Colonial Marine Invertebrates Research Articles

A family of unusual immunoglobulin superfamily genes in an invertebrate histocompatibility complex

Most colonial marine invertebrates are capable of allorecognition, the ability to distinguish between themselves and conspecifics. One long-standing question is whether invertebrate allorecognition genes are homologous to vertebrate histocompatibility genes. In the cnidarian Hydractinia symbiolongicarpus, allorecognition is controlled by at least two genes, Allorecognition 1 (Alr1) and Allorecognition 2 (Alr2), which encode highly polymorphic cell-surface proteins that serve as markers of self. Here, we show that Alr1 and Alr2 are part of a family of 41 Alr genes, all of which reside in a single genomic interval called the Allorecognition Complex (ARC). Using sensitive homology searches and highly accurate structural predictions, we demonstrate that the Alr proteins are members of the immunoglobulin superfamily (IgSF) with V-set and I-set Ig domains unlike any previously identified in animals. Specifically, their primary amino acid sequences lack many of the motifs considered diagnostic for V-set and I-set domains, yet they adopt secondary and tertiary structures nearly identical to canonical Ig domains. Thus, the V-set domain, which played a central role in the evolution of vertebrate adaptive immunity, was present in the last common ancestor of cnidarians and bilaterians. Unexpectedly, several Alr proteins also have immunoreceptor tyrosine-based activation motifs and immunoreceptor tyrosine-based inhibitory motifs in their cytoplasmic tails, suggesting they could participate in pathways homologous to those that regulate immunity in humans and flies. This work expands our definition of the IgSF with the addition of a family of unusual members, several of which play a role in invertebrate histocompatibility.

Fission in a colonial marine invertebrate signifies unique life history strategies rather than being a demographic trait

Each of the few known life-history strategies (e.g., r/K and parity [semelparity and iteroparity]), is a composite stratagem, signified by co-evolved sets of trade-offs with stochastically distributed variations that do not form novel structured strategies. Tracking the demographic traits of 81 Botryllus schlosseri (a marine urochordate) colonies, from birth to death, we revealed three co-existing novel life-history strategies in this long-standing laboratory-bred population, all are bracketed through colonial fission (termed NF, FA and FB for no fission, fission after and fission before reaching maximal colony size, respectively) and derived from organisms maintained in a benign, highly invariable environment. This environment allows us to capture the strategists’ blueprints and their net performance through 13 traits, each branded by high within-strategy variation. Yet, six traits differed significantly among the strategies and, in two, the FB was notably different. These results frame fissions in colonial organisms not as demographic traits, but as pivotal agents for life-history strategies.

When does growth rate influence fitness in a colonial marine invertebrate?

Growth rate affects body size, and larger body sizes are often associated with the capacity to produce more surviving offspring. However, the assumption that growth rate should positively relate to fitness is rarely tested, especially in colonial marine invertebrates where size and age can be decoupled. We measured growth, survival, and reproduction through repeated census of 97 colonies from two populations of a marine bryozoan in the field from settlement to the end of their reproductive season in the northern Gulf of Mexico. Despite large population differences in fitness when grown in a common garden setting, selection within populations on variation in relative growth rate prior to reproduction was similar. In both populations, colonies that grew faster early after settlement, hence were larger, did not consistently have higher fitness than colonies that grew slower after settlement. Instead, early juvenile growth was uncorrelated to later juvenile growth, and colonies that grew faster just prior to the onset of reproduction had higher fitness than colonies that grew slower during this time. Growth rates then declined with the onset of reproduction. Our results show that, rather than being a simple consequence of selection on body size, growth rate can directly affect variation in fitness in ways that are not directly attributable to juvenile size. Colony size and growth rate in modular animals are not always reliable surrogates for direct estimates of survival and reproduction, without identifying when and how growth affects fitness.

Umbellula pomona sp. nov., a new sea pen from Mar del Plata Submarine Canyon (Cnidaria: Octocorallia: Pennatulacea)

Sea pens (Cnidaria: Anthozoa: Pennatulacea) constitute a distinctive group of colonial marine invertebrates. They inhabit the world`s oceans, from shallow to deep waters. Studies about this group in Argentina are scarce, and no species have been described in the area in over a decade. Based on samples collected in Mar del Plata Submarine Canyon at about 3000 m deep we describe a new species of sea pen, Umbellula pomona Risaro, Williams & Lauretta sp. nov. This is a spiculate Umbellula that differs from other species of Umbellula with sclerites, by the number, development and distribution of the autozooids in its terminal cluster, as well as the shape of its axis. Molecular data also distinguishes it from other known species. Of the forty-three described species approximately ten are considered valid for the genus Umbellula, four of them are registered for the South Atlantic Ocean and only three are described for the Antarctic region. Since sampling efforts in this area have been scarce, the number of species of sea pens from the region is likely to increase substantially in the coming years.

An ecological driver for the macroevolution of morphological polymorphism within colonial invertebrates.

Colonial marine invertebrates, such as corals and bryozoans, have modular growth. Individual modules within a colony are homologous to an individual solitary animal body. But in contrast to the predominately sexual origin of solitary animal bodies, modules within a colony are always produced asexually. The repetition of modules and the indeterminism of their organization gives colonies the ability to grow in ways solitary animals cannot. Colonial invertebrates consequently grow in such a way as to resemble weeds, bushes, or trees. The multitude of growth forms of colonial invertebrates arise from differences how individual colonies within a species tend to invest their energy into modular growth, persistence, asexual propagation, and sexual reproduction. Moreover, many colonial invertebrates possess several body types, morphological polymorphism among modules, where modules qualitatively differ in shape, size, and function. In this paper, I propose a mechanism that links the origin of novel body types to the evolution of life‐history strategies among species. When colonies first evolve from solitary ancestors, the life‐history strategy of the colony remains constrained by the life‐history strategies of the individual modules within the colony until a new polymorph type evolves. The addition of novel body types within a colony introduces potential variation in life‐history strategies. Colonies can then change strategies by regulating the frequencies of body types within the colony. This, along with the ability of body types to simplify their structure permits colonies to evolve more complex life‐histories. Each new polymorph type that evolves permits more variation in colonial life‐histories to exist.

Architecture of Anoteropora latirostris (Bryozoa, Cheilostomata) and implications for their biomineralization

Cheilostome Bryozoa Anoteropora latirostris, a colonial marine invertebrate, constructs its skeleton from calcite and aragonite. This study presents firstly correlated multi-scale electron microscopy, micro-computed tomography, electron backscatter diffraction and NanoSIMS mapping. We show that all primary, coarse-grained platy calcitic lateral walls are covered by fine-grained fibrous aragonite. Vertical lateral walls separating autozooid chambers have aragonite only on their distal side. This type of asymmetric mineralization of lateral walls results from the vertical arrangement of the zooids at the growth margins of the colony and represents a type of biomineralization previously unknown in cheilostome bryozoans. NanoSIMS mapping across the aragonite-calcite interface indicates an organic layer between both mineral phases, likely representing an organic template for biomineralization of aragonite on the calcite layer. Analysis of crystallographic orientations show a moderately strong crystallographic preferred orientation (CPO) for calcite (7.4 times random orientation) and an overall weaker CPO for aragonite (2.4 times random orientation) with a high degree of twinning (45%) of the aragonite grains. The calculated Young’s modulus for the CPO map shows a weak mechanical direction perpendicular to the colony’s upper surface facilitating this organism’s strategy of clonal reproduction by fragmentation along the vertical zooid walls.

Invertebrate allorecognition

Most colonial marine invertebrates live as surface encrustations in benthic environments. As they grow, these animals frequently encounter other members of their own species. These encounters typically lead to conflict, in which the colonies aggressively compete for space, or co-existence, in which the colonies peacefully border each other. Sometimes, however, interacting colonies will engage in a form of cooperation in which they fuse together and actively share resources.

Influence of food, body size, and fragmentation on metabolic rate in a sessile marine invertebrate

AbstractMetabolic rates vary among individuals according to food availability and phenotype, most notably body size. Disentangling size from other factors (e.g., age, reproductive status) can be difficult in some groups, but modular organisms may provide an opportunity for manipulating size experimentally. While modular organisms are increasingly used to understand metabolic scaling, the potential of feeding to alter metabolic scaling has not been explored in this group. Here, we perform a series of experiments to examine the drivers of metabolic rate in a modular marine invertebrate, the bryozoan Bugula neritina. We manipulated size and examined metabolic rate in either fed or starved individuals to test for interactions between size manipulation and food availability. Field collected colonies of unknown age showed isometric metabolic scaling, but those colonies in which size was manipulated showed allometric scaling. To further disentangle age effects from size effects, we measured metabolic rate of individuals of known age and again found allometric scaling. Metabolic rate strongly depended on access to food: starvation decreased metabolic rate by 20% and feeding increased metabolic rate by 43%. In comparison to other marine invertebrates, however, the increase in metabolic rate, as well as the duration of the increase (known as specific dynamic action, SDA), were both low. Importantly, neither starvation nor feeding altered the metabolic scaling of our colonies. Overall, we found that field‐collected individuals showed isometric metabolic scaling, whereas metabolic rate of size‐manipulated colonies scaled allometrically with body size. Thus, metabolic scaling is affected by size manipulation but not feeding in this colonial marine invertebrate.

Whole-Body Regeneration in the Colonial Tunicate Botrylloides leachii.

The colonial marine invertebrate Botrylloides leachii belongs to the Tunicata subphylum, the closest invertebrate relatives to the vertebrate group and the only known class of chordates that can undergo whole-body regeneration (WBR). This dramatic developmental process allows a minute isolated fragment of B. leachii's vascular system, or a colony excised of all adults, to restore a functional animal in as little as 10 days. In addition to this exceptional regenerative capacity, B. leachii can reproduce both sexually, through a tadpole larval stage, and asexually, through palleal budding. Thus, three alternative developmental strategies lead to the establishment of filter-feeding adults. Consequently, B. leachii is particularly well suited for comparative studies on regeneration and should provide novel insights into regenerative processes in chordates.Here, after a short introduction on regeneration, we overview the biology of B. leachii as well as the current state of knowledge on WBR in this species and in related species of tunicates. Finally, we highlight the possible future directions that research might take in the study of WBR, including thoughts on technological approaches that appear most promising in this context. Overall, we provide a synthesis of the current knowledge on WBR in B. leachii to support research in this chordate species.

Coupling astogenic aging in the colonial tunicate Botryllus schlosseri with the stress protein mortalin

Botryllus schlosseri, a colonial marine invertebrate, exhibits three generations of short-lived astogenic modules that continuously grow and die throughout the colony's entire lifespan, within week-long repeating budding cycles (blastogenesis), each consisting of four stages (A-D). At stage D, aging is followed by the complete absorption of adult modules (zooids) via a massive apoptotic process. Here we studied in Botryllus the protein mortalin (HSP70s member), a molecule largely known for its association with aging and proliferation. In-situ hybridization and qPCR assays reveal that mortalin follows the cyclic pattern of blastogenesis. Colonies at blastogenic stage D display the highest mortalin levels, and young modules exhibit elevated mortalin levels compared to old modules. Manipulations of mortalin with the specific allosteric inhibitor MKT-077 has led to a decrease in the modules’ growth rate and the development of abnormal somatic/germinal morphologies (primarily in vasculature and in organs such as the endostyle, the stomach and gonads). We therefore propose that mortalin plays a significant role in the astogeny and aging of colonial modules in B. schlosseri, by direct involvement in the regulation of blastogenesis.

Does energy flux predict density-dependence? An empirical field test.

Changes in population density alter the availability, acquisition, and expenditure of resources by individuals, and consequently their contribution to the flux of energy in a system. While both negative and positive density-dependence have been well studied in natural populations, we are yet to estimate the underlying energy flows that generate these patterns and the ambivalent effects of density make prediction difficult. Ultimately, density-dependence should emerge from the effects of conspecifics on rates of energy intake (feeding) and expenditure (metabolism) at the organismal level, thus determining the discretionary energy available for growth. Using a model system of colonial marine invertebrates, we measured feeding and metabolic rates across a range of population densities to calculate how discretionary energy per colony changes with density and test whether this energy predicts observed patterns in organismal size across densities. We found that both feeding and metabolic rates decline with density but that feeding declines faster, and that this discrepancy is the source of density-dependent reductions in individual growth. Importantly, we could predict the size of our focal organisms after eightweeks in the field based on our estimates of energy intake and expenditure. The effects of density on both energy intake and expenditure overwhelmed the effects of body size; even though higher density populations had smaller colonies (with higher mass-specific biological rates), density effects meant that these smaller colonies had lower mass-specific rates overall. Thus, to predict the contribution of organisms to the flux of energy in populations, it seems necessary not only to quantify how rates of energy intake and expenditure scale with body size, but also how they scale with density given that this ecological constraint can be a stronger driver of energy use than the physiological constraint of body size.

Temperature effects on mass-scaling exponents in colonial animals: a manipulative test.

Body size and temperature are fundamental drivers of ecological processes because they determine metabolic rates at the individual level. Whether these drivers act independently on individual-level metabolic rates remains uncertain. Most studies of intraspecific scaling of unitary organisms must rely on preexisting differences in size to examine its relationship with metabolic rate, thereby potentially confounding size-correlated traits (e.g., age, nutrition) with size, which can affect metabolic rate. Here, we use a size manipulation approach to test whether metabolic mass scaling and temperature dependence interact in four species (two phyla) of colonial marine invertebrates. Size manipulation in colonial organisms allows tests of how ecological processes (e.g., predation) affect individual physiology and consequently population- and community-level energy flux. Body mass and temperature interacted in two species, with one species exhibiting decreased and the other increased mass-scaling exponents with increasing temperature. The allometric scaling of metabolic rate that we observe in three species contrasts with the isometric scaling of ingestion rates observed in some colonial marine invertebrates. Thus, we suggest that the often observed competitive superiority of colonial over unitary organisms may arise because the difference between energy intake and expenditure increases more strongly with size in colonial organisms.

Environment-dependent variation in selection on life history across small spatial scales.

Variation in life-history traits is ubiquitous, even though genetic variation is thought to be depleted by selection. One potential mechanism for the maintenance of trait variation is spatially variable selection. We explored spatial variation in selection in the field for a colonial marine invertebrate that shows phenotypic differences across a depth gradient of only 3 m. Our analysis included life-history traits relating to module size, colony growth, and phenology. Directional selection on colony growth varied in strength across depths, while module size was under directional selection at one depth but not the other. Differences in selection may explain some of the observed phenotypic differentiation among depths for one trait but not another: instead, selection should actually erode the differences observed for this trait. Our results suggest selection is not acting alone to maintain trait variation within and across environments in this system.

More than one genotype: how common is intracolonial genetic variability in scleractinian corals?

In recent years, a few colonial marine invertebrates have shown intracolonial genetic variability, a previously unreported phenomenon. Intracolonial genetic variability describes the occurrence of more than a single genotype within an individual colony. This variability can be traced back to two underlying processes: chimerism and mosaicism. Chimerism is the fusion of two or more individuals, whereas mosaicism mostly derives from somatic cell mutations. Until now, it remained unclear to what degree the ecologically important group of hermatypic (reef building) corals might be affected. We investigate the occurrence of intracolonial genetic variability in five scleractinian corals: Acropora florida, Acropora hyacinthus, Acropora sarmentosa, Pocillopora species complex and Porites australiensis. The main focus was to test different genera for the phenomenon via microsatellite markers and to distinguish which underlying process caused the genetic heterogeneity. Our results show that intracolonial genetic variability was common (between 46.6% for A.sarmentosa and 23.8% for P.species complex) in all tested corals. The main process was mosaicism (69 cases of 222 tested colonies), but at least one chimera existed in every species. This suggests that intracolonial genetic variability is widespread in scleractinian corals and could challenge the view of a coral colony as an individual and therefore a unit of selection. However, it might also hold potential for colony survival under rapidly changing environmental conditions.

Friend or foe? How an invertebrate distinguishes self from non-self in a natural histocompatibility interaction. (TRAN3P.912)

Abstract The existence of sophisticated self/non-self systems is not limited to vertebrates. Several colonial marine invertebrates have evolved systems that distinguish between conspecifics via cell-cell contact. This phenomenon, known as allorecognition, occurs when colonies encounter each other as they grow across their substrate. Compatible colonies fuse or co-exist, while incompatible colonies reject and often aggressively compete for space. In all taxa studied to date, allorecognition phenotypes are determined by highly polymorphic loci, which ensure that colonies are only compatible with themselves or close kin. How allorecognition molecules achieve this specificity remains unknown. Allorecognition in Hydractinia is controlled by at least two genes, alr1 and alr2, which encode highly polymorphic transmembrane proteins similar to immunoglobulin superfamily molecules. Colonies with matching alleles at alr1 and alr2 fuse, while colonies with no matching alleles reject. Using in vitro assays with recombinant proteins, we demonstrate that alr1 alleles bind to themselves but not to other alr1 alleles. We suggest that, in vivo, compatibility between colonies is also determined by allele-specific homophilic binding of alr proteins. Since fusion is rare in nature and single populations contain hundreds of unique alr alleles, the Hydractinia allorecognition system appears to be based on a biophysical mechanism with unprecedented allelic diversity and specificity.

Faster is not always better: selection on growth rate fluctuates across life history and environments.

Growth rate is increasingly recognized as a key life-history trait that may affect fitness directly rather than evolve as a by-product of selection on size or age. An ongoing challenge is to explain the abundant levels of phenotypic and genetic variation in growth rates often seen in natural populations, despite what is expected to be consistently strong selection on this trait. Such a paradox suggests limits to how contemporary growth rates evolve. We explored limits arising from variation in selection, based on selection differentials for age-specific growth rates expressed under different ecological conditions. We present results from a field experiment that measured growth rates and reproductive output in wild individuals of a colonial marine invertebrate (Hippopodina iririkiensis), replicated within and across the natural range of succession in its local community. Colony growth rates varied phenotypically throughout this range, but not all such variation was available for selection, nor was it always targeted by selection as expected. While the maintenance of both phenotypic and genetic variation in growth rate is often attributed to costs of growing rapidly, our study highlights the potential for fluctuating selection pressures throughout the life history and across environments to play an important role in this process.

Repair of UV-induced DNA damage in shallow water colonial marine species

Ultraviolet radiation (UVR) may cause significant DNA damages when interacting with shallow water organisms, primarily in the tropics. Here we investigate DNA repair kinetics in two shallow water colonial marine invertebrates, the scleractinian coral Seriatopora hystrix and the colonial tunicate Botryllus schlosseri. Cells from both species were irradiated with UVA/UVB in vitro. Using the alkaline (B. schlosseri) and the neutral (S. hystrix) versions of the comet assay, DNA damage and repair were measured. After 24h, little DNA repair was recorded in the irradiated Botryllus cells for both types of radiation (UVB 3.55W/s/UVA 8.09W/s, 15min radiation each), revealing twice (30%) as much apoptotic cells as controls. Cells of S. hystrix showed a highly efficient DNA repair capacity (i.e. complete DNA repair within 30min post-irradiation), coincident with their endurance to reactive oxygen species impacts. Therefore, while S. hystrix and likely other shallow water corals use efficient DNA repair mechanisms, B. schlosseri and possibly other similar species are reliant on UVR sheltered sites to maintain DNA integrity. The low repair capacity of B. schlosseri might be explained by the ‘disposable soma’ concept, where zooidal soma in this species is cast off and recycled on a weekly basis. Further research is needed to elucidate the spectrum of responses used by shallow water sedentary marine invertebrates to cope with daily high UVR levels.

EVOLUTIONARY CONSTRAINTS AND THE MAINTENANCE OF INDIVIDUAL SPECIALIZATION THROUGHOUT SUCCESSION

Constraints on life-history traits, with their close links to fitness, are widely invoked as limits to niche expansion at most organizational levels. Theoretically, such constraints can maintain individual specialization by preventing adaptation to all niches available, but empirical evidence of them remains elusive for natural populations. This problem may be compounded by a tendency to seek constraints involving multiple traits, neglecting their added potential to manifest in trait expression across environments (i.e., within reaction norms). By replicating genotypes of a colonial marine invertebrate across successional stages in its local community, and taking a holistic approach to the analysis of ensuing reaction norms for fitness, we show the potential for individual specialization to be maintained by genetic constraints associated with these norms, which limit the potential for fitness at one successional stage to improve without loss of fitness at others. Our study provides new insight into the evolutionary maintenance of individual specialization in natural populations and reinforces the importance of reaction norms for studying this phenomenon.

To Fuse or Not to Fuse? An Evolutionary View of Self-Recognition Systems

Self-recognition systems preventing chimera formation following somatic fusion between members of the same species have evolved only in certain phyla (e.g., fungi, cnidarians, poriferans, bryozoans, urochordates). We present here some of the biological features common to fungi and colonial marine invertebrates, which may have driven the evolution of such self-recognition systems. We conclude that the evolution of self-recognition mechanisms in fusible organisms is more likely to result from a complex trade-off between selection pressures linked with a gregariousness and sessile way of life. This trade-off also raises the question of how extrinsic interactions between a group of cooperative cells and its direct environment may have driven the evolution towards a form of individuality, via the emergence self-recognition systems in fusible organisms. Gregariousness and sessility being two features intrinsically associated emergence of multicellularity, somatic incompatibility systems could then be considered as one of the first expressions of individuality in early multicellular organisms.

Marine invertebrates cross phyla comparisons reveal highly conserved immune machinery

Naturally occurring histocompatibility responses, following tissue-to-tissue allogeneic contacts, are common among numerous colonial marine invertebrate taxa, including sponges, cnidarians, bryozoans and ascidians. These responses, often culminating in either tissue fusions or rejections, activate a wide array of innate immune components. By comparing two allorejection EST libraries, developed from alloincompatible challenged colonies of the stony coral Stylophora pistillata and the ascidian Botryllus schlosseri, we revealed a common basis for innate immunity in these two evolutionary distant species. Two prominent genes within this common basis were the immunophilins, Cyclophilin A (CypA) and FK506-binding protein (FKBP). In situ hybridizations revealed that mRNA expression of the coral and ascidian immunophilins was restricted to specific allorecognition effector cell populations (nematoblasts and nematocytes in the coral and morula cells in the ascidian). The expressions were limited to only some of the effector cells within a population, disclosing disparities in numbers and location between naïve colonies and their immune challenged counterparts. Administration of the immunosuppression drug Cyclosporine-A during ascidian's allogeneic assays inhibited both fusion and rejection reactions, probably through the inhibition of ascidian's immunocytes (morula cells) movement and activation. Our results, together with previous published data, depict an immunophilins-based immune mechanism, which is similarly activated in allogeneic responses of distantly related animals from sponges to humans.