Seasonal tissue-specific gene expression in wild crown-of-thorns starfish reveals reproductive and stress-related transcriptional systems.

Animals are influenced by the season, yet we know little about the changes that occur in most species throughout the year. This is particularly true in tropical marine animals that experience relatively small annual temperature and daylight changes. Like many coral reef inhabitants, the crown-of-thorns starfish (COTS), well known as a notorious consumer of corals and destroyer of coral reefs, reproduces exclusively in the summer. By comparing gene expression in 7 somatic tissues procured from wild COTS sampled on the Great Barrier Reef, we identified more than 2,000 protein-coding genes that change significantly between summer and winter. COTS genes that appear to mediate conspecific communication, including both signalling factors released into the surrounding sea water and cell surface receptors, are up-regulated in external secretory and sensory tissues in the summer, often in a sex-specific manner. Sexually dimorphic gene expression appears to be underpinned by sex- and season-specific transcription factors (TFs) and gene regulatory programs. There are over 100 TFs that are seasonally expressed, 87% of which are significantly up-regulated in the summer. Six nuclear receptors are up-regulated in all tissues in the summer, suggesting that systemic seasonal changes are hormonally controlled, as in vertebrates. Unexpectedly, there is a suite of stress-related chaperone proteins and TFs, including HIFa, ATF3, C/EBP, CREB, and NF-κB, that are uniquely and widely co-expressed in gravid females. The up-regulation of these stress proteins in the summer suggests the demands of oogenesis in this highly fecund starfish affects protein stability and turnover in somatic cells. Together, these circannual changes in gene expression provide novel insights into seasonal changes in this coral reef pest and have the potential to identify vulnerabilities for targeted biocontrol.

Protecting Great Barrier Reef resilience through effective management of crown-of-thorns starfish outbreaks.

Resilience-based management is essential to protect ecosystems in the Anthropocene. Unlike large-scale climate threats to Great Barrier Reef (GBR) corals, outbreaks of coral-eating crown-of-thorns starfish (COTS; Acanthaster cf. solaris) can be directly managed through targeted culling. Here, we evaluate the outcomes of a decade of strategic COTS management in suppressing outbreaks and protecting corals during the 4th COTS outbreak wave at reef and regional scales (sectors). We compare COTS density and coral cover dynamics during the 3rd and 4th outbreak waves. During the 4th outbreak wave, sectors that received limited to no culling had sustained COTS outbreaks causing significant coral losses. In contrast, in sectors that received timely and sufficient cull effort, coral cover increased substantially, and outbreaks were suppressed with COTS densities up to six-fold lower than in the 3rd outbreak wave. In the Townsville sector for example, despite exposure to comparable disturbance regimes during the 4th outbreak wave, effective outbreak suppression coincided with relative increases in sector-wide coral cover (44%), versus significant coral cover declines (37%) during the 3rd outbreak wave. Importantly, these estimated increases span entire sectors, not just reefs with active COTS control. Outbreaking reefs with higher levels of culling had net increases in coral cover, while the rate of coral loss was more than halved on reefs with lower levels of cull effort. Our results also indicate that outbreak wave progression to adjoining sectors has been delayed, probably via suppression of COTS larval supply. Our findings provide compelling evidence that proactive, targeted, and sustained COTS management can effectively suppress COTS outbreaks and deliver coral growth and recovery benefits at reef and sector-wide scales. The clear coral protection outcomes demonstrate the value of targeted manual culling as both a scalable intervention to mitigate COTS outbreaks, and a potent resilience-based management tool to "buy time" for coral reefs, protecting reef ecosystem functions and biodiversity as the climate changes.

Crown-of-thorns seastar (Acanthaster spp.) feeding ecology across species and regions.

The coral predators, crown-of-thorns starfish (COTS, Acanthaster spp.) remain a major cause of extensive and widespread coral loss in Indo-Pacific coral reefs. With increased phylogenetic understanding of these seastars, at least five species appear to be present across different regions. We compare the feeding ecology of these species. Where acroporid corals are prevalent, Acanthaster spp. often exhibit a preference for these corals, with Porites being least preferred, as seen in most species including Acanthaster planci in the northern Indian Ocean and Acanthaster cf. solaris in the west Pacific. In the eastern Pacific, where Acropora is largely absent, Acanthaster cf. ellisii prey on a range of coral species, including Porites. Coral predation by COTS is influenced by several factors including food availability, coral nutritional value, protective crustaceans and coral defenses, with differences in feeding ecology and behaviour emerging across the different COTS species. Feeding behaviour of COTS can act to increase coral species richness by reducing the dominance of fast-growing species. In outbreaking populations, COTS impacts reef systems by reducing live coral cover, eroding reef complexity and causing shifts in reef trophic structure. Where data are available, we synthesise and contrast the feeding preferences and foraging behaviour of Acanthaster species, and their impact on coral assemblages across the different species and regions. For areas where focal predation on Acropora occurs, also the fastest growing coral with the greatest recovery potential following mass mortality events, the combination of climate change and COTS outbreaks presents an imminent threat to coral reefs. This is exacerbated by the dietary flexibility of Acanthaster species. The impacts of heatwaves, COTS and other stressors are creating a negative feedback loop accelerating coral reef decline.

A description of the bat star nervous system throughout larval ontogeny.

Larvae represent a distinct life history stage in which animal morphology and behavior contrast strongly to adult organisms. This life history stage is a ubiquitous aspect of animal life cycles, particularly in the marine environment. In many species, the structure and function of the nervous system differ significantly between metamorphosed juveniles and larvae. However, the distribution and diversity of neural cell types in larval nervous systems remains incompletely known. Here, the expression of neurotransmitter and neuropeptide synthesis and transport genes in the bat star Patiria miniata is examined throughout larval development. This characterization of nervous system structure reveals three main neural regions with distinct but overlapping territories. These regions include a densely innervated anterior region, an enteric neural plexus, and neurons associated with the ciliary band. In the ciliary band, cholinergic cells are pervasive while dopaminergic, noradrenergic, and GABAergic cells show regional differences in their localization patterns. Furthermore, the distribution of some neural subtypes changes throughout larval development, suggesting that changes in nervous system structure align with shifting ecological priorities during different larval stages, before the development of the adult nervous system. While past work has described aspects of P. miniata larval nervous system structure, largely focusing on early developmental timepoints, this work provides a comprehensive description of neural cell type localization throughout the extensive larval period.

Improving coral cover using an integrated pest management framework.

Integrated pest management (IPM) leverages our understanding of ecological interactions to mitigate the impact of pest species on economically and/or ecologically important assets. It has primarily been applied in terrestrial settings (e.g., agriculture), but has rarely been attempted for marine ecosystems. The crown-of-thorns starfish (CoTS), Acanthaster spp., is a voracious coral predator throughout the Indo-Pacific where it undergoes large population increases (irruptions), termed outbreaks. During outbreaks CoTS act as a pest species and can result in substantial coral loss. Contemporary management of CoTS on the Great Barrier Reef (GBR) adopts facets of the IPM paradigm to manage these outbreaks through strategic use of direct manual control (culling) of individuals in response to ecologically based target thresholds. There has, however, been limited quantitative analysis of how to optimize the implementation of such thresholds. Here we use a multispecies modeling approach to assess the performance of alternative CoTS management scenarios for improving coral cover trajectories. The scenarios examined varied in terms of their ecological threshold target, the sensitivity of the threshold, and the level of management resourcing. Our approach illustrates how to quantify multidimensional trade-offs in resourcing constraints, concurrent CoTS and coral population dynamics, the stringency of target thresholds, and the geographical scale of management outcomes (number of sites). We found strategies with low target density thresholds for CoTS (≤0.03 CoTS min-1 ) could act as "Effort Sinks" and limit the number of sites that could be effectively controlled, particularly under CoTS population outbreaks. This was because a handful of sites took longer to control, which meant other sites were not controlled. Higher density thresholds (e.g., 0.04-0.08 CoTS min-1 ), tuned to levels of coral cover, diluted resources among sites but were more robust to resourcing constraints and pest population dynamics. Our study highlights trade-off decisions when using an IPM framework and informs the implementation of threshold-based strategies on the GBR.

Regeneration of starfish radial nerve cord restores animal mobility and unveils a new coelomocyte population.

The potential to regenerate a damaged body part is expressed to a different extent in animals. Echinoderms, in particular starfish, are known for their outstanding regenerating potential. Differently, humans have restricted abilities to restore organ systems being dependent on limited sources of stem cells. In particular, the potential to regenerate the central nervous system is extremely limited, explaining the lack of natural mechanisms that could overcome the development of neurodegenerative diseases and the occurrence of trauma. Therefore, understanding the molecular and cellular mechanisms of regeneration in starfish could help the development of new therapeutic approaches in humans. In this study, we tackle the problem of starfish central nervous system regeneration by examining the external and internal anatomical and behavioral traits, the dynamics of coelomocyte populations, and neuronal tissue architecture after radial nerve cord (RNC) partial ablation. We noticed that the removal of part of RNC generated several anatomic anomalies and induced behavioral modifications (injured arm could not be used anymore to lead the starfish movement). Those alterations seem to be related to defense mechanisms and protection of the wound. In particular, histology showed that tissue patterns during regeneration resemble those described in holothurians and in starfish arm tip regeneration. Flow cytometry coupled with imaging flow cytometry unveiled a new coelomocyte population during the late phase of the regeneration process. Morphotypes of these and previously characterized coelomocyte populations were described based on IFC data. Further studies of this new coelomocyte population might provide insights on their involvement in radial nerve cord regeneration.

Sea stars resist wasting through active immune and collagen systems.

Epidemics are becoming more common and severe, however, pinpointing the causes can be challenging, particularly in marine environments. The cause of sea star wasting (SSW) disease, the ongoing, largest known panzootic of marine wildlife, is unresolved. Here, we measured gene expression longitudinally of 24 adult Pisaster ochraceus sea stars, collected from a recovered site, as they remained asymptomatic (8 individuals) or naturally progressed through SSW (16 individuals) in individual aquaria. Immune, tissue integrity and pro-collagen genes were more highly expressed in asymptomatic relative to wasting individuals, while hypoxia-inducible factor 1-α and RNA processing genes were more highly expressed in wasting relative to asymptomatic individuals. Integrating microbiome data from the same tissue samples, we identified genes and microbes whose abundance/growth was associated with disease status. Importantly, sea stars that remained visibly healthy showed that laboratory conditions had little effect on microbiome composition. Lastly, considering genotypes at 98 145 single-nucleotide polymorphism, we found no variants associated with final health status. These findings suggest that animals exposed to the cause(s) of SSW remain asymptomatic with an active immune response and sustained control of their collagen system while animals that succumb to wasting show evidence of responding to hypoxia and dysregulation of RNA processing systems.

The C-terminally amidated relaxin-like gonad-stimulating peptide in the starfish Astropecten scoparius.

A relaxin-like gonad-stimulating peptide (RGP) in starfish was the first identified invertebrate gonadotropin, consisting of A- and B-chain. Recently, an RGP ortholog (Asc-RGP) from Astropecten scoparius in the order Paxillosida was found to harbor an amidation signal (Gly-Arg) at the C-terminus of the B-chain (Mita et al., 2020a). Two cleavage sites were also predicted within the signal peptide of the Asc-RGP precursor. Thus, four kinds of analogs (Asc-RGP-NH2(S), Asc-RGP-GR(S), Asc-RGP- NH2(L), Asc-RGP-GR(L) were hypothesized as natural Asc-RGPs. To identify the natural Asc-RGP, an extract of radial nerve cords from A. scoparius was analyzed using reverse-phase high-performance liquid chromatography and MALDI-TOF-mass spectrometry. The molecular weight of Asc-RGP was 4585.3, and those of A- and B-chains were 2511.8 and 2079.8, respectively. This strongly suggests that natural RGP in A. scoparius is Asc-RGP-NH2(S). Asc-RGP-NH2(S) stimulated 1-methyladenine and cyclic AMP production in isolated ovarian follicle cells of A. scoparius. On the other hand, the concentrations of four synthetic Asc-RGP analogs required for the induction of spawning in 50% of ovarian fragments were almost the same. The size and C-terminal amidation of the B-chain might not be important for spawning-inducing activity. C-terminally amidated RGPs in the B-chain were also observed in other species of starfish belonging to the order Paxillosida, particularly the family Astropectinidae, but not the family Luidiidae.

Biochemical metabolomic profiling of the Crown-of-Thorns Starfish (Acanthaster): New insight into its biology for improved pest management.

The Crown-of-Thorns Starfish (COTS), Acanthaster species, is a voracious coral predator that destroys coral reefs when in outbreak status. The baseline metabolite and lipid biomolecules of 10 COTS tissues, including eggs from gravid females, were investigated in this study to provide insight into their biology and identify avenues for control. Targeted and untargeted metabolite- and lipidomics-based mass spectrometry approaches were used to obtain tissue-specific metabolite and lipid profiles. Across all COTS tissues, 410 metabolites and 367 lipids were identified. Most abundant were amino acids and peptides (18.7%) and wax esters (17%). There were 262 metabolites and 192 lipids identified in COTS eggs. Wax esters were more abundant in the eggs (30%) followed by triacylglycerols (TG), amino acids, and peptides. The diversity of asterosaponins in eggs (34) was higher than in tissues (2). Several asterosaponins known to modulate sperm acrosome reaction were putatively identified, including glycoside B, asterosaponin-4 (Co-Aris III), and regularoside B (asterosaponin A). The saponins saponin A, thornasteroside A, hillaside B, and non-saponins dictyol J and axinellamine B which have been shown to possess defensive properties, were found in abundance in gonads, skin, and radial nerve tissues. Inosine and 2-hexyldecanoic acid are the most abundant metabolites in tissues and eggs. As a secondary metabolite of purine degradation, inosine plays an important role in purine biosynthesis, while 2-hexyldecanoic acid is known to suppress side-chain crystallization during the synthesis of amphiphilic macromolecules (i.e., phospholipids). These significant spatial changes in metabolite, lipid, and asterosaponin profiles enabled unique insights into key biological tissue-specific processes that could be manipulated to better control COTS populations. Our findings highlight COTS as a novel source of molecules with therapeutic and cosmetic properties (ceramides, sphingolipids, carnosine, and inosine). These outcomes will be highly relevant for the development of strategies for COTS management including chemotaxis-based biocontrol and exploitation of COTS carcasses for the extraction of commercial molecules.

Regeneration of the larval sea star nervous system by wounding induced respecification to the Sox2 lineage.

The ability to restore lost body parts following traumatic injury is a fascinating area of biology that challenges current understanding of the ontogeny of differentiation. The origin of new cells needed to regenerate lost tissue, and whether they are pluripotent or have de- or trans-differentiated, remains one of the most important open questions . Additionally, it is not known whether developmental gene regulatory networks are reused or whether regeneration specific networks are deployed. Echinoderms, including sea stars, have extensive ability for regeneration, however, the technologies for obtaining transgenic echinoderms are limited and tracking cells involved in regeneration, and thus identifying the cellular sources and potencies has proven challenging. In this study, we develop new transgenic tools to follow the fate of populations of cells in the regenerating larva of the sea star Patiria miniata. We show that the larval serotonergic nervous system can regenerate following decapitation. Using a BAC-transgenesis approach we show that expression of the pan ectodermal marker, sox2, is induced in previously sox2 minus cells , even when cell division is inhibited. sox2+ cells give rise to new sox4+ neural precursors that then proceed along an embryonic neurogenesis pathway to reform the anterior nervous systems. sox2+ cells contribute to only neural and ectoderm lineages, indicating that these progenitors maintain their normal, embryonic lineage restriction. This indicates that sea star larval regeneration uses a combination of existing lineage restricted stem cells, as well as respecification of cells into neural lineages, and at least partial reuse of developmental GRNs to regenerate their nervous system.

Topological braiding and virtual particles on the cell membrane.

Braiding of topological structures in complex matter fields provides a robust framework for encoding and processing information, and it has been extensively studied in the context of topological quantum computation. In living systems, topological defects are crucial for the localization and organization of biochemical signaling waves, but their braiding dynamics remain unexplored. Here, we show that the spiral wave cores, which organize the Rho-GTP protein signaling dynamics and force generation on the membrane of starfish egg cells, undergo spontaneous braiding dynamics. Experimentally measured world line braiding exponents and topological entropy correlate with cellular activity and agree with predictions from a generic field theory. Our analysis further reveals the creation and annihilation of virtual quasi-particle excitations during defect scattering events, suggesting phenomenological parallels between quantum and living matter.

Divalent metal transporter-related protein restricts animals to marine habitats.

Utilization and regulation of metals from seawater by marine organisms are important physiological processes. To better understand metal regulation, we searched the crown-of-thorns starfish genome for the divalent metal transporter (DMT) gene, a membrane protein responsible for uptake of divalent cations. We found two DMT-like sequences. One is an ortholog of vertebrate DMT, but the other is an unknown protein, which we named DMT-related protein (DMTRP). Functional analysis using a yeast expression system demonstrated that DMT transports various metals, like known DMTs, but DMTRP does not. In contrast, DMTRP reduced the intracellular concentration of some metals, especially zinc, suggesting its involvement in negative regulation of metal uptake. Phylogenetic distribution of the DMTRP gene in various metazoans, including sponges, protostomes, and deuterostomes, indicates that it originated early in metazoan evolution. However, the DMTRP gene is only retained in marine species, and its loss seems to have occurred independently in ecdysozoan and vertebrate lineages from which major freshwater and land animals appeared. DMTRP may be an evolutionary and ecological limitation, restricting organisms that possess it to marine habitats, whereas its loss may have allowed other organisms to invade freshwater and terrestrial habitats.

Sea stars generate downforce to stay attached to surfaces.

Intertidal sea stars often function in environments with extreme hydrodynamic loads that can compromise their ability to remain attached to surfaces. While behavioral responses such as burrowing into sand or sheltering in rock crevices can help minimize hydrodynamic loads, previous work shows that sea stars also alter body shape in response to flow conditions. This morphological plasticity suggests that sea star body shape may play an important hydrodynamic role. In this study, we measured the fluid forces acting on surface-mounted sea star and spherical dome models in water channel tests. All sea star models created downforce, i.e., the fluid pushed the body towards the surface. In contrast, the spherical dome generated lift. We also used Particle Image Velocimetry (PIV) to measure the midplane flow field around the models. Control volume analyses based on the PIV data show that downforce arises because the sea star bodies serve as ramps that divert fluid away from the surface. These observations are further rationalized using force predictions and flow visualizations from numerical simulations. The discovery of downforce generation could explain why sea stars are shaped as they are: the pentaradial geometry aids attachment to surfaces in the presence of high hydrodynamic loads.

Metabolic Responses of Pacific Crown-of-Thorns Sea Stars (Acanthaster sp.) to Acute Warming.

AbstractClimate change and population irruptions of crown-of-thorns sea stars (Acanthaster sp.) are two of the most pervasive threats to coral reefs. Yet there has been little consideration regarding the synergies between ocean warming and the coral-feeding sub-adult and adult stages of this asteroid. Here we explored the thermosensitivity of the aforementioned life stages by assessing physiological responses to acute warming. Thermal sensitivity was assessed based on the maximal activity of enzymes involved in aerobic (citrate synthase) and anaerobic (lactate dehydrogenase) metabolic pathways, as well as the standard metabolic rate of sub-adult and adult sea stars. In both life stages, citrate synthase activity declined with increasing temperature from 15 °C to 40 °C, with negligible activity occurring >35 °C. On the other hand, lactate dehydrogenase activity increased with temperature from 20 °C to 45 °C, indicating a greater reliance on anaerobic metabolism in a warmer environment. The standard metabolic rate of sub-adult sea stars increased with temperature throughout the testing range (24 °C to 36 °C). Adult sea stars exhibited evidence of thermal stress, with metabolic depression occurring from 33 °C. Here, we demonstrate that crown-of-thorns sea stars are sensitive to warming but that adults, and especially sub-adults, may have some resilience to short-term marine heatwaves in the near future.

Optimal Foraging Theory Explains Feeding Preferences in the Western Pacific Crown-of-Thorns Sea Star Acanthaster sp.

AbstractThe selectivity of crown-of-thorns sea stars (Acanthaster sp.) for different coral prey types was quantified in the field and laboratory and compared with a range of nutritional and food quality parameters as well as the growth performance of sea stars fed on different types of coral. Growth rates of small juvenile Acanthaster sp. without previous exposure to coral fed for 6.6 months on 15 individual species of corals showed that the highest rates of growth were achieved on the same types of corals for which adult sea stars show the strongest preference, both in the field and in controlled aquarium conditions. Small Acanthaster sp. (ca. 20 mm, 0.5 g) fed on Acropora formosa, Stylophora pistillata, Seriatopora hystrix, and Pocillopora damicornis increased in size by an average of 9.2-10.7 mm (4.2-5.6 g) per month, compared with 0.1-0.4 mm (0.004-0.028 g) per month on coralline algae fed controls and species such as Porites lutea, Porites lichen, Lobophyllia hemprichii, and Turbinaria mesenterina. Field studies on the same reef where the parents of these juvenile sea stars were collected demonstrated a strong sequential preference for acroporid and then pocilloporid corals, with faviid, merulinid, and poritid corals selected significantly less frequently than other corals when their relative abundance was taken into account. This order of preference by adult field-collected sea stars was confirmed and exhibited even more emphatically in aquarium experiments, where the relative abundance of prey species could be controlled. The growth experiments and measurements of comparative food value between preferred and non-preferred coral prey suggest that feeding preferences in Acanthaster sp. for Acropora and pocilloporids arose consistent with optimal foraging theory and evolved in response to this species being able to feed successfully and efficiently. The high abundance and, therefore, encounter rate of Acropora and pocilloporids is not considered to be an important factor in the evolution of feeding preferences, although relative abundance of alternative prey does affect selectivity. Individual growth and population fitness and reproductive output of Acanthaster sp. will be enhanced by preferential feeding on acroporid and pocilloporid corals, reinforcing the importance of optimal foraging theory in the evolution of feeding preferences.

Identification of SH2 Domain-Mediated Protein Interactions that Operate at Fertilization in the Sea Star Patiria miniata.

The signaling mechanisms controlling internal calcium release at fertilization in animals are still largely unknown. Echinoderms, such as the sea star Patiria miniata, produce abundant and easily accessible sperm and eggs. In addition, eggs are naturally synchronized at the same cell cycle stage, collectively making these animals an attractive model to study the signaling proteins controlling fertilization. However, the lack of antibodies to identify proteins in this model system has slowed progress in identifying key signaling molecules. With the advances in mass spectrometry, we present a method for identifying tyrosine phosphorylated proteins binding to GST-tagged SH2 domains in sea star cell lysates for downstream mass spectrometry analysis.

Recovery of Sea Star Egg Cell Surface Proteins Released at Fertilization.

To provide a better understanding of the composition of the egg cell membrane, we describe a method in which proteins and peptides that are either naturally released by the egg or cleaved by sperm proteases can be collected, analyzed, and identified. Such molecules are captured and isolated from the surrounding seawater via biotinylation, before being concentrated by an affinity interaction and subsequently analyzed by western blotting and mass spectrometry.

Homing behaviour by destructive crown-of-thorns starfish is triggered by local availability of coral prey.

Corallivorous crown-of-thorns starfishes (Acanthaster spp.) can decimate coral assemblages on Indo-Pacific coral reefs during population outbreaks. While initial drivers of population irruptions leading to outbreaks remain largely unknown, subsequent dispersal of outbreaks appears coincident with depletion of coral prey. Here, we used in situ time-lapse photography to characterize movement of the Pacific crown-of-thorns starfish (Acanthaster cf. solaris) in the northern and southern Great Barrier Reef in 2015, during the fourth recorded population outbreak of the starfish, but prior to widespread coral bleaching. Daily tracking of 58 individuals over a total of 1117 h revealed all starfish to move a minimum of 0.52 m, with around half of all tracked starfish showing negligible daily displacement (less than 1 m day-1), ranging up to a maximum of 19 m day-1. Movement was primarily nocturnal and daily displacement varied spatially with variation in local availability of Acropora spp., which is the preferred coral prey. Two distinct behavioural modes emerged: (i) homing movement, whereby tracked paths (as tested against a random-walk-model) involved short displacement distances following distinct 'outward' movement to Acropora prey (typically displaying 'feeding scars') and 'homebound' movement to nearby shelter; versus (ii) roaming movement, whereby individuals showed directional movement beyond initial tracking positions without return. Logistic modelling revealed more than half of all tracked starfish demonstrated homing when local abundance (percentage cover) of preferred Acropora coral prey was greater than 33%. Our results reveal facultative homing by Acanthaster with the prey-dependent behavioural switch to roaming forays providing a mechanism explaining localized aggregations and diffusion of these population irruptions as prey is locally depleted.

A novel G protein-coupled receptor for starfish gonadotropic hormone, relaxin-like gonad-stimulating peptide.

Gonadotropic hormones play important regulatory roles in reproduction. Relaxin-like gonad-stimulating peptide (RGP) is a gonadotropin-like hormone in starfish. However, a receptor for RGP remains to be identified. Here, we describe the identification of an authentic receptor for RGP (RGPR) in the starfish, Patiria pectinifera. A binding assay using radioiodinated P. pectinifera RGP (PpeRGP) revealed that RGPR was expressed in ovarian follicle cells. A RGPR candidate was identified by homology-searching of transcriptome data of P. pectinifera follicle cells. Based on the contig sequences, a putative 947-amino acid PpeRGPR was cloned from follicle cells. Like the vertebrate relaxin family peptide receptors (RXFP 1 and 2), PpeRGPR was a G protein-coupled receptor that harbored a low-density lipoprotein-receptor class A motif and leucine-rich repeat sequences in the extracellular domain of the N-terminal region. Sf9 cells transfected with Gαq16-fused PpeRGPR activated calcium ion mobilization in response to PpeRGP, but not to RGP of another starfish Asterias amurensis, in a dose-dependent fashion. These results confirmed the species-specific reactivity of RGP and the cognate receptor. Thus, the present study provides evidence that PpeRGPR is a specific receptor for PpeRGP. To the best of our knowledge, this is the first report on the identification of a receptor for echinoderm RGP.

A ubiquitous subcuticular bacterial symbiont of a coral predator, the crown-of-thorns starfish, in the Indo-Pacific.

BACKGROUND: Population outbreaks of the crown-of-thorns starfish (Acanthaster planci sensu lato; COTS), a primary predator of reef-building corals in the Indo-Pacific Ocean, are a major threat to coral reefs. While biological and ecological knowledge of COTS has been accumulating since the 1960s, little is known about its associated bacteria. The aim of this study was to provide fundamental information on the dominant COTS-associated bacteria through a multifaceted molecular approach. METHODS: A total of 205 COTS individuals from 17 locations throughout the Indo-Pacific Ocean were examined for the presence of COTS-associated bacteria. We conducted 16S rRNA metabarcoding of COTS to determine the bacterial profiles of different parts of the body and generated a full-length 16S rRNA gene sequence from a single dominant bacterium, which we designated COTS27. We performed phylogenetic analysis to determine the taxonomy, screening of COTS27 across the Indo-Pacific, FISH to visualize it within the COTS tissues, and reconstruction of the bacterial genome from the hologenome sequence data. RESULTS: We discovered that a single bacterium exists at high densities in the subcuticular space in COTS forming a biofilm-like structure between the cuticle and the epidermis. COTS27 belongs to a clade that presumably represents a distinct order (so-called marine spirochetes) in the phylum Spirochaetes and is universally present in COTS throughout the Indo-Pacific Ocean. The reconstructed genome of COTS27 includes some genetic traits that are probably linked to adaptation to marine environments and evolution as an extracellular endosymbiont in subcuticular spaces. CONCLUSIONS: COTS27 can be found in three allopatric COTS species, ranging from the northern Red Sea to the Pacific, implying that the symbiotic relationship arose before the speciation events (approximately 2 million years ago). The universal association of COTS27 with COTS and nearly mono-specific association at least with the Indo-Pacific COTS provides a useful model system for studying symbiont-host interactions in marine invertebrates and may have applications for coral reef conservation. Video Abstract.