Gene regulation of adult skeletogenesis in starfish and modifications during gene network co-option.

The larval skeleton of the echinoderm is believed to have been acquired through co-option of a pre-existing gene regulatory network (GRN); that is, the mechanism for adult skeleton formation in the echinoderm was deployed in early embryogenesis during echinoderm diversification. To explore the evolutionary changes that occurred during co-option, we examined the mechanism for adult skeletogenesis using the starfish Patiria pectinifera. Expression patterns of skeletogenesis-related genes (vegf, vegfr, ets1/2, erg, alx1, ca1, and clect) suggest that adult skeletogenic cells develop from the posterior coelom after the start of feeding. Treatment with inhibitors and gene knockout using transcription activator-like effector nucleases (TALENs) suggest that the feeding-nutrient sensing pathway activates Vegf signaling via target of rapamycin (TOR) activity, leading to the activation of skeletogenic regulatory genes in starfish. In the larval skeletogenesis of sea urchins, the homeobox gene pmar1 activates skeletogenic regulatory genes, but in starfish, localized expression of the pmar1-related genes phbA and phbB was not detected during the adult skeleton formation stage. Based on these data, we provide a model for the adult skeletogenic GRN in the echinoderm and propose that the upstream regulatory system changed from the feeding-TOR-Vegf pathway to a homeobox gene-system during co-option of the skeletogenic GRN.

The Use of Larval Sea Stars and Sea Urchins in the Discovery of Shared Mechanisms of Metazoan Whole-Body Regeneration.

The ability to regenerate is scattered among the metazoan tree of life. Further still, regenerative capacity varies widely within these specific organisms. Numerous organisms, all with different regenerative capabilities, have been studied at length and key similarities and disparities in how regeneration occurs have been identified. In order to get a better grasp on understanding regeneration as a whole, we must search for new models that are capable of extensive regeneration, as well as those that have been under sampled in the literature. As invertebrate deuterostomes, echinoderms fit both of these requirements. Multiple members regenerate various tissue types at all life stages, including examples of whole-body regeneration. Interrogations in two highly studied echinoderms, the sea urchin and the sea star, have provided knowledge of tissue and whole-body regeneration at various life stages. Work has begun to examine regeneration in echinoderm larvae, a potential new system for understanding regenerative mechanisms in a basal deuterostome. Here, we review the ways these two animals' larvae have been utilized as a model of regeneration.

Locomotion and righting behavior of sea stars: a study case on the bat star Asterina stellifera (Asterinidae) / Comportamiento de locomoción y enderezamiento en estrellas de mar: un caso de estudio sobre la estrella Asterina stellifera (Asterinidae)

Introduction: The locomotion behavior of an organism involves the integration of aspects like body symmetry, sensory and locomotor systems. Furthermore, various ecological factors seem to be related to locomotion characteristics, such as foraging strategy, migration trends, response to predators and competitors, and environmental stress. Objective: To analyze locomotion and the influence of body symmetry in the crawling and righting movements of the sea star Asterina stellifera. Methods: We carried out laboratory experiments in aquariums in the presence/absence of water current and on a horizontal and vertical surface. Results: The speed is similar to speed in other species of similar size. Both the speed and linearity of displacement were independent of individual body size. A water current leads to faster crawling and straight paths, but there is no rheotaxis: streams do not affect locomotion. Speed and linearity of displacement were independent of individual body size. The displacement pattern described here may be an adaptation of organisms that present dense populations in communities with high prey abundance, as is the case of A. stellifera. Conclusions: Like other asteroids, this species did not show an Anterior/Posterior plane of symmetry during locomotion, or righting movement: it does not tend to bilaterality.

Introducción: El comportamiento de locomoción de un organismo implica la integración de aspectos como la simetría corporal, los sistemas sensorial y locomotor. Además, varios factores ecológicos parecen estar relacionados con las características de la locomoción, como la estrategia de alimentación, las tendencias migratorias, la respuesta a los depredadores y competidores y el estrés ambiental. Objetivo: Analizar el patrón general de locomoción y la influencia de la simetría corporal en la locomoción y enderezamiento de la estrella de mar Asterina stellifera. Métodos: Realizamos experimentos de laboratorio en acuarios en presencia / ausencia de corriente de agua y en superficie horizontal y vertical. Resultados: La velocidad es similar a la velocidad en otras especies de tamaño similar. Tanto la velocidad como la linealidad del desplazamiento fueron independientes del tamaño corporal individual. Una corriente de agua conduce a una velocidad de desplazamiento mayor y a trayectorias más rectas, pero no hay reotaxis: una corriente de agua no afecta el patrón de locomoción. La velocidad y la linealidad del desplazamiento fueron independientes del tamaño corporal individual. El patrón de desplazamiento aquí descrito puede ser una adaptación de organismos que presentan densas poblaciones en comunidades con alta abundancia de presas, como es el caso de A. stellifera. Conclusiones: Al igual que otros asteroides, esta especie no mostró un plano de simetría Anterior / Posterior durante la locomoción o el movimiento de enderezamiento: no tiende a la bilateralidad.

Variabilidad en el tamaño y alimentación de Acanthaster planci (Echinodermata: Asteroidea) en el sur del Golfo de California, México / Variability of size and food type of Acanthaster planci (Echinodermata: Asteroidea) in the southern Gulf of California, Mexico

Introducción: Es poco lo que se conoce sobre aspectos biológicos y poblacionales de Acanthaster planci en México. Su tamaño se ha relacionado con la cantidad de alimento que consume, por lo que es relevante conocer esta variable. Objetivo: Describir la variabilidad en el diámetro del disco de la especie y de su alimentación en cuatro sitios al sur del Golfo de California. Métodos: De febrero 2008 a marzo 2009 se visitaron mensualmente los sitios. Se registró el diámetro del disco de un total de 389 individuos observados. El diámetro del disco se midió con una cinta métrica flexible colocada de lado a lado, en el borde donde comienzan los brazos. Para facilitar la descripción, el diámetro del disco se clasificó en tres grupos: pequeños ( 26 cm). Del total, 298 organismos se encontraron alimentándose. Se clasificaron los tipos de organismos consumidos en corales duros, otros invertebrados y algas. Para calcular la probabilidad de que individuos del mismo tamaño se alimenten de diferente variedad de organismos, se aplicó el índice de Diversidad de Simpson. Con el fin de detectar diferencias del diámetro del disco entre los meses de estudio y los sitios de muestreo, se aplicó un análisis permutacional de varianza. Resultados: Acanthaster planci tuvo un promedio de diámetro del disco de 18.23 ± 0.21 cm, con un mínimo de 7 cm y un máximo de 36 cm. La estructura de tallas fue unimodal y se caracterizó por la predominancia de individuos medianos, seguida por los pequeños y en baja proporción por los grandes. El diámetro del disco fue estadísticamente más grande en febrero y más pequeño en los últimos meses del año. En San Rafaelito se encontraron los ejemplares más grandes y en Ensenada de Muertos los más pequeños. Los individuos de tamaño mediano fueron más frecuentes a lo largo del año y en todos los sitios. Los individuos pequeños se alimentaron significativamente más de Porites panamensis, seguido de algas coralinas y tapetes. Los más grandes tuvieron los valores de diversidad más bajos, más del 90 % de los individuos en este grupo se alimentaron de Pocillopora spp. y algunos de macroalgas. Los medianos presentaron los valores más altos de diversidad y consumieron todos los tipos de alimento encontrados, sin embargo, como era de esperarse, muchos fueron selectivos hacia Pocillopora spp. Conclusiones: Acanthaster planci se considera un individuo de tamaño mediano en el área de estudio. Se detectó una evidente relación entre el diámetro del disco y la diversidad del tipo de alimento. Los tres grupos de tallas fueron positivamente selectivos hacia corales duros; los más pequeños se alimentaron preferentemente de Porites panamensis y en conjunto con los grandes, presentaron valores bajos de diversidad en el tipo de alimento, mientras que los medianos fueron los que consumieron una mayor diversidad de presas.

Introduction: Little is known about the biology and population dynamics of Acanthaster planci in Mexico. Its size is considered relevant because it is related to consumption rate. Objective: To describe the variability of the disk diameter and food type of the species in four sampling sites in the southern Gulf of California. Methods: Each site was visited on a monthly basis from February 2008 to March 2009. Disk diameter was registered from a total of 389 observed specimens. Measurements included only the main disk from side to side where the arms begin. A flexible tape was used to measure disk diameter. Three size groups were created for easier explanations: small ( 26 cm). From the total, only 298 asteroids were feeding. Preyed organisms were classified in hard corals, other invertebrates, and algae. The Simpson's Diversity Index was used to calculate the probability that any specimen would be feeding on different types of organisms. A permutational analysis of variance was used to detect differences of disk diameter amongst months and study sites. Results: This population of Acanthaster planci had an average disk diameter = 18.23 ± 0.21 cm, with a minimum of 7 cm and maximum of 36 cm. Size structure was unimodal and largely dominated by medium sized specimens, followed by small ones. The larger were less frequent. Disk diameter was statistically higher in February and lower in the last months of the year. San Rafaelito was the site with the largest and Ensenada de Muertos with the smallest organisms. Medium sized individuals were dominant throughout the year and sampling sites. The small sized fed significantly more on Porites panamensis and coralline algae and turf. Larger specimens had the lowest diversity values, more than 90 % of this size group fed on Pocillopora spp. and very few on macroalgae. Medium sized specimens showed the highest diversity, feeding on all the food types, although as expected, many of them were selective to Pocillopora spp. Conclusions: Acanthaster planci can be considered a medium sized species in the study area. There is an evident food selectivity regarding disk diameter size. The three size groups were positively selective to hard corals, but only the small favored Porites panamensis; and together with large individuals, they had the lowest diversity on food types, whereas medium sized asteroids fed on a high diversity of prey.

Population size structure and abnormalities in the number of rays of the Sea Star Pentaceraster cumingi (Valvatida: Oreasteridae) in Bahía Chamela, Mexican Pacific / Estructura de tallas poblacional y anormalidades en el número de radios de la estrella de mar Pentaceraster cumingi (Valvatida: Oreasteridae) en Bahía Chamela, Pacífico mexicano

Introduction: The Panamic Cushion Star Pentaceraster cumingi is widely distributed along the Tropical Eastern Pacific. This species strictly produces only five arms, but sometimes, this number varies or show another kind of abnormality. Objective: We aimed to evaluate the population size structure and abnormalities occurrence in the radial pattern of P. cumingi in Bahía Chamela, Jalisco, Mexico. Methods: The population was monitored along four years (2016-2019), in two seasonal periods (warm and cold). During fieldwork, a random sample of individuals was collected. Every starfish was measured, weighted, and evaluated to identify any abnormality on its radial pattern. Results: The highest density of P. cumingi was found in October 2019 (2.03 ± 0.05 ind/m-2), the lower in March 2017 (0.66 ± 0.13 ind/m-2). A total of 849 individuals were collected. For 5-armed starfishes, the average length was 123.8 ± 15.2 mm and the average weight of 326.0 ± 62.4 g. The most frequent length classes ranged from 110 to 120 mm. Of the total of individuals sampled 0.82 % had four arms, 1.06 % six, and 1.41 % had one bifurcated arm. Conclusions: There were differences in the population density and size structure of P. cumingi between seasons. The main causes of abnormalities in the starfish could be due to the changes that occur during larval metamorphosis or by an abnormal regeneration of the arms after a predation attempt.

Introducción: La Estrella Cojín Pentaceraster cumingi está ampliamente distribuida a lo largo del Pacífico Oriental Tropical. Esta especie produce estrictamente cinco brazos, pero en algunas ocasiones el número puede ser menor o mayor de cinco, o mostrar otro tipo de anormalidad. Objetivo: Evaluar la estructura de tallas poblacional y la presencia de anormalidades en el patrón radial de P. cumingi en la Bahía Chamela, Jalisco, México. Métodos: La población fue monitoreada a lo largo de cuatro años (2016-2019), durante dos periodos estacionales (cálido y templado). Durante el trabajo de campo se recolectó una muestra aleatoria de individuos. Cada estrella de mar fue medida, pesada y revisada para identificar alguna anormalidad en el patrón radial. Resultados: La mayor densidad de individuos se encontró en octubre de 2019 (2.03 ± 0.05 ind/m-2), la menor en marzo de 2017 (0.66 ± 0.13 ind/m-2). Un total de 849 individuos fueron recolectados. Para las estrellas de mar con 5 brazos, la longitud promedio fue de 123.8 ± 15.2 mm y el peso promedio de 326.0 ± 62.4 g. La clase de talla más frecuente varió entre 110 y 120 mm. Del total de individuos muestreados el 0.82 % tuvieron cuatro brazos, el 1.06 % con seis, y el 1.41 % tuvieron un brazo bifurcado. Conclusiones: Se encontraron diferencias en la densidad poblacional y en la estructura de tallas de P. cumingi entre estaciones. Las principales causas de anormalidades en la estrella de mar pueden ser debidas a los cambios que ocurren durante la metamorfosis de las larvas o por una regeneración anormal de los brazos a causa de la depredación.

Diet flexibility and growth of the early herbivorous juvenile crown-of-thorns sea star, implications for its boom-bust population dynamics.

The ecology of the early herbivorous juvenile stage of the crown-of-thorns sea star (COTS, Acanthaster spp.) is poorly understood, yet the success of this life stage is key to generating population outbreaks that devastate coral reefs. Crustose coralline algae (CCA) has been considered to be the main diet of herbivorous juveniles. In this study, we show that COTS can avail of a range of algal food. Juveniles were reared on CCA, Amphiroa sp., and biofilm, and survived for 10 months on all three diets. The juveniles fed CCA and Amphiroa sp. reached 15-16.5 mm diameter at ~ 6 months and maintained this size for the rest the experiment (an additional ~4 months). Juveniles fed biofilm grew more slowly and to a smaller maximum size (~3 mm diameter). However, when juveniles were switched from biofilm to CCA they resumed growth to a new asymptotic size (~13.5 mm, 13-20 months). In diet choice experiments, juveniles did not show a preference between Amphiroa sp. and CCA, but generally avoided biofilm. Our results show that juvenile COTS grew equally well on CCA and Amphiroa sp. and can subsist on biofilm for months. Some juveniles, mostly from the biofilm diet treatment, decreased in size for a time and this was followed by recovery. Flexibility in diet, growth, and prolonged maintenance of asymptotic size indicates capacity for growth plasticity in herbivorous juvenile COTS. There is potential for juvenile COTS to persist for longer than anticipated and increase in number as they wait for the opportunity to avail of coral prey. These findings complicate our ability to predict recruitment to the corallivorous stage and population outbreaks following larval settlement and the ability to understand the age structure of COTS populations.

Relative efficacy of three approaches to mitigate Crown-of-Thorns Starfish outbreaks on Australia's Great Barrier Reef.

Population outbreaks of Crown-of-Thorns Starfish (COTS; Acanthaster spp.) are a major contributor to loss of hard coral throughout the Indo-Pacific. On Australia's Great Barrier Reef (GBR), management interventions have evolved over four COTS outbreaks to include: (1) manual COTS control, (2) Marine Protected Area (MPA) zoning, and, (3) water quality improvement. Here we evaluate the contribution of these three approaches to managing population outbreaks of COTS to minimize coral loss. Strategic manual control at sites reduced COTS numbers, including larger, more fecund and damaging individuals. Sustained reduction in COTS densities and improvements in hard coral cover at a site were achieved through repeated control visits. MPAs influenced initial COTS densities but only marginally influenced final hard coral cover following COTS control. Water quality improvement programs have achieved only marginal reductions in river nutrient loads delivered to the GBR and the study region. This, a subsequent COTS outbreak, and declining coral cover across the region suggest their contributions are negligible. These findings support manual control as the most direct, and only effective, means of reducing COTS densities and improving hard coral cover currently available at a site. We provide recommendations for improving control program effectiveness with application to supporting reef resilience across the Indo-Pacific.

Transcriptomic analysis of sea star development through metamorphosis to the highly derived pentameral body plan with a focus on neural transcription factors.

The Echinodermata is characterized by a secondarily evolved pentameral body plan. While the evolutionary origin of this body plan has been the subject of debate, the molecular mechanisms underlying its development are poorly understood. We assembled a de novo developmental transcriptome from the embryo through metamorphosis in the sea star Parvulastra exigua. We use the asteroid model as it represents the basal-type echinoderm body architecture. Global variation in gene expression distinguished the gastrula profile and showed that metamorphic and juvenile stages were more similar to each other than to the pre-metamorphic stages, pointing to the marked changes that occur during metamorphosis. Differential expression and gene ontology (GO) analyses revealed dynamic changes in gene expression throughout development and the transition to pentamery. Many GO terms enriched during late metamorphosis were related to neurogenesis and signalling. Neural transcription factor genes exhibited clusters with distinct expression patterns. A suite of these genes was up-regulated during metamorphosis (e.g. Pax6, Eya, Hey, NeuroD, FoxD, Mbx, and Otp). In situ hybridization showed expression of neural genes in the CNS and sensory structures. Our results provide a foundation to understand the metamorphic transition in echinoderms and the genes involved in development and evolution of pentamery.

Oocyte Maturation in Starfish.

Oocyte maturation is a process that occurs in the ovaries, where an immature oocyte resumes meiosis to attain competence for normal fertilization after ovulation/spawning. In starfish, the hormone 1-methyladenine binds to an unidentified receptor on the plasma membrane of oocytes, inducing a conformational change in the heterotrimeric GTP-binding protein α-subunit (Gα), so that the α-subunit binds GTP in exchange of GDP on the plasma membrane. The GTP-binding protein ßγ-subunit (Gßγ) is released from Gα, and the released Gßγ activates phosphatidylinositol-3 kinase (PI3K), followed by the target of rapamycin kinase complex2 (TORC2) and 3-phosphoinositide-dependent protein kinase 1 (PDK1)-dependent phosphorylation of serum- and glucocorticoid-regulated kinase (SGK) of ovarian oocytes. Thereafter, SGK activates Na+/H+ exchanger (NHE) to increase the intracellular pH (pHi) from ~6.7 to ~6.9. Moreover, SGK phosphorylates Cdc25 and Myt1, thereby inducing the de-phosphorylation and activation of cyclin B-Cdk1, causing germinal vesicle breakdown (GVBD). Both pHi increase and GVBD are required for spindle assembly at metaphase I, followed by MI arrest at pHi 6.9 until spawning. Due to MI arrest or SGK-dependent pHi control, spawned oocytes can be fertilized normally.

Citizen Science, a promising tool for detecting and monitoring outbreaks of the crown-of-thorns starfish Acanthaster spp.

Monitoring potentially devastating coral-eating crown-of-thorns starfish (COTS) populations at scales relevant to management is a challenging task. Here, we investigated a citizen science approach to detect COTS outbreaks and prioritize management responses. Between 2014 and 2018, 38 000 COTS were recorded through 641 online observation reports submitted across New Caledonia, Vanuatu and Fiji by private stakeholders (51%), NGOs (22%), business operator (11%), research/government agencies (16%). COTS were observed in multiple areas, including in remote/inhabited reefs where they had never been reported. A three-level classification was developed to discriminate risk areas and propose operational guidelines to streamline management actions. About two-thirds of reports had low abundances (<10 starfish sighted) and could be addressed with low priority. Verification surveys at 65 reef sites confirmed outbreaks in half of the cases, along with high peak densities (7 000 ind.ha-1). Combining professional and non-professional observers increased the detection range (+27%) and the number of COTS detections (+129%). Citizen reports were eventually followed by removal campaigns organized within diverse institutional frameworks. While citizen monitoring has intrinsic limitations, we advocate that it constitutes a complementary and promising approach to support the ongoing management efforts in all countries affected by COTS.

Developmental transcriptomes of the sea star, Patiria miniata, illuminate how gene expression changes with evolutionary distance.

Understanding how changes in developmental gene expression alter morphogenesis is a fundamental problem in development and evolution. A promising approach to address this problem is to compare the developmental transcriptomes between related species. The echinoderm phylum consists of several model species that have significantly contributed to the understanding of gene regulation and evolution. Particularly, the regulatory networks of the sea star, Patiria miniata (P. miniata), have been extensively studied, however developmental transcriptomes for this species were lacking. Here we generated developmental transcriptomes of P. miniata and compared these with those of two sea urchins species. We demonstrate that the conservation of gene expression depends on gene function, cell type and evolutionary distance. With increasing evolutionary distance the interspecies correlations in gene expression decreases. The reduction is more severe in the correlations between morphologically equivalent stages (diagonal elements) than in the correlation between morphologically distinct stages (off-diagonal elements). This could reflect a decrease in the morphological constraints compared to other constraints that shape gene expression at large evolutionary divergence. Within this trend, the interspecies correlations of developmental control genes maintain their diagonality at large evolutionary distance, and peak at the onset of gastrulation, supporting the hourglass model of phylotypic stage conservation.

Discovery of Adults Linked to Cloning Oceanic Starfish Larvae (Oreaster, Asteroidea: Echinodermata).

Two juvenile specimens of a new species of Oreaster were collected at Parque Nacional Arrecife Alacranes and Triángulos Oeste in the southern Gulf of Mexico. DNA of mitochondrial loci identifies them as members of the same clade as cloning larvae of Oreaster found abundantly in waters of the Florida Current-Gulf Stream system, and distinct from Oreaster clavatus and Oreaster reticulatus, the two known Oreasteridae species in the North Atlantic. Larvae from the new species of Oreaster persist as clones but also metamorphose and settle to the benthos with typical asteroid morphology.

Abbreviated Development of the Brooding Brittle Star Ophioplocus esmarki.

The bilaterally symmetrical, feeding larval stage is an ancestral condition in echinoderms. However, many echinoderms have evolved abbreviated development and form a pentamerous juvenile without a feeding larva. Abbreviated development with a non-feeding vitellaria larva is found in five families of brittle stars, but very little is known about this type of development. In this study, the external anatomy, ciliary bands, neurons, and muscles were examined in the development of the brooded vitellaria larva of Ophioplocus esmarki. The external morphology throughout development shows typical vitellaria features, including morphogenetic movements to set up the vitellaria body plan, an anterior preoral lobe, a posterior lobe, transverse ciliary bands, and development of juvenile structures on the mid-ventral side. An early population of neurons forms at the base of the preoral lobe at the pre-vitellaria stage after the initial formation of the coelomic cavities. These early neurons may be homologous to the apical neurons that develop in echinoderms with feeding larval forms. Neurons form close to the ciliary bands, but the vitellaria larva lacks the tracts of neurons associated with the ciliary bands found in echinoderms with feeding larvae. Additional neurons form in association with the axial complex and persist into the juvenile stage. Juvenile nerves and muscles form with pentamerous symmetry in the late vitellaria stage in a manner similar to their development within the late ophiopluteus larva. Even though O. esmarki is a brooding brittle star, its developmental sequence retains the general vitellaria shape and structure; however, the vitellaria larvae are unable to swim in the water column.

The impact of ocean acidification on the gonads of three key Antarctic benthic macroinvertebrates.

CO2 atmospheric pressure is increasing since industrial revolution, leading to a lowering of the ocean surface water pH, a phenomenon known as ocean acidification, with several reported effects on individual species and cascading effects on marine ecosystems. Despite the great amount of literature on ocean acidification effects on calcifying organisms, the response of their reproductive system still remains poorly known. In the present study, we investigated the histopathological effects of low pH on the gonads of three key macroinvertebrates of the Terra Nova Bay (Ross Sea) littoral area: the sea urchin Sterechinus neumayeri, the sea star Odontaster validus and the scallop Adamussium colbecki. After 1 month of exposure at control (8.12) and reduced (7.8 and 7.6) pH levels, we dissected the gonads and performed histological analyses to detect potential differences among treatments. Results showed significant effects on reproductive conditions of A. colbecki and S. neumayeri, while O. validus did not show any kind of alteration. Present results reinforce the need to focus on ocean acidification effects on soft tissues, particularly the gonads, whose damage may exert large effects on the individual fitness, with cascading effects on the population dynamic of the species.

Contributions of pre- versus post-settlement processes to fluctuating abundance of crown-of-thorns starfishes (Acanthaster spp.).

Numerous hypotheses have been put forward to account for population outbreaks of crown-of-thorns starfishes (CoTS, Acanthaster spp.), which place specific importance on either pre- or post-settlement mechanisms. The purpose of this review is to specifically assess the contributions of pre- versus post-settlement processes in the population dynamics of CoTS. Given the immense reproductive potential of CoTS (>100 million eggs per female), persistent high densities would appear inevitable unless there were significant constraints on larval development, settlement success, and/or early post-settlement growth and survival. In terms of population constraints, pre- and post-settlement processes are both important and have additive effects to suppress densities of juvenile and adult CoTS within reef ecosystems. It is difficult, however, to assess the relative contributions of pre- versus post-settlement processes to population outbreaks, especially given limited data on settlement rates, as well as early post-settlement growth and mortality. Prioritising this research is important to resolve potential effects of anthropogenic activities (e.g., fishing) and habitat degradation on changing population dynamics of CoTS, and will also improve management effectiveness.

Disassembly of Subplasmalemmal Actin Filaments Induces Cytosolic Ca2+ Increases in Astropecten aranciacus Eggs.

BACKGROUND/AIMS: Eggs of all animal species display intense cytoplasmic Ca2+ increases at fertilization. Previously, we reported that unfertilized eggs of Astropecten aranciacus exposed to an actin drug latrunculin A (LAT-A) exhibit similar Ca2+ waves and cortical flashes after 5-10 min time lag. Here, we have explored the molecular mechanisms underlying this unique phenomenon. METHODS: Starfish eggs were pretreated with various agents such as other actin drugs or inhibitors of phospholipase C (PLC), and the changes of the intracellular Ca2+ levels were monitored by use of Calcium Green in the presence or absence of LAT-A. The concomitant changes of the actin cytoskeleton were visualized with fluorescent F-actin probes in confocal microscopy. RESULTS: We have shown that the LAT-A-induced Ca2+ increases are related to the disassembly of actin flaments: i) not only LAT-A but also other agents depolymerizing F-actin (i.e. cytochalasin B and mycalolide B) induced similar Ca2+ increases, albeit with slightly lower efficiency; ii) drugs stabilizing F-actin (i.e. phalloidin and jasplakinolide) either blocked or significantly delayed the LAT-A-induced Ca2+ increases. Further studies utilizing pharmacological inhibitors of PLC (U-73122 and neomycin), dominant negative mutant of PLC-É£, specific sequestration of PIP2 (RFP-PH), InsP3 uncaging, and quantitation of endogenous InsP3 all indicated that LAT-A induces Ca2+ increases by stimulating PLC rather than sensitizing InsP3 receptors. In support of the idea, it bears emphasis that LAT-A timely increased intracellular contents of InsP3 with concomitant decrease of PIP2 levels in the plasma membrane. CONCLUSION: Taken together, our results suggest that suboolemmal actin filaments may serve as a scaffold for cell signaling and modulate the activity of the key enzyme involved in intracellular Ca2+ signaling.

Effects of larvae density and food concentration on Crown-of-Thorns seastar (Acanthaster cf. solaris) development in an automated flow-through system.

Coral-eating Crown-of-Thorns Sea stars (Acanthaster spp.) are major contributors to coral reef loss in the Indo-Pacific region. A release from food limitation of their planktotrophic larvae through enhanced pelagic productivity is one of the main hypothesis explaining population outbreaks ('nutrient limitation hypothesis'). To improve the understanding of these outbreaks we developed an automated flow- through larvae rearing system that maintained food (microalgae) at set levels over the course of four 15d experiments. This resulted in stable food concentrations in experimental tanks. Increased algae concentrations had a significant positive effect on larval development and size at 10 and 15 days post fertilization (dpf). Larvae densities had no effect at 10 dpf. At 15 dpf greater larvae densities were associated with declines in larvae size. Larval development was slowed under higher larvae densities. Thus, the effects of algae concentration and larvae density were additive at 15 dpf, with larvae under low densities at a given algae concentration being further developed than those under higher densities. The development of a flow-through system gives greater insight into the effect of algae and larvae concentrations on Acanthaster development, and the system can be applied to further test the nutrient-limitation hypothesis for present and future outbreaks.

Chemical Ecology of Chemosensation in Asteroidea: Insights Towards Management Strategies of Pest Species.

Within the Phylum Echinodermata, the class Asteroidea, commonly known as starfish and sea stars, encompasses a large number of benthos inhabiting genera and species with various feeding modalities including herbivores, carnivores, omnivores and detritivores. The Asteroidea rely on chemosensation throughout their life histories including hunting prey, avoiding or deterring predators, in the formation of spawning aggregations, synchronizing gamete release and targeting appropriate locations for larval settlement. The identities of many of the chemical stimuli that mediate these physiological and behavioural processes remain unresolved even though evidence indicates they play pivotal roles in the functionality of benthic communities. Aspects of chemosensation, as well as putative chemically-mediated behaviours and the molecular mechanisms of chemoreception, within the Asteroidea are reviewed here, with particular reference to the coral reef pest the Crown-of-Thorns starfish Acanthaster planci species complex, in the context of mitigation of population outbreaks.

Enzyme-linked immunosorbent assay of relaxin-like gonad-stimulating peptide in the starfish Patiria (Asterina) pectinifera.

A relaxin-like gonad-stimulating peptide (RGP) from starfish Patiria (Asterina) pectinifera is the first identified invertebrate gonadotropin for final gamete maturation. Recently, we succeeded in obtaining specific antibodies against P. pectinifera RGP (PpeRGP). In this study, the antibodies were used for the development of a specific and sensitive enzyme-linked immunosorbent assay (ELISA) for the measurement of PpeRGP. A biotin-conjugated peptide that binds to peroxidase-conjugated streptavidin is specifically detectable using 3,3',5,5'-tetramethylbenzidine (TMB)/hydrogen peroxide as a substrate; therefore, biotin-conjugated RGP (biotin-PpeRGP) was synthesized chemically. Similarly to PpeRGP, synthetic biotin-PpeRGP bound to the antibody against PpeRGP. In binding experiments with biotin-PpeRGP using wells coated with the antibody, a displacement curve was obtained using serial concentrations of PpeRGP. The ELISA system showed that PpeRGP could be measured in the range 0.01-10pmol per 50µl assay buffer. On the contrary, the B-chains of PpeRGP, Asterias amurensis RGP, Aphelasterias japonica RGP, and human relaxin showed minimal cross-reactivity in the ELISA, except that the A-chain of PpeRGP affected it slightly. These results strongly suggest that this ELISA system is highly specific and sensitive with respect to PpeRGP.

Superstars: Assessing nutrient thresholds for enhanced larval success of Acanthaster planci, a review of the evidence.

Crown-of-thorns starfish, Acanthaster planci (COTS), predation is a major cause of coral reef decline, but the factors behind their population outbreaks remain unclear. Increased phytoplankton food resulting from eutrophication is suggested to enhance larval survival. We addressed the hypothesis that larval success is associated with particular chl-a levels in tightly controlled larval:algal conditions. We used chl-a conditions found on coral reefs (0.1-5.0µgchl-aL-1), including nominal threshold levels for disproportionate larval success (≥1.0µgchl-aL-1). High success to the juvenile occurred across an order of magnitude of chl-a concentrations (0.5-5.0µgchl-aL-1), suggesting there may not be a narrow value for optimal success. Oligotrophic conditions (0.1µgchl-aL-1) appeared to be a critical limit. With a review of the evidence, we suggest that opportunistic COTS larvae may be more resilient to low food levels than previously appreciated. Initiation of outbreak populations need not require eutrophic conditions.