Genomics reveal ancient forms of stanniocalcin in amphioxus and tunicate

Stanniocalcin (STC) is present throughout vertebrates, including humans, but a structure for STC has not been identified in animals that evolved before bony fish. The origin of this pleiotropic hormone known to regulate calcium is not clear. In the present study, we have cloned three stanniocalcins from two invertebrates, the tunicate Ciona intestinalis and the amphioxus Branchiostoma floridae. Both species are protochordates with the tunicates as the closest living relatives to vertebrates. Amphioxus are basal to both tunicates and vertebrates. The genes and predicted proteins of tunicate and amphioxus share several key structural features found in all previously described homologs. Both the invertebrate and vertebrate genes have four conserved exons. The predicted length of the single pro-STC in Ciona is 237 amino acids and the two pro-hormones in amphioxus are 207 and 210 residues, which is shorter than human pro-STCs at 247 and 302 residues due to expansion of the C-terminal region in vertebrate forms. The conserved pattern of 10 cysteines in all chordate STCs is crucial for identification as amphioxus and tunicate amino acids are only 14-23% identical with human STC1 and STC2. The 11th cysteine, which is the cysteine shown to form a homodimer in vertebrates, is present only in amphioxus STCa, but not in amphioxus STCb or tunicate STC, suggesting the latter two are monomers. The expression of stanniocalcin in Ciona is widespread as shown by RT-PCR and by quantitative PCR. The latter method shows that the highest amount of STC mRNA is in the heart with lower amounts in the neural complex, branchial basket, and endostyle. A widespread distribution is present also in mammals and fish for both STC1 and STC2. Stanniocalcin is a presumptive regulator of calcium in both Ciona and amphioxus, although the structure of a STC receptor remains to be identified in any organism. Our data suggest that amphioxus STCa is most similar to the common ancestor of vertebrate STCs because it has an 11th cysteine necessary for dimerization, an N-glycosylation motif, although not the canonical one in vertebrate STCs, and similar gene organization. Tunicate and amphioxus STCs are more similar in structure to vertebrate STC1 than to vertebrate STC2. The unique features of STC2, including 14 instead of 11 cysteines and a cluster of histidines in the C-terminal region, appear to be found exclusively in vertebrates.     

Segmentation of the Vertebrate Head

Historical views of head segmentation are reviewed. The concensusis that the head is segmented essentially in terms of myomeres,and that other organs have responded in varying degrees to this.From the various lines of reasoning a model of the primitivevertebrate is generated. This model denies the tunicate originof the vertebrates—rather it identifies amphioxus as mostlike the ancestral vertebrate. The vertebrate head is made upof a preoral segment plus four other segments. Because of sclerotomites,the head extends through five and a half segments. The nasalorgans and eyes are preoral structures while the ear is locatedbetween segments three and four. The occipital portion of thehead skeleton is formed from the posterior half of the fifthsegment and the anterior half of the sixth; it is vertebra-likein structure. This "segment" is much altered as a result ofthe multiplication of the visceral pouches and is often viewedas the fusion product of several segments. Thus the idea ofcorrespondence between somite and visceral segments posteriorto the second branchial arch is rejected. In some fishes, additionalvertebrae are added to the posterior part of the cranium andthis can be observed in development. The bony cranium of thevertebrate appears to partially reflect segmentation; its componentssuggest a vertebra-like developmental influence in operation.Study of the shark head has contributed much to our knowledgeof this area.     

Exploring developmental, functional, and evolutionary aspects of amphioxus sensory cells

Amphioxus has neither elaborated brains nor definitive sensory organs, so that the two may have evolved in a mutually affecting manner and given rise to the forms seen in extant vertebrates. Clarifying the developmental and functional aspects of the amphioxus sensory system is thus pivotal for inferring the early evolution of vertebrates. Morphological studies have identified and classified amphioxus sensory cells; however, it is completely unknown whether the morphological classification makes sense in functional and evolutionary terms. Molecular markers, such as gene expression, are therefore indispensable for investigating the developmental and functional aspects of amphioxus sensory cells. This article reviews recent molecular studies on amphioxus sensory cells. Increasing evidence shows that the non-neural ectoderm of amphioxus can be subdivided into molecularly distinct subdomains by the combinatorial code of developmental cues involving the RA-dependent Hox code, suggesting that amphioxus epithelial sensory cells developed along positional information. This study focuses particularly on research involving the molecular phylogeny and expression of the seven-transmembrane, G protein-coupled receptor (GPCR) genes and discusses the usefulness of this information for characterizing the sensory cells of amphioxus.     

Evolutionary Patterns of Axial Muscle Systems in Some Invertebrates and Fish

SYNOPSIS. Axial muscles used for oscillatory swimming are foundnot only in fish and other vertebrates but also in some protochordatesand invertebrates. Chaetognaths have unsegmented locomotor musculaturewith some unusual features, but larvacean tunicates and thetadpole larvae of ascidians show the simplest variant of thechordate segmented axial muscle arrangement for flexing a notochordalcolumn, where all muscle cells along one side are electricallycoupled. With amphioxus, the basic fish myotomal layout is established,with two main fibre types probably used for different patternsof swimming (as in fish). There are, however, several uniquefeatures, including the flattened fibre shape and the paramyosinsystem of the notochord. Agnatha have two fibre types in themyotomes, a third type perhaps being a developmental stage inthe ontogeny of fast fibres. In lampreys, the central fibresof the characteristic fibre sandwiches in the myotomes are flattened(though less so than in amphioxus); they have a dual innervationof unknown function seen also in the fast fibre system of manyGnathostome fish groups. Hagfish fast fibres are not flattenednor do they have a dual innervation. Gnathostome fish axialmuscles are strikingly uniform in design with two possible exceptions:(1) higher teleost fast fibres which, unlike those of othergroups, are multiply-innervated and (2) tonic fibres in a fewfish, which seem not to be involved in locomotion.     

Risk‐taking and the evolution of mechanisms for rapid escape from predators

Flight initiation distance (FID) is the distance at which an individual animal takes flight when approached by a human. This behavioural measure of risk‐taking reflects the risk of being captured by real predators, and it correlates with a range of life history traits, as expected if flight distance optimizes risk of predation. Given that FID provides information on risk of predation, we should expect that physiological and morphological mechanisms that facilitate flight and escape predict interspecific variation in flight distance. Haematocrit is a measure of packed red blood cell volume and as such indicates the oxygen transport ability and hence the flight muscle contracting reaction of an individual. Therefore, we predicted that species with short flight distances, that allow close proximity between a potential prey individual and a predator, would have high haematocrit. Furthermore, we predicted that species with large wing areas and hence relatively low costs of flight and species with large aspect ratios and hence high manoeuvrability would have evolved long flight speed. Consistent with these predictions, we found in a sample of 63 species of birds that species with long flight distances for their body size had low levels of haematocrit and large wing areas and aspect ratios. These findings provide evidence consistent with the evolution of risk‐taking behaviour being underpinned by physiological and morphological mechanisms that facilitate escape from predators and add to our understanding of predator–prey coevolution.     

The amphioxus SoxB family: implications for the evolution of vertebrate placodes

Cranial placodes are regions of thickened ectoderm that give rise to sense organs and ganglia in the vertebrate head. Homologous structures are proposed to exist in urochordates, but have not been found in cephalochordates, suggesting the first chordates lacked placodes. SoxB genes are expressed in discrete subsets of vertebrate placodes. To investigate how placodes arose and diversified in the vertebrate lineage we isolated the complete set of SoxB genes from amphioxus and analyzed their expression in embryos and larvae. We find that while amphioxus possesses a single SoxB2 gene, it has three SoxB1 paralogs. Like vertebrate SoxB1 genes, one of these paralogs is expressed in non-neural ectoderm destined to give rise to sensory cells. When considered in the context of other amphioxus placode marker orthologs, amphioxus SoxB1 expression suggests a diversity of sensory cell types utilizing distinct placode-type gene programs was present in the first chordates. Our data supports a model for placode evolution and diversification whereby the full complement of vertebrate placodes evolved by serial recruitment of distinct sensory cell specification programs to anterior pre-placodal ectoderm.     

Predator–prey interactions,flight initiation distance and brain size

Prey avoid being eaten by assessing the risk posed by approaching predators and responding accordingly. Such an assessment may result in prey–predator communication and signalling, which entail further monitoring of the predator by prey. An early antipredator response may provide potential prey with a selective advantage, although this benefit comes at the cost of disturbance in terms of lost foraging opportunities and increased energy expenditure. Therefore, it may pay prey to assess approaching predators and determine the likelihood of attack before fleeing. Given that many approaching potential predators are detected visually, we hypothesized that species with relatively large eyes would be able to detect an approaching predator from afar. Furthermore, we hypothesized that monitoring of predators by potential prey relies on evaluation through information processing by the brain. Therefore, species with relatively larger brains for their body size should be better able to monitor the intentions of a predator, delay flight for longer and hence have shorter flight initiation distances than species with smaller brains. Indeed, flight initiation distances increased with relative eye size and decreased with relative brain size in a comparative study of 107 species of birds. In addition, flight initiation distance increased independently with size of the cerebellum, which plays a key role in motor control. These results are consistent with cognitive monitoring as an antipredator behaviour that does not result in the fastest possible, but rather the least expensive escape flights. Therefore, antipredator behaviour may have coevolved with the size of sense organs, brains and compartments of the brain involved in responses to risk of predation.     

The role of the notochard in forward and reverse swimming and burrowing in the amphioxus Branchiostoma lanceolatum

Analyses of high speed cinefilm have shown that amphioxus swims either forward or backward with undulatory movement generated at the leading end, the wave of displacement passing along the body with increasing amplitude. The leading end, whether this is "head" or tail, is evidently more rigid than the trailing end, flexibility at each end changing with reversal in direction of swimming. It is suggested that control of the amplitude of the waves of displacement in different regions of the body in swimming is a function of the notochord, contraction of the muscular notochordal plates increasing its stiffness. Connections between the central nervous system and the notochordal plates via the notochordal pits are already known to exist.
As exposure to light invariably induces swimming in dark–adapted animals, it seems probable that the eyes function in initiating movement. The rate of increase in number and size of the eye cups during larval and adult growth and their pattern of distribution in the nerve cord are given. In the adult the eye cups occur predominantly in the anterior and posterior regions of the body. This may be of significance in providing the stimulus for changes in flexibility of these regions in swimming.
High speed cinefilm has also shown that amphioxus can burrow "head" or tail-first and move through sand in a forward or a reverse direction. It is suggested that rapid reversal of direction is of greater importance in movement through sand than in swimming.     

Diving in head first: trade‐offs between phenotypic traits and sand‐diving predator escape strategy in Meroles desert lizards

Survival, in part, depends on an individual's ability to evade predators. In desert regions some lizard species have evolved head‐first sand‐diving strategies to escape predators. To facilitate this behaviour, a distinctive head morphology that facilitates sand‐diving has evolved. This specialised head morphology may, however, come at a cost to other ecologically relevant functions, particularly bite force. Here, we investigated the relationship between morphology and function in a southern African lacertid lizard genus, Meroles, which consists of eight species that utilise different escape strategies, including sand‐diving and running for cover. It was hypothesized that the specialised head morphology of diving species would negatively affect bite force capacity. We found that species from each escape strategy category differed significantly in head shape, but not bite force performance. A phylogenetic tree of the genus was constructed using two mitochondrial and two nuclear genes, and we conducted phylogenetic comparative analyses. One aspect of the head shape differed between the escape strategies once phylogeny was taken into account. We found that bite force may have co‐evolved with head morphology, but that there was no trade‐off between biting capacity and escape strategy in Meroles.     

Evolution of prey behavior in response to changes in predation regime: damselflies in fish and dragonfly lakes

In a large behavioral experiment we reconstructed the evolution of behavioral responses to predators to explore how interactions with predators have shaped the evolution of their prey's behavior. All Enallagma damselfly species reduced both movement and feeding in the presence of coexisting predators. Some Enallagma species inhabit water bodies with both fish and dragonflies, and these species responded to the presence of both predators, whereas other Enallagma species inhabit water bodies that have only large dragonflies as predators, and these species only responded to the presence of dragonflies. Lineages that shifted to live with large dragonflies showed no evolution in behaviors expressed in the presence of dragonflies, but they evolved greater movement in the absence of predators and greater movement and feeding in the presence of fish. These results suggest that Enallagma species have evolutionarily lost the ability to recognize fish as a predator. Because species coexisting with only dragonfly predators have also evolved the ability to escape attacking dragonfly predators by swimming, the decreased predation risk associated with foraging appears to have shifted the balance of the foraging/predation risk trade-off to allow increased activity in the absence of mortality threats to evolve in these lineages. Our results suggest that evolution in response to changes in predation regime may have greater consequences for characters expressed in the absence of mortality threats because of how the balance between the conflicting demands of growth and predation risk are altered.     

Functional mechanisms of an inducible defence in tadpoles: morphology and behaviour influence mortality risk from predation

In many amphibian larvae a suite of morphological and behavioural characters varies together in an induced defence against predators, but it remains unclear which features are functionally related to defence. We independently manipulated behaviour and morphology in tadpoles of Hyla versicolor and assessed their consequences for swimming performance and predator escape. Data on burst swimming showed that tadpoles which accelerated rapidly were elongate, with shallow bodies and tails. Predator escape was measured by exposing tadpoles to predators (larval Anax dragonflies or larval Ambystoma salamanders) and recording time until death. Tadpoles were first reared for 30 days in ponds containing either caged Anax or no predators; individuals responded to predators by developing large brightly coloured tails and short bodies. We placed tadpoles of both morphological phenotypes into plastic tubs, and manipulated their behaviour using food and chemical cues from predators. Mortality risk experienced by the predator‐induced phenotype was about half that of the no‐predator phenotype, and risk increased with time spent swimming. An interaction between morphology and behaviour arose because increasing activity caused higher risk for tadpoles with deep tail fins but not shallow tail fins.     

A new calcichordate from the Ordovician of Bohemia and its anatomy, adaptations and relationships

The paper describes a new member of a group of Lower Palaeozoic marine fossils which partly bridge the gap between echinoderms and chordates. Evidence suggests that this group included the ancestors of the vertebrates. Its members are traditionally regarded as primitive echinoderms, but are better seen as primitive chordates with echinoderm affinities. They form a basal subphylum of chordates-the Calcichordata Jefferies 1967. The Calcichordata, in accordance with an early suggestion by Gislén, are probably ancestral to all living chordates. The new calcichordate is named Reticulocarpos hanusi gen et sp. nov. It comes from the Lower Ordovician ?árka Formation (Llanvirn) of ?árka near Prague, Czechoslovakia and is placed in the family Amygdalothecidae Ubaghs 1970. It is important because of its position in the Calcichordata. This group is divided into two very different orders–the Cornuta and the Mitrata. The Cornuta are the more primitive order and gave rise to the Mitrata, which had the structure of giant, calcite-plated tunicate tadpoles. Many features show that the new species is a very advanced cornute, closely related to the stock that gave rise to the mitrates. For this reason it is important in the general history of the chordates, since some primitive mitrate was probably the latest common ancestor of the living chordate subphyla i.e. of tunicates, of amphioxus and its allies and of the vertebrates. Being a mitrate-like cornute, the new species allows the cornutes and mitrates to be compared more confidently than before. Four results are especially important. Firstly it is likely that the stem (=tail) of mitrates is equivalent only to the anterior part of the stem of cornutes. This is significant, because traditional views as to which was the upper surface in mitrates have been based on stem homologies now seen as false. Secondly Reticulocarpos hanusi is adapted to stay up on very soft mud, using only the strength of the mud for support. The mitrates, on the other hand, supported themselves on soft mud by a much more reliable method resembling buoyancy. Thirdly, the new form had paired transpharyngeal eyes which are otherwise known only in mitrates, and which are the earliest type of paired eyes in chordates. Fourthly, it becomes possible to homologize the thecal plates of cornutes with those of mitrates. Reticulocarpos hanusi represents an important phase in chordate evolution dominated by the necessity of staying up on mud by a very precarious method. During this phase many pre-adaptations for swimming were acquired. Primitive mitrates, descended from a very similar form, were probably the first chordates that could swim.     

Costs and Benefits of Opisthobranch Swimming and Neurobehavioral Mechanisms

After opisthobranch molluscs dislodge from the substrate duringonset of swimming, the ensuing flexion or undulatory motionsare usually not well oriented with respect to predators, preyor suitable substrate. Swimming motions are effective in launchinganimals off the substrate and elevating them into the watercolumn where they are primarily transported passively by ambientwaves and tidal currents. Both active swimming and passive transporton ambient currents may provide escape from predators, searchfor food and mates, and dispersal to new and potentially adaptivelocations. However, loss of contact with the substrate and launchinginto the water column may also bring a high cost in terms ofexposure to diverse risks. I illustrate several forms of opisthobranchswimming and describe their mechanisms and roles. In addition,adaptations of some opisthobranchs to reduce the risks of exposureto predators during swimming are suggested. These adaptationsinclude small size, transparency or inconspicuous color to reducepredation while swimming, and neurobehavioral mechanisms ofrheotaxis and geomagnetic sensitivity.     

Opposite patterns of diurnal activity in the box jellyfish Tripedalia cystophora and Copula sivickisi

Cubozoan medusae have a stereotypic set of 24 eyes, some of which are structurally similar to vertebrate and cephalopod eyes. Across the approximately 25 described species, this set of eyes varies surprisingly little, suggesting that they are involved in an equally stereotypic set of visual tasks. During the day Tripedalia cystophora is found at the edge of mangrove lagoons where it accumulates close to the surface in sun-lit patches between the prop roots. Copula sivickisi (formerly named Carybdea sivickisi) is associated with coral reefs and has been observed to be active at night. At least superficially, the eyes of the two species are close to identical. We studied the diurnal activity pattern of these two species both in the wild and under controlled conditions in laboratory experiments. Despite the very similar visual systems, we found that they display opposite patterns of diurnal activity. T. cystophora is active exclusively during the day, whereas C. sivickisi is actively swimming at night, when it forages and mates. At night T. cystophora is found on the muddy bottom of the mangrove lagoon. C. sivickisi spends the day attached to structures such as the underside of stones and coral skeletons. This species difference seems to have evolved to optimize foraging, since the patterns of activity follow those of the available prey items in their respective habitats.     

Central neural circuitry in the jellyfish Aglantha: a model 'simple nervous system'

Like other hydrozoan medusae, Aglantha lacks a brain, but the two marginal nerve rings function together as a central nervous system. Twelve neuronal and two excitable epithelial conduction systems are described and their interactions summarized. Aglantha differs from most medusae in having giant axons. It can swim and contract its tentacles in two distinct ways (escape and slow). Escape responses are mediated primarily by giant axons but conventional interneurons are also involved in transmission of information within the nerve rings during one form of escape behavior. Surprisingly, giant axons provide the motor pathway to the swim muscles in both escape and slow swimming. This is possible because these axons can conduct calcium spikes as well as sodium spikes and do so on an either/or basis without overlap. The synaptic and ionic bases for these responses are reviewed. During feeding, the manubrium performs highly accurate flexions to points at the margin. At the same time, the oral lips flare open. The directional flexions are conducted by FMRFamide immunoreactive nerves, the lip flaring by an excitable epithelium lining the radial canals. Inhibition of swimming during feeding is due to impulses propagated centrifugally in the same epithelium. Aglantha probably evolved from an ancestor possessing a relatively simple wiring plan, as seen in other hydromedusae. Acquisition of giant axons resulted in considerable modification of this basic plan, and required novel solutions to the problems of integrating escape with non-escape circuitry.     

Interacting effects of predation risk and resource level on escape speed of amphibian larvae along a latitudinal gradient

Fast‐growing genotypes living in time‐constrained environments are often more prone to predation, suggesting that growth‐predation risk trade‐offs are important factors maintaining variation in growth along climatic gradients. However, the mechanisms underlying how fast growth increases predation‐mediated mortality are not well understood. Here, we investigated if slow‐growing, low‐latitude individuals have faster escape swimming speed than fast‐growing high‐latitude individuals using common frog (Rana temporaria) tadpoles from eight populations collected along a 1500 km latitudinal gradient. We measured escape speed in terms of burst and endurance speeds in tadpoles raised in the laboratory at two food levels and in the presence and absence of a predator (Aeshna dragonfly larvae). We did not find any latitudinal trend in escape speed performance. In low food treatments, burst speed was higher in tadpoles reared with predators but did not differ between high‐food treatments. Endurance speed, on the contrary, was lower in high‐food tadpoles reared with predators and did not differ between treatments at low food levels. Tadpoles reared with predators showed inducible morphology (increased relative body size and tail depth), which had positive effects on speed endurance at low but not at high food levels. Burst speed was positively affected by tail length and tail muscle size in the absence of predators. Our results suggest that escape speed does not trade‐off with fast growth along the latitudinal gradient in R. temporaria tadpoles. Instead, escape speed is a plastic trait and strongly influenced by the interaction between resource level and predation risk.     

Predation,cover, and convergent evolution in epipelagic oceans

The factor of most importance to the structure of epipelagic oceanic communities is the absence of cover and the inability to hide from predators in surface waters during the day (Elton, 1939). Visual predation in an environment devoid of cover has resulted in convergent evolution into only six modal adaptive patterns. Large, fast, visual predators roam the water, 1) alone or in 2) schools, and they eat anything of appropriate size that they see. Prey escape only by dint of 3) very small size, 4) invisibility due to tissue transparency, 5) diurnal vertical migration, or by 6) exploitation of the sea surface. The sensory ecology and physiology of zooplankton are different from that of all other animal categories in all other habitats. Epipelagic zooplankton are either extremely small animals, with small and structurally simple sense organs, or they are large, with gelatinous, transparent bodies which often lack sense organs.     

A Gbx homeobox gene in amphioxus: insights into ancestry of the ANTP class and evolution of the midbrain/hindbrain boundary

In the vertebrate central nervous system (CNS), mutual antagonism between posteriorly expressed Gbx2 and anteriorly expressed Otx2 positions the midbrain/hindbrain boundary (MHB), but does not induce MHB organizer genes such as En, Pax2/5/8 and Wnt1. In the CNS of the cephalochordate amphioxus, Otx is also expressed anteriorly, but En, Pax2/5/8 and Wnt1 are not expressed near the caudal limit of Otx, raising questions about the existence of an MHB organizer in amphioxus. To investigate the evolutionary origins of the MHB, we cloned the single amphioxus Gbx gene. Fluorescence in situ hybridization showed that, as in vertebrates, amphioxus Gbx and the Hox cluster are on the same chromosome. From analysis of linked genes, we argue that during evolution a single ancestral Gbx gene duplicated fourfold in vertebrates, with subsequent loss of two duplicates. Amphioxus Gbx is expressed in all germ layers in the posterior 75% of the embryo, and in the CNS, the Gbx and Otx domains abut at the boundary between the cerebral vesicle (forebrain/midbrain) and the hindbrain. Thus, the genetic machinery to position the MHB was present in the protochordate ancestors of the vertebrates, but is insufficient for induction of organizer genes. Comparison with hemichordates suggests that anterior Otx and posterior Gbx domains were probably overlapping in the ancestral deuterostome and came to abut at the MHB early in the chordate lineage before MHB organizer properties evolved.