i.e. depending on environmental conditions essential for cohort survival). This analysis underlines that transitions from eggs to paralarvae (2) and from paralarvae to subadults (3) present major risk of mortality, while changes in the other transitions can have evolutionary significance. This synthesis suggests that more accurate evaluation of the sensitivity of cephalopod populations to environmental variation could be achieved by taking into account the ontogeny of the organisms. The comparison of most described species advocates for studies linking development and ecology in this particular group.

A Na+ channel receptor of FMRFamide in the cephalopod Sepiella japonica: Identification, characterisation, and expression profiling during different stages of gonadal development.

FMRFamide, a member of the neuropeptide family, is involved in numerous physiological processes. FMRFamide-activated sodium channels (FaNaCs) are a family of non-voltage-gated, amiloride-sensitive, Na+-selective channels triggered by the neuropeptide FMRFamide. In the present study, the full-length cDNA of the FaNaC receptor of Sepiella japonica (SjFaNaC) was cloned. The cDNA of SjFaNaC was 3004 bp long with an open reading frame (ORF) of 1812 bp, encoding 603 amino acid residues with no signal peptide at the N-terminus. Sequence analysis indicated that SjFaNaC shared a high identity with other cephalopods FaNaCs and formed a sister clade with bivalves. The protein structure was predicted using SWISS-MODEL with AcFaNaC as the template. Quantitative real-time PCR (qRT-PCR) revealed that SjFaNaC transcripts were highly expressed in both female and male reproductive organs, as well as in the optic lobe and brain of the central nervous system (CNS). Results of in situ hybridisation (ISH) showed that SjFaNaC mRNA was mainly distributed in the medulla and deep retina of the optic lobe and in both the supraesophageal and subesophageal masses of the brain. Subcellular localisation indicated that the SjFaNaC protein was localised intracellularly and on the cell surface of HEK293T cells. In summary, these findings may lay the foundation for future exploration of the functions of SjFaNaC in cephalopods.

Quantification of ontogenetic allometry in ammonoids.

Ammonoids are well-known objects used for studies on ontogeny and phylogeny, but a quantification of ontogenetic change has not yet been carried out. Their planispirally coiled conchs allow for a study of "longitudinal" ontogenetic data, that is data of ontogenetic trajectories that can be obtained from a single specimen. Therefore, they provide a good model for ontogenetic studies of geometry in other shelled organisms. Using modifications of three cardinal conch dimensions, computer simulations can model artificial conchs. The trajectories of ontogenetic allometry of these simulations can be analyzed in great detail in a theoretical morphospace. A method for the classification of conch ontogeny and quantification of the degree of allometry is proposed. Using high-precision cross-sections, the allometric conch growth of real ammonoids can be documented and compared. The members of the Ammonoidea show a wide variety of allometric growth, ranging from near isometry to monophasic, biphasic, or polyphasic allometry. Selected examples of Palaeozoic and Mesozoic ammonoids are shown with respect to their degree of change during ontogeny of the conch.

[Ontophylogenetic studies of Paleozoic ammonoides].

In this work the main directions of study of ontogeny and phylogeny of Paleozoic ammonoids are discussed, and the results of studies of the Permian families of this subclass are presented. It is shown that the morphogenetic evolution of taxa of different rank is caused by manifestation of major phylogenetic moduses and their various combinations. Thus, development can proceed in the direction of morphological complexity and in the direction of simplification. The conclusion about the role of family in the evolutionary history of ammonoids, which depends on the complexity of its structure, was made: the more complex it is, the greater the perspective taxon can be for formation of new groups of the supraspecific rank.

Ontogeny, evolution and palaeogeographic distribution of the world's largest ammonite Parapuzosia (P.) seppenradensis (Landois, 1895).

The world's largest ammonite, Parapuzosia (P.) seppenradensis (Landois, 1895), fascinated the world ever since the discovery, in 1895, of a specimen of 1.74 metres (m) diameter near Seppenrade in Westfalia, Germany, but subsequent findings of the taxon are exceedingly rare and its systematic position remains enigmatic. Here we revise the historical specimens and document abundant new material from England and Mexico. Our study comprises 154 specimens of large (< 1 m diameter) to giant (> 1m diameter) Parapuzosia from the Santonian and lower Campanian, mostly with stratigraphic information. High-resolution integrated stratigraphy allows for precise cross-Atlantic correlation of the occurrences. Our specimens were analysed regarding morphometry, growth stages and stratigraphic occurrence wherever possible. Our analysis provides insight into the ontogeny of Parapuzosia (P.) seppenradensis and into the evolution of this species from its potential ancestor P. (P.) leptophylla Sharpe, 1857. The latter grew to shell diameters of about 1 m and was restricted to Europe in the early Santonian, but it reached the Gulf of Mexico during the late Santonian. P. (P.) seppenradensis first appears in the uppermost Santonian- earliest Campanian on both sides of the Atlantic. Initially, it also reached diameters of about 1 m, but gradual evolutionary increase in size is seen in the middle early Campanian to diameters of 1.5 to 1.8 m. P. (P.) seppenradensis is characterized by five ontogenetic growth stages and by size dimorphism. We therefore here include the many historic species names used in the past to describe the morphological and size variability of the taxon. The concentration of adult shells in small geographic areas and scarcity of Parapuzosia in nearby coeval outcrop regions may point to a monocyclic, possibly even semelparous reproduction strategy in this giant cephalopod. Its gigantism exceeds a general trend of size increase in late Cretaceous cephalopods. Whether the coeval increase in size of mosasaurs, the top predators in Cretaceous seas, caused ecological pressure on Parapuzosia towards larger diameters remains unclear.

Spatial distribution of oncocerid cephalopods on a Middle Devonian bedding plane suggests semelparous life cycle.

Reproductive strategies of extinct organisms can only be recognised indirectly and hence, they are exceedingly rarely reported and tend to be speculative. Here, we present a mass-occurrence with common preservation of pairs of late Givetian (Middle Devonian) oncocerid cephalopods from Hamar Laghdad in the Tafilalt (eastern Anti-Atlas, Morocco). We analysed their spatial occurrences with spatial point pattern analysis techniques and Monte Carlo simulations; our results shows that the pairwise clustering is significant, while ammonoids on the same bedding plane reveal a more random distribution. It is possible that processes such as catastrophic mass mortality or post-mortem transport could have produced the pattern. However, we suggest that it is more likely that the oncocerids were semelparous and died shortly after mating. These findings shed new light on the variation and evolution of reproductive strategies in fossil cephalopods and emphasise that they cannot be based on comparisons with extant taxa without question.

Cephalopod Biology: At the Intersection Between Genomic and Organismal Novelties.

Cephalopods are resourceful marine predators that have fascinated generations of researchers as well as the public owing to their advanced behavior, complex nervous system, and significance in evolutionary studies. Recent advances in genomics have accelerated the pace of cephalopod research. Many traditional areas focusing on evolution, development, behavior, and neurobiology, primarily on the morphological level, are now transitioning to molecular approaches. This review addresses the recent progress and impact of genomic and other molecular resources on research in cephalopods. We outline several key directions in which significant progress in cephalopod research is expected and discuss its impact on our understanding of the genetic background behind cephalopod biology and beyond.

Chamber volume development, metabolic rates, and selective extinction in cephalopods.

Reconstructing the physiology of extinct organisms is key to understanding mechanisms of selective extinction during biotic crises. Soft tissues of extinct organisms are rarely preserved and, therefore, a proxy for physiological aspects is needed. Here, we examine whether cephalopod conchs yield information about their physiology by assessing how the formation of chambers respond to external stimuli such as environmental changes. We measured chamber volume through ontogeny to detect differences in the pattern of chamber volume development in nautilids, coleoids, and ammonoids. Results reveal that the differences between ontogenetic trajectories of these cephalopods involve the presence or absence of abrupt decreases of chamber volume. Accepting the link between metabolic rate and growth, we assume that this difference is rooted in metabolic rates that differ between cephalopod clades. High metabolic rates combined with small hatching size in ammonoids as opposed to lower metabolic rates and much larger hatchlings in most nautilids may explain the selective extinction of ammonoids as a consequence of low food availability at the end of the Cretaceous.

Ecological pressures and the contrasting scaling of metabolism and body shape in coexisting taxa: cephalopods versus teleost fish.

Metabolic rates are fundamental to many biological processes, and commonly scale with body size with an exponent ( bR) between 2/3 and 1 for reasons still debated. According to the 'metabolic-level boundaries hypothesis', bR depends on the metabolic level ( LR). We test this prediction and show that across cephalopod species intraspecific bR correlates positively with not only LR but also the scaling of body surface area with body mass. Cephalopod species with high LR maintain near constant mass-specific metabolic rates, growth and probably inner-mantle surface area for exchange of respiratory gases or wastes throughout their lives. By contrast, teleost fish show a negative correlation between bR and LR. We hypothesize that this striking taxonomic difference arises because both resource supply and demand scale differently in fish and cephalopods, as a result of contrasting mortality and energetic pressures, likely related to different locomotion costs and predation pressure. Cephalopods with high LR exhibit relatively steep scaling of growth, locomotion, and resource-exchange surface area, made possible by body-shape shifting. We suggest that differences in lifestyle, growth and body shape with changing water depth may be useful for predicting contrasting metabolic scaling for coexisting animals of similar sizes. This article is part of the theme issue 'Physiological diversity, biodiversity patterns and global climate change: testing key hypotheses involving temperature and oxygen'.

Evolution of limb development in cephalopod mollusks.

Cephalopod mollusks evolved numerous anatomical novelties, including arms and tentacles, but little is known about the developmental mechanisms underlying cephalopod limb evolution. Here we show that all three axes of cuttlefish limbs are patterned by the same signaling networks that act in vertebrates and arthropods, although they evolved limbs independently. In cuttlefish limb buds, Hedgehog is expressed anteriorly. Posterior transplantation of Hedgehog-expressing cells induced mirror-image limb duplications. Bmp and Wnt signals, which establish dorsoventral polarity in vertebrate and arthropod limbs, are similarly polarized in cuttlefish. Inhibition of Bmp2/4 dorsally caused ectopic expression of Notum, which marks the ventral sucker field, and ectopic sucker development. Cuttlefish also show proximodistal regionalization of Hth, Exd, Dll, Dac, Sp8/9, and Wnt expression, which delineates arm and tentacle sucker fields. These results suggest that cephalopod limbs evolved by parallel activation of a genetic program for appendage development that was present in the bilaterian common ancestor.

Thorson's rule, life-history evolution, and diversification of benthic octopuses (Cephalopoda: Octopodoidea).

Here, we evaluate the so-called Thorson's rule, which posits that direct-development and larger eggs are favored toward the poles in marine organisms and whose validity been the subject of considerable debate in the literature, combining an expanded phenotypic dataset encompassing 60 species of benthic octopuses with a new molecular phylogeny. Phylogenetic reconstruction shows two clades: clade 1 including species of the families Eledonidae, Megaleledonidae, Bathypolypodidae, and Enteroctopodidae, and clade 2 including species of Octopodidae. Egg size, development mode, and all environmental variables exhibited phylogenetic signal, partly due to differences between the two clades: whereas most species in clade 1 inhabit cold and deep waters, exhibit large eggs and hatchling with holobenthic development, species from clade 2 inhabit tropical-temperate and shallow waters, evolved small eggs, and generally exhibit merobenthic development. Phylogenetic regressions show that egg size exhibits a conspicuous latitudinal cline, and that both egg size and development mode vary with water temperature. Additionally, analyses suggest that egg size is constrained by body size in lineages with holobenthic development. Taken together, results suggest that the variation in egg size and development mode across benthic octopuses is adaptive and associated with water temperature, supporting Thorson's rule in these organisms.

Southern Ocean cephalopods.

The Southern Ocean cephalopod fauna is distinctive, with high levels of endemism in the squid and particularly in the octopodids. Loliginid squid, sepiids and sepiolids are absent from the Southern Ocean, and all the squid are oceanic pelagic species. The octopodids dominate the neritic cephalopod fauna, with high levels of diversity, probably associated with niche separation. In common with temperate cephalopods, Southern Ocean species appear to be semelparous, but growth rates are probably lower and longevity greater than temperate counterparts. Compared with equivalent temperate species, eggs are generally large and fecundity low, with putative long development times. Reproduction may be seasonal in the squid but is extended in the octopodids. Cephalopods play an important role in the ecology of the Southern Ocean, linking the abundant mesopelagic fish and crustaceans with higher predators such as albatross, seals and whales. To date Southern Ocean cephalopods have not been commercially exploited, but there is potential for exploitation of muscular species of the Family Ommastrephidae.

Early Mode of Life and Hatchling Size in Cephalopod Molluscs: Influence on the Species Distributional Ranges.

Cephalopods (nautiluses, cuttlefishes, squids and octopuses) exhibit direct development and display two major developmental modes: planktonic and benthic. Planktonic hatchlings are small and go through some degree of morphological changes during the planktonic phase, which can last from days to months, with ocean currents enhancing their dispersal capacity. Benthic hatchlings are usually large, miniature-like adults and have comparatively reduced dispersal potential. We examined the relationship between early developmental mode, hatchling size and species latitudinal distribution range of 110 species hatched in the laboratory, which represent 13% of the total number of live cephalopod species described to date. Results showed that species with planktonic hatchlings reach broader distributional ranges in comparison with species with benthic hatchlings. In addition, squids and octopods follow an inverse relationship between hatchling size and species latitudinal distribution. In both groups, species with smaller hatchlings have broader latitudinal distribution ranges. Thus, squid and octopod species with larger hatchlings have latitudinal distributions of comparatively minor extension. This pattern also emerges when all species are grouped by genus (n = 41), but was not detected for cuttlefishes, a group composed mainly of species with large and benthic hatchlings. However, when hatchling size was compared to adult size, it was observed that the smaller the hatchlings, the broader the latitudinal distributional range of the species for cuttlefishes, squids and octopuses. This was also valid for all cephalopod species with benthic hatchlings pooled together. Hatchling size and associated developmental mode and dispersal potential seem to be main influential factors in determining the distributional range of cephalopods.

Paleobiology of the Latest Tithonian (Late Jurassic) Ammonite Salinites grossicostatum Inferred from Internal and External Shell Parameters.

Based on material from the uppermost Tithonian La Caja Formation at Puerto Piñones, northeastern Mexico, the complete ontogenetic development (protoconch to adult) of the ammonite Salinites grossicostatum is outlined by a detailed morphometrical shell analysis. The embryonic stage, consisting of a small ellipsoid protoconch and ammonitella, ends at about 0.6 mm. Four major morphological changes are differentiated throughout ontogeny based on internal features such as reduced septal spacing and siphuncle position. Sexual dimorphism is reflected by shell size, siphuncular diameter, differences in the morphology of the apophysis, and by two distinct general trends in septal spacing. In addition, macroconchs are characterized by septal crowding at different stages, followed by the return to normal septum distances. Our analysis indicates a change in the mode of life after the neanic stage. A change in habitat preference is inferred for adult individuals. While microconchs persisted at Puerto Piñones, large mature macroconchs temporarily migrated to other areas, possibly for egg deposition. Salinites grossicostatum is endemic to the ancient Gulf of Mexico and is there restricted to outer continental shelf environments.

Contrasting Responses to Harvesting and Environmental Drivers of Fast and Slow Life History Species.

According to their main life history traits, organisms can be arranged in a continuum from fast (species with small body size, short lifespan and high fecundity) to slow (species with opposite characteristics). Life history determines the responses of organisms to natural and anthropogenic factors, as slow species are expected to be more sensitive than fast species to perturbations. Owing to their contrasting traits, cephalopods and elasmobranchs are typical examples of fast and slow strategies, respectively. We investigated the responses of these two contrasting strategies to fishing exploitation and environmental conditions (temperature, productivity and depth) using generalized additive models. Our results confirmed the foreseen contrasting responses of cephalopods and elasmobranchs to natural (environment) and anthropogenic (harvesting) influences. Even though a priori foreseen, we did expect neither the clear-cut differential responses between groups nor the homogeneous sensitivity to the same factors within the two taxonomic groups. Apart from depth, which affected both groups equally, cephalopods and elasmobranchs were exclusively affected by environmental conditions and fishing exploitation, respectively. Owing to its short, annual cycle, cephalopods do not have overlapping generations and consequently lack the buffering effects conferred by different age classes observed in multi-aged species such as elasmobranchs. We suggest that cephalopods are sensitive to short-term perturbations, such as seasonal environmental changes, because they lack this buffering effect but they are in turn not influenced by continuous, long-term moderate disturbances such as fishing because of its high population growth and turnover. The contrary would apply to elasmobranchs, whose multi-aged population structure would buffer the seasonal environmental effects, but they would display strong responses to uninterrupted harvesting due to its low population resilience. Besides providing empirical evidence to the theoretically predicted contrasting responses of cephalopods and elasmobranchs to disturbances, our results are useful for the sustainable exploitation of these resources.

The Evolution and Development of Cephalopod Chambers and Their Shape.

The Ammonoidea is a group of extinct cephalopods ideal to study evolution through deep time. The evolution of the planispiral shell and complexly folded septa in ammonoids has been thought to have increased the functional surface area of the chambers permitting enhanced metabolic functions such as: chamber emptying, rate of mineralization and increased growth rates throughout ontogeny. Using nano-computed tomography and synchrotron radiation based micro-computed tomography, we present the first study of ontogenetic changes in surface area to volume ratios in the phragmocone chambers of several phylogenetically distant ammonoids and extant cephalopods. Contrary to the initial hypothesis, ammonoids do not possess a persistently high relative chamber surface area. Instead, the functional surface area of the chambers is higher in earliest ontogeny when compared to Spirula spirula. The higher the functional surface area the quicker the potential emptying rate of the chamber; quicker chamber emptying rates would theoretically permit faster growth. This is supported by the persistently higher siphuncular surface area to chamber volume ratio we collected for the ammonite Amauroceras sp. compared to either S. spirula or nautilids. We demonstrate that the curvature of the surface of the chamber increases with greater septal complexity increasing the potential refilling rates. We further show a unique relationship between ammonoid chamber shape and size that does not exist in S. spirula or nautilids. This view of chamber function also has implications for the evolution of the internal shell of coleoids, relating this event to the decoupling of soft-body growth and shell growth.

Adaptation of the Mitochondrial Genome in Cephalopods: Enhancing Proton Translocation Channels and the Subunit Interactions.

Mitochondrial protein-coding genes (mt genes) encode subunits forming complexes of crucial cellular pathways, including those involved in the vital process of oxidative phosphorylation (OXPHOS). Despite the vital role of the mitochondrial genome (mt genome) in the survival of organisms, little is known with respect to its adaptive implications within marine invertebrates. The molluscan Class Cephalopoda is represented by a marine group of species known to occupy contrasting environments ranging from the intertidal to the deep sea, having distinct metabolic requirements, varied body shapes and highly advanced visual and nervous systems that make them highly competitive and successful worldwide predators. Thus, cephalopods are valuable models for testing natural selection acting on their mitochondrial subunits (mt subunits). Here, we used concatenated mt genes from 17 fully sequenced mt genomes of diverse cephalopod species to generate a robust mitochondrial phylogeny for the Class Cephalopoda. We followed an integrative approach considering several branches of interest-covering cephalopods with distinct morphologies, metabolic rates and habitats-to identify sites under positive selection and localize them in the respective protein alignment and/or tridimensional structure of the mt subunits. Our results revealed significant adaptive variation in several mt subunits involved in the energy production pathway of cephalopods: ND5 and ND6 from Complex I, CYTB from Complex III, COX2 and COX3 from Complex IV, and in ATP8 from Complex V. Furthermore, we identified relevant sites involved in protein-interactions, lining proton translocation channels, as well as disease/deficiencies related sites in the aforementioned complexes. A particular case, revealed by this study, is the involvement of some positively selected sites, found in Octopoda lineage in lining proton translocation channels (site 74 from ND5) and in interactions between subunits (site 507 from ND5) of Complex I.