Understanding the Symbiotic Relationship between the Sea Urchin Loxechinus albus (Molina, 1782) and the Pea Crab Pinnaxodes chilensis (H. Milne Edwards, 1837): a Potential Parasitism.

The echinoderm Loxechinus albus has a symbiotic relationship with the pinnotherid crustacean Pinnaxodes chilensis. Females of the crustacean develop in the terminal section of the sea urchin's digestive system, remaining there for life. This relationship has been suggested as commensalism. However, a potential negative impact on gonadal development and on the morphology of the sea urchin's digestive system suggest that it is instead parasitic. To study if there is a negative impact of the crustacean symbiont on the host, specimens of L. albus of all sizes were collected from a rocky shore in southern Chile. The gonadal and somatic tissues of sea urchins that were and were not harboring the pinnotherid were weighed and compared. Our results show that the presence of the pinnotherid was related to sea urchin gonads of lower biomass, decreased gonadosomatic index levels, and alterations in the morphology of the terminal portion of the host digestive system. The lower gonadal biomass suggests a negative impact on gamete production as well as a diversion of energy due to changes of the digestive system tissues and the potential consumption of algal food by the resident crustacean. These results suggest that the prolonged relationship between these two species is one of parasitism rather than one of commensalism.

Body mass index does not decline during winter for the sedentary marine gastropod Crepidula fornicata.

Seasonal extremes in environmental conditions can substantially limit the growth and reproduction of animals. Sedentary marine animals are particularly susceptible to winter food limitation since they cannot relocate to more favourable conditions. In several temperate-zone bivalve species, substantial winter tissue mass declines have been documented; however, no comparable studies have been conducted on intertidal gastropods. Here, we investigate whether the suspension-feeding intertidal gastropod Crepidula fornicata also loses substantial tissue mass during the winter. We calculated body mass index (BMI) for individuals collected in New England at different times of year for 7 years to determine whether BMI declines through winter or varies seasonally. Remarkably, C. fornicata body mass did not decline significantly during winter months; indeed, a relatively poorer body condition was associated with higher seawater temperature, higher air temperature and higher chlorophyll concentration. In a laboratory experiment, we found that C. fornicata adults that were not fed for three weeks at 6°C (local winter seawater temperature) showed no detectable declines in BMI compared to field-collected individuals. Future studies should document energy budgets of C. fornicata and other sedentary marine animals at low winter seawater temperatures, and the impact of short-term elevated temperatures on those energy budgets.

The Marine Gastropod Crepidula fornicata Remains Resilient to Ocean Acidification Across Two Life History Stages.

Rising atmospheric CO2 reduces seawater pH causing ocean acidification (OA). Understanding how resilient marine organisms respond to OA may help predict how community dynamics will shift as CO2 continues rising. The common slipper shell snail Crepidula fornicata is a marine gastropod native to eastern North America that has been a successful invader along the western European coastline and elsewhere. It has also been previously shown to be resilient to global change stressors. To examine the mechanisms underlying C. fornicata's resilience to OA, we conducted two controlled laboratory experiments. First, we examined several phenotypes and genome-wide gene expression of C. fornicata in response to pH treatments (7.5, 7.6, and 8.0) throughout the larval stage and then tested how conditions experienced as larvae influenced juvenile stages (i.e., carry-over effects). Second, we examined genome-wide gene expression patterns of C. fornicata larvae in response to acute (4, 10, 24, and 48 h) pH treatment (7.5 and 8.0). Both C. fornicata larvae and juveniles exhibited resilience to OA and their gene expression responses highlight the role of transcriptome plasticity in this resilience. Larvae did not exhibit reduced growth under OA until they were at least 8 days old. These phenotypic effects were preceded by broad transcriptomic changes, which likely served as an acclimation mechanism for combating reduced pH conditions frequently experienced in littoral zones. Larvae reared in reduced pH conditions also took longer to become competent to metamorphose. In addition, while juvenile sizes at metamorphosis reflected larval rearing pH conditions, no carry-over effects on juvenile growth rates were observed. Transcriptomic analyses suggest increased metabolism under OA, which may indicate compensation in reduced pH environments. Transcriptomic analyses through time suggest that these energetic burdens experienced under OA eventually dissipate, allowing C. fornicata to reduce metabolic demands and acclimate to reduced pH. Carry-over effects from larval OA conditions were observed in juveniles; however, these effects were larger for more severe OA conditions and larvae reared in those conditions also demonstrated less transcriptome elasticity. This study highlights the importance of assessing the effects of OA across life history stages and demonstrates how transcriptomic plasticity may allow highly resilient organisms, like C. fornicata, to acclimate to reduced pH environments.

Impact of short-term elevated temperature stress on winter-acclimated individuals of the marine gastropod Crepidula fornicata.

The intertidal zone is an especially stressful thermal habitat, typically exposing residents to air temperatures for up to 6 h at a time, twice daily. Tolerance to elevated temperatures has been particularly well-studied for a variety of intertidal species, especially with regard to upper thermal limits during summers. However, in recent years, as climates have been changing around the world, temperate zone intertidal organisms have sometimes been exposed to periods of unusually high air temperatures during the winter. The present study sought to examine the impact of elevated temperatures on survival and clearance rates of winter-acclimated intertidal individuals of the sedentary marine suspension-feeding gastropod Crepidula fornicata. Individuals were collected intertidally from Nahant, Massachusetts from late January to early April each year for 5 years, maintained in the laboratory at the acclimation temperature of 6 °C, and exposed in the laboratory for 3 h to temperatures as high as 37 °C in seawater either once or twice, 24 h apart. Although mean clearance rates were substantially reduced for at least the next 12-24 h after individuals were returned to the 6 °C control condition following exposures to elevated temperatures as low as 21-26 °C, we saw little mortality even following two 3 h exposures to 35 °C, or single exposures to 37 °C. Mortality was substantial, however, in one experiment following a double exposure to 37 °C. Smaller individuals (~5-12 mm in shell length) were somewhat more sensitive to the thermal stress than adults were. Intertidal members of C. fornicata in Massachusetts seem well-prepared to deal with the increasing range of winter air temperatures associated with the global climate confusion predicted for future years. Additional studies will be required to understand the physiological and biochemical mechanisms used by winter-acclimated individuals of this species to tolerate such periodic substantial temperature increases of 29-31 °C.

Instant Ocean Versus Natural Seawater: Impacts on Aspects of Reproduction and Development in Three Marine Invertebrates.

Marine invertebrate larvae have often been reared in artificial rather than natural seawater, either for convenience or to avoid potentially confounding effects of unknown contaminants. This study sought to determine the impact of artificial seawater on various aspects of development for three marine invertebrate species. We examined the impact of Instant Ocean on growth, survival, and fecundity of the deposit-feeding polychaete Capitella teleta at 2 salinities: 24 and 34 ppt; the impact on survival, growth rate, and time to metamorphic competence for the slipper limpet Crepidula fornicata; and the impact on larval growth for the sea star Asterias forbesi. Juveniles of C. teleta survived better in natural seawater than in Instant Ocean at both salinities but at the higher salinity grew more quickly in Instant Ocean; fecundity was not significantly affected by the type of seawater used at either salinity. Using Instant Ocean in place of natural seawater had no pronounced impact on the survival of C. fornicata larvae or on how long it took them to become competent to metamorphose; however, larvae grew somewhat more quickly in Instant Ocean than in natural seawater for the first 4 days of development, but by day 7 they were about 4.5% larger if they had been reared in seawater. The type of seawater used affected the growth of A. forbesi larvae, with larvae growing significantly more slowly in Instant Ocean than in natural seawater, no matter how growth was measured. In conclusion, our results suggest that although Instant Ocean may be a reasonable substitute for natural seawater for work with some species, using it may affect experimental outcomes in some aspects of work with other species.

Temperature and Salinity Effects on Shell Selection by the Hermit Crab Pagurus longicarpus.

The widespread coastal hermit crab Pagurus longicarpus relies on empty gastropod shells for shelter. At low tide, these hermit crabs often become stranded in tide pools, where changes in temperature and salinity can occur rapidly. We tested how changes in temperature and salinity affected the sizes of the shells chosen by hermit crabs. Increasing the seawater temperature from 22 °C to 32 °C had a significant effect ( P<0.05 ) on the size of shells chosen, as did lowering the temperature to 16 °C. Reducing the salinity from 30 ppt to 20 ppt or raising it to 40 ppt did not affect shell choice significantly, but lowering the salinity from 30 ppt to 15 ppt did. This study builds on previous studies of shell size preference of hermit crabs by highlighting the importance of taking temperature and salinity into account when assigning shells of appropriate size in laboratory studies. These results also suggest that climate change may alter the sizes of shells that hermit crabs of the species P. longicarpus will seek in the field. Additional studies will be required to evaluate the potential impacts of such shifts on hermit crab survival and reproduction.

Growing Safe: Acute Size Escape from Desiccation in Juvenile Crepipatella peruviana (Mollusca: Gastropoda).

Desiccation is an important limiting factor in the intertidal zone. Generally decreasing seaward, desiccation stress can also be alleviated in wet microhabitats. Juvenile snails are generally more susceptible to desiccation than adults, and, for some species, juveniles must therefore hide in microhabitats to survive emersion. The transition from hiding in safe microhabitats to being able to survive fully exposed for the duration of low tide is not well documented. In this study, we investigated the influence of size on desiccation tolerance in juveniles of the calyptraeid gastropod Crepipatella peruviana to determine the size at which they can first survive exposure to air. Juveniles 2-13 mm long were exposed to 75% or 100% relative humidity for 0.5-6.5 hours. Juveniles smaller than 5 mm in shell length did not survive emersion at 75% relative humidity for even 0.5 hours; surprisingly, most also perished after short exposures to air at 100% relative humidity, suggesting that something other than desiccation stress may also be at play. In marked contrast, 82% of juveniles larger than 6 mm in shell length survived exposure to 75% relative humidity for the full 6.5 hours. In a field survey, no juveniles smaller than 9 mm were found on exposed rock but rather were found only in wet microhabitats. We suggest that the clearly defined size escape from desiccation may reflect a change in gill functioning or a newfound ability to retain water more effectively within the mantle cavity at low tide.

Effects of Embryonic Exposure to Salinity Stress or Hypoxia on Post-metamorphic Growth and Survival of the Polychaete Capitella teleta.

Although a good number of studies have investigated the impact of larval experience on aspects of post-metamorphic performance, only a few have considered the potential impact of stresses experienced by brooded embryos. In this study we separately investigated the impact of salinity stress (as low as 10) and hypoxia (1 ml O2 l-1) experienced by brooded embryos of the deposit-feeding polychaete Capitella teleta on hatching success, metamorphosis, post-metamorphic survival, and post-metamorphic growth. Salinity reduction from 30 to 10 or 15 reduced relative hatching success, presumably by reducing embryonic survival, but generally had no negative latent effects on juvenile survival or growth. Prolonged exposure to hypoxic conditions had no negative effects, as seen on measurements recorded, other than abandonment of brood tubes by some females. There were no negative effects on days to emergence from brood tubes, numbers of larvae emerging from brood tubes, juvenile survival, or juvenile growth. Future studies should consider the potential role of maternal behavior in protecting embryos from at least short-term exposures to hypoxia, and the capacity for anaerobic metabolism in both embryos and adults of this species.

Role of the Substrate in Feeding and Growth of the Marine Suspension-Feeding Gastropods Crepidula fornicata and Crepipatella peruviana.

Unlike lamellibranch bivalves, suspension-feeding calyptraeid gastropods lack siphons and paired shell valves to regulate water inflow. This study was designed to determine if calyptraeid gastropods use the solid surface to which they attach to facilitate food particle capture. Juveniles of both Crepidula fornicata and Crepipatella peruviana were maintained with phytoplankton for 3 to 6 wk in the laboratory, either attached to solid substrate or without solid substrate. Individuals of C. fornicata and C. peruviana that were reared on solid substrate grew about five to ten times more, or two times more, respectively, than those deprived of solid substrate. Final tissue weights were also significantly greater for individuals of both species that had been reared on solid substrate. For the two species, phytoplankton clearance rates were about two to three times higher for individuals attached to solid substrate than for those without solid substrate; rates of food cord production from the gills were also significantly higher. About 50% of C. peruviana that were deprived of solid substrate died during the first 3 wk of observation, and about 60% were dead by 6 wk. In contrast, most individuals of C. peruviana that were attached to solid substrate survived for the entire 6-wk study period, and all of the C. fornicata survived whether or not they were attached to solid substrate. The solid substrate to which calyptraeid gastropods attach clearly plays an important role in their feeding biology, although the precise role remains to be explored.

Brooding strategy in fluctuating salinity environments: oxygen availability in the pallial cavity and metabolic stress in females and offspring in the Chilean oyster Ostrea chilensis.

Organisms that encounter stressful situations in nature often cope using behavioral (e.g., avoidance) or physiological tactics. In sessile mollusks, the only available behavioral option in dealing with salinity stress is to "clam up", isolating their tissues from the environment. Though effective in the short term, prolonged isolation can have detrimental physiological consequences, particularly for females brooding embryos in a mantle cavity that is isolated from the external environment. In the Quempillén estuary, the Chilean oyster, Ostrea chilensis, spent nearly one-third of its brooding season at salinities low enough to cause female isolation. When females thus isolated themselves, the dissolved oxygen in their mantle cavity fluid dropped to hypoxic levels within 10 min. In females that were brooding embryos, this depletion of oxygen was not uniform: oxygen was depleted more quickly in the palp region (where embryos accumulate) than in the inhalant region. Additionally, oxygen was reduced even more quickly in the palp region when females were brooding late-stage embryos, which consumed oxygen significantly more quickly than embryos in earlier developmental stages. Finally, O. chilensis used anaerobic metabolism to cope with the hypoxia induced by isolation, as lactate accumulated in the tissues of both females (brooding > non-brooding) and embryos (late stage > early stage). Our findings demonstrate the trade-off between an adaptive avoidance behavior (clamming up) and the potentially detrimental consequences brought on by such a behavior (hypoxia). Cycling of embryos throughout the mantle cavity by deliberate female pumping keeps them from accumulating in the area between the palps, forestalling the creation of hypoxic conditions there. In addition, the capacity for anaerobic metabolism by both females and their embryos should help them tolerate the low oxygen levels that do eventually arise when the pallial cavity is isolated from the surrounding environment during long periods of reduced ambient salinity.

Brooding in the Chilean oyster Ostrea chilensis: unexpected complexity in the movements of brooded offspring within the mantle cavity.

Brooding in invertebrates serves to protect embryos from stressful external conditions by retaining progeny inside the female body, effectively reducing the risk of pelagic stages being exposed to predation or other environmental stressors, but with accompanying changes in pallial fluid characteristics, including reduced oxygen availability. Brooded embryos are usually immobile and often encapsulated, but in some Ostrea species the embryos move freely inside the female pallial cavity in close association with the mother's gills for as long as eight weeks. We used endoscopic techniques to characterize the circulation pattern of embryos brooded by females of the oyster, Ostrea chilensis. Progeny at embryonic and veliger stages typically circulated in established patterns that included the use of dorsal and ventral food grooves (DFG, VFG) to move anteriorly on the gills. Both embryos and veligers accumulated around the mother's palps, and remained there until an active maternal countercurrent moved them to the gill inhalant area. Both food grooves were able to move embryos, veligers, and food-particle aggregates anteriorly, but the DFG was more important in progeny transport; early embryos were moved more rapidly than veligers in the DFG. A microcirculation pattern of embryos was apparent when they were moved by gill lamellae: when they were close to the VFG, most embryos lost gill contact and "fell" down to the DFG. Those that actually reached the DFG moved anteriorly, but others came into contact with the base of the lamellae and again moved towards the VFG. The circulation pattern of the progeny appears well-suited for both cleaning them and directing them posteriorly to an area where there is more oxygen and food than in the palp region. This process for actively circulating progeny involves the feeding structures (gill and palps) and appears to be energetically costly for the female. It also interferes with feeding, which could explain the poor energy balance previously documented for brooding females of this species.

The B vitamins nicotinamide (B3) and riboflavin (B2) stimulate metamorphosis in larvae of the deposit-feeding polychaete Capitella teleta: implications for a sensory ligand-gated ion channel.

Marine sediments can contain B vitamins, presumably incorporated from settled, decaying phytoplankton and microorganisms associated with decomposition. Because B vitamins may be advantageous for the energetically intensive processes of metamorphosis, post-metamorphic growth, and reproduction, we tested several B vitamins to determine if they would stimulate larvae of the deposit-feeding polychaete Capitella teleta to settle and metamorphose. Nicotinamide and riboflavin individually stimulated larvae of C. teleta to settle and metamorphose, generally within 1-2 hours at nicotinamide concentrations as low as 3 µM and riboflavin concentrations as low as 50 µM. More than 80% of the larvae metamorphosed within 30 minutes at a nicotinamide concentration of 7 µM. The pyridine channel agonist pyrazinecarboxamide also stimulated metamorphosis at very low concentrations. In contrast, neither lumichrome, thiamine HCl, pyridoxine HCl, nor vitamin B12 stimulated larvae of C. teleta to metamorphose at concentrations as high as 500 µM. Larvae also did not metamorphose in response to either nicotinamide or pyrazinecarboxamide in calcium-free seawater or with the addition of 4-acetylpyridine, a competitive inhibitor of the pyridine receptor. Together, these results suggest that larvae of C. teleta are responding to nicotinamide and riboflavin via a chemosensory pyridine receptor similar to that previously reported to be present on crayfish chela and involved with food recognition. Our data are the first to implicate B vitamins as possible natural chemical settlement cues for marine invertebrate larvae.

Effects of low salinity on adult behavior and larval performance in the intertidal gastropod Crepipatella peruviana (Calyptraeidae).

Shallow-water coastal areas suffer frequent reductions in salinity due to heavy rains, potentially stressing the organisms found there, particularly the early stages of development (including pelagic larvae). Individual adults and newly hatched larvae of the gastropod Crepipatella peruviana were exposed to different levels of salinity stress (32(control), 25, 20 or 15), to quantify the immediate effects of exposure to low salinities on adult and larval behavior and on the physiological performance of the larvae. For adults we recorded the threshold salinity that initiates brood chamber isolation. For larvae, we measured the impact of reduced salinity on velar surface area, velum activity, swimming velocity, clearance rate (CR), oxygen consumption (OCR), and mortality (LC50); we also documented the impact of salinity discontinuities on the vertical distribution of veliger larvae in the water column. The results indicate that adults will completely isolate themselves from the external environment by clamping firmly against the substrate at salinities ≤24. Moreover, the newly hatched larvae showed increased mortality at lower salinities, while survivors showed decreased velum activity, decreased exposed velum surface area, and decreased mean swimming velocity. The clearance rates and oxygen consumption rates of stressed larvae were significantly lower than those of control individuals. Finally, salinity discontinuities affected the vertical distribution of larvae in the water column. Although adults can protect their embryos from low salinity stress until hatching, salinities <24 clearly affect survival, physiology and behavior in early larval life, which will substantially affect the fitness of the species under declining ambient salinities.

Paternity and gregariousness in the sex-changing sessile marine gastropod Crepidula convexa: comparison with other protandrous Crepidula species.

In sex-changing animals with internal fertilization, gregarious behavior may increase mating opportunities and the frequency of multiple paternity, thus increasing maternal reproductive success. Crepidula convexa is a direct-developing protandrous gastropod characterized by only modest gregarious behavior compared with previously studied members of the genus: females are frequently found isolated. Using 6 microsatellite markers, we analyzed paternity profiles in 10 broods (25 embryos per mother). The number of assigned fathers varied among families from 1 to 4 fathers per brood. Interestingly, polyandry was not detected in solitary females but only in females grouped with conspecific individuals. Overall, we found an average of 1.8 fathers per brood, but this increased to 2.6 fathers per brood when considering only the nonisolated females. Among 18 unambiguously identified fathers, only 5 were collected in our samples, suggesting substantial male mobility. Comparison with previous paternity analyses in Crepidula fornicata and Crepidula coquimbensis revealed that polyandry appears as a common trait of these sex-changing gastropods despite their different grouping behaviors and life histories. As expected, the level of polyandry was nevertheless lower in the modestly gregarious C. convexa.

Understanding the effects of low salinity on fertilization success and early development in the sand dollar Echinarachnius parma.

Free-spawning marine invertebrates that live near shore or in estuaries may experience reduced fertilization success during low-salinity events. Although several studies have documented reproductive failure at reduced salinity in estuarine animals, few have looked at whether developmental failure is due to a failure of fertilization or to a failure of fertilized eggs to cleave. In this study, we examined the effects of salinities ranging from 18 to 32 psu on fertilization success and early development in the sand dollar Echinarachnius parma. In addition to decoupling the effects of low salinity on fertilization from its effects on early cleavage, we also assessed whether eggs or sperm were the weak link in accounting for reproductive failure. We found that both fertilization and cleavage failed at salinities below about 22 psu but that development could be partially rescued by returning zygotes to full-strength seawater. We also found that sperm remained active and capable of fertilizing eggs even after being exposed to low salinities for 30 min.. Taken together, these results suggest that reproductive failure at low salinities in E. parma is due more to an inability of the fertilized eggs to cleave than to an inability of sperm to fertilize eggs.

Fast versus slow larval growth in an invasive marine mollusc: does paternity matter?

Reproductive strategies and parental effects play a major role in shaping early life-history traits. Although polyandry is a common reproductive strategy, its role is still poorly documented in relation to paternal effects. Here, we used as a case study the invasive sessile marine gastropod Crepidula fornicata, a mollusc with polyandry and extreme larval growth variation among sibling larvae. Based on paternity analyses, the relationships between paternal identity and the variations in a major early life-history trait in marine organisms, that is, larval growth, were investigated. Using microsatellite markers, paternities of 437 fast- and slow-growing larvae from 6 broods were reliably assigned to a set of 20 fathers. No particular fathers were found responsible for the specific growth performances of their offspring. However, the range of larval growth rates within a brood was significantly correlated to 1) an index of sire diversity and 2) the degree of larvae relatedness within broods. Multiple paternity could thus play an important role in determining the extent of pelagic larval duration and consequently the range of dispersal distances achieved during larval life. This study also highlighted the usefulness of using indices based on fathers' relative contribution to the progeny in paternity studies.

Nitric oxide inhibits metamorphosis in larvae of Crepidula fornicata, the slippershell snail.

This paper concerns the role of nitric oxide (NO) in controlling metamorphosis in the marine gastropod Crepidula fornicata. Metamorphosis was stimulated by the nitric oxide synthase (NOS) inhibitors AGH (aminoguanidine hemisulfate) and SMIS (S-methylisothiourea sulfate) at concentrations of about 100-1000 micromol l(-1) and 50-200 micromol l(-1), respectively. Metamorphosis was not, however, induced by the NOS inhibitor l-NAME (l-N(G)-nitroarginine methyl ester) at even the highest concentration tested, 500 micromol l(-1). Moreover, pre-incubation with l-NAME at 20 and 80 micromol l(-1) did not increase the sensitivity of competent larvae to excess K(+), a potent inducer of metamorphosis in this species; we suggest that either l-NAME is ineffective in suppressing NO production in larvae of C. fornicata, or that it works only on the constitutive isoform of the enzyme. In contrast, metamorphosis was potentiated by the guanylate cyclase inhibitor ODQ (1H-[1,2,4]oxadiazolo[4,3, -a]quinoxalin-1-one) in response to a natural metamorphic inducer derived from conspecific adults. Because NO typically stimulates cGMP production through the activation of soluble guanylate cyclase, this result supports the hypothesis that NO acts as an endogenous inhibitor of metamorphosis in C. fornicata. The expression of NOS, shown by immunohistochemical techniques, was detected in the apical ganglion of young larvae but not in older larvae, further supporting the hypothesis that metamorphosis in C. fornicata is made possible by declines in the endogenous concentration of NO during development.

Effects of salinity on spawning and early development of the tube-building polychaete Hydroides elegans in Hong Kong: not just the sperm's fault?

Ambient salinities drop dramatically during monsoon season in Hong Kong coastal waters, posing a number of problems for externally fertilizing species like the polychaete Hydroides elegans. In this study, we investigated (1) whether adults would retain their gametes when external salinity dropped to levels too low to support fertilization and development, and (2) whether failure of development at low salinity reflects a failure of fertilization or a failure of fertilized eggs to cleave. Adults released eggs and sperm in the laboratory even at the lowest salinity tested, a practical salinity (S) of 5, and yet very few eggs cleaved at salinities below about 22. By mixing gametes at high salinity and then transferring the fertilized eggs to low-salinity seawater, we found that salinities below about 22 reduced the percentage of fertilized eggs that cleaved. Similarly, mixing gametes at salinities as low as 15 and then transferring the eggs to full-strength seawater (S = 30) rescued a substantial number of eggs, many more of which cleaved after their transfer to the higher salinity. The results suggest that failure of early development at low salinity in this species in large part reflects an inability of newly fertilized eggs to complete meiosis and cleave, rather than simply a failure of fertilization.

Larval experience and latent effects--metamorphosis is not a new beginning.

For many years ecologists have documented the remarkable within-species variation inherent in natural systems-for example, variability in juvenile growth rates, mortality rates, fecundities, time to reproductive maturity, the outcomes of competitive interactions, and tolerance to pollutants. Over the past 20 years, it has become increasingly apparent that at least some of this variation may reflect differences in embryonic or larval experiences. Such experiences may include delayed metamorphosis, short term starvation, short term salinity stress, or exposure to sublethal concentrations of pollutants or sublethal levels of ultra violet irradiation. Latent effects-effects that have their origins in early development but that are first exhibited in juveniles or adults-have now been documented among gastropods, bivalves, echinoderms, polychaetes, crustaceans, bryozoans, urochordates, and vertebrates. The extent to which latent effects alter ecological outcomes in natural populations in the field, and the mechanisms through which they are mediated are largely unexplored.

Timing is everything: the effects of putative dopamine antagonists on metamorphosis vary with larval age and experimental duration in the prosobranch gastropod Crepidula fornicata.

The signal transduction pathway through which excess potassium ion stimulates the larvae of many marine invertebrates to metamorphose is incompletely understood. Recent evidence suggests that dopamine plays important roles in the metamorphic pathway of Crepidula fornicata. Therefore, we asked whether blocking dopamine receptors might prevent excess potassium ion from stimulating metamorphosis in this species. Surprisingly, the effects of the three putative dopamine antagonists tested (all at 10 microM) varied with exposure duration and the age of competent larvae. Chlorpromazine, a nonspecific dopamine antagonist known to have a number of other pharmacological effects, blocked the inductive action of excess potassium ion during the initial 5-8-h exposure periods in most assays, particularly for younger or smaller competent larvae. However, chlorpromazine in the absence of excess potassium ion also stimulated metamorphosis, particularly over the next 18 h, and worked faster on older competent larvae than on younger competent larvae. The specific D(1) antagonist R(+)-Sch-23309 had similar effects, blocking potassium-stimulated metamorphosis in short-term exposures and stimulating metamorphosis in longer exposures, particularly for older competent larvae. Although the specific D(2) antagonist spiperone (SPIP) blocked the inductive effects of excess potassium ion in only 1 of 6 assays during the first 6 h of exposure, it blocked metamorphosis in 2 of the assays during 24-h exposures. Our results indicate that dopamine receptors are involved in the pathway through which excess potassium ion stimulates metamorphosis in C. fornicata. In addition, the largely latent inductive effects of chlorpromazine, an inhibitor of nitric oxide synthase, suggest that endogenous nitric oxide may play a natural role in inhibiting metamorphosis in this species. Overall, our results would then suggest that exposing larvae of C. fornicata to excess K(+) leads to a shutdown of nitric oxide synthesis via a dopaminergic pathway, a pathway that can be blocked by some dopamine antagonists. Alternatively, chlorpromazine might eventually be stimulating metamorphosis by elevating endogenous cyclic nucleotide (e.g., cAMP) concentrations, again acting downstream from the steps acted on directly by excess K(+).