Donal T. Manahan

Larval dispersal potential of the tubeworm Riftia pachyptila at deep-sea hydrothermal vents

Hydrothermal vents are ephemeral because of frequent volcanic and tectonic activities associated with crust formation. Although the larvae of hydrothermal vent fauna can rapidly colonize new vent sites separated by tens to hundreds of kilometres, the mechanisms by which these larvae disperse and recruit are not understood. Here we integrate physiological, developmental and hydrodynamic data to estimate the dispersal potential of larvae of the giant tubeworm Riftia pachyptila. At in situ temperatures and pressures (2 °C and 250 atm), we estimate that the metabolic lifespan for a larva of R. pachyptila averages 38 days. In the measured flow regime at a fast-spreading ridge axis (9° 50′ N; East Pacific Rise), this lifespan results in potential along-ridge dispersal distances that rarely exceed 100 km. This limited dispersal results not from the physiological performance of the embryos and larvae, but instead from transport limitations imposed by periodic reversals in along-ridge flows and sustained episodes of across-ridge flow. The lifespan presented for these larvae can now be used to predict dispersal under current regimes at other hydrothermal vent sites.

Energy Metabolism and Amino Acid Transport During Early Development of Antarctic and Temperate Echinoderms

The rates of oxygen consumption by embryos of antarctic echinoderms (Acodontaster hodgsoni, Odontaster validus, Psilaster charcoti, and Sterechinus neumayeri) were compared to the biomas (ash-free dry organic weight) of the egg of each species. These species could survive for months to years (range: 10 months to 5 years) by relying solely on the reserves present in the egg. However, certain species did not use any of the eggs reserves during early development. Embryonic stages of O. validus (a species with planktotrophic larvae) did not decrease in lipid, protein, or total biomass during the first 35 days of development. During the first 42 days of development, embryos of A. hodgsoni (a species with lecithotrophic development) used protein as an energy source. For both species lipid composed 40 to 50% of egg biomass, but was not used as an energy reserve. Larvae of O. validus have a high-affinity transport system for amino acids dissolved in seawater (K1 = 1.3 {mu}M for alanine). The rate of alanine transport from a low concentration (50 nM) could supply 32% of the larvas metabolic needs. This is a 10-fold higher input to metabolism than was determined (3% at 50 nM) for larvae of a temperate asteroid, Asterina miniata. Larvae of antarctic echinoderms live in an environment where the food supply is low for most of the year. Relative to their metabolic rates, antarctic larvae have larger energy stores and planktotrophic larvae have higher nutrient transport capacities when compared to larvae from temperate regions. These physiological differences allow antarctic larvae to survive for long periods without particulate food.

Experimental ocean acidification alters the allocation of metabolic energy

Significance Anthropogenic emission of CO2 is causing global ocean acidification. For many species, biological responses to acidification often show limited impact at the level of the whole animal. Our integrative studies of whole-organism growth and metabolic rates, rates of protein synthesis and ion transport, enzyme activity, and gene expression show that although the organismal-level impact of acidification on developing sea urchins was minimal, dramatic compensation occurred at the cellular level. Increased rates of synthesis and ion transport resulted in 84% of available energy being allocated to those processes under acidification. Defining the limits of differential energy allocation for the maintenance of critical physiological functions in response to compounding stressors will help provide a mechanistic understanding of resilience potential to environmental change. Energy is required to maintain physiological homeostasis in response to environmental change. Although responses to environmental stressors frequently are assumed to involve high metabolic costs, the biochemical bases of actual energy demands are rarely quantified. We studied the impact of a near-future scenario of ocean acidification [800 µatm partial pressure of CO2 (pCO2)] during the development and growth of an important model organism in developmental and environmental biology, the sea urchin Strongylocentrotus purpuratus. Size, metabolic rate, biochemical content, and gene expression were not different in larvae growing under control and seawater acidification treatments. Measurements limited to those levels of biological analysis did not reveal the biochemical mechanisms of response to ocean acidification that occurred at the cellular level. In vivo rates of protein synthesis and ion transport increased ∼50% under acidification. Importantly, the in vivo physiological increases in ion transport were not predicted from total enzyme activity or gene expression. Under acidification, the increased rates of protein synthesis and ion transport that were sustained in growing larvae collectively accounted for the majority of available ATP (84%). In contrast, embryos and prefeeding and unfed larvae in control treatments allocated on average only 40% of ATP to these same two processes. Understanding the biochemical strategies for accommodating increases in metabolic energy demand and their biological limitations can serve as a quantitative basis for assessing sublethal effects of global change. Variation in the ability to allocate ATP differentially among essential functions may be a key basis of resilience to ocean acidification and other compounding environmental stressors.

Transcriptomic analysis of growth heterosis in larval Pacific oysters (Crassostrea gigas)

Compared with understanding of biological shape and form, knowledge is sparse regarding what regulates growth and body size of a species. For example, the genetic and physiological causes of heterosis (hybrid vigor) have remained elusive for nearly a century. Here, we investigate gene-expression patterns underlying growth heterosis in the Pacific oyster (Crassostrea gigas) in two partially inbred (f = 0.375) and two hybrid larval populations produced by a reciprocal cross between the two inbred families. We cloned cDNA and generated 4.5 M sequence tags with massively parallel signature sequencing. The sequences contain 23,274 distinct signatures that are expressed at statistically nonzero levels and show a highly positively skewed distribution with median and modal counts of 9.25 million and 3 transcripts per million, respectively. For nearly half of these signatures, expression level depends on genotype and is predominantly nonadditive (hybrids deviate from the inbred average). Statistical contrasts suggest ≈350 candidate genes for growth heterosis that exhibit concordant nonadditive expression in reciprocal hybrids; this represents only ≈1.5% of the >20,000 transcripts. Patterns of gene expression, which include dominance for low expression and even underdominance of expression, are more complex than predicted from classical dominant or overdominant explanations of heterosis. Preliminary identification of ribosomal proteins among candidate genes supports the suggestion from previous studies that efficiency of protein metabolism plays a role in growth heterosis.

THE UPTAKE AND METABOLISM OF DISSOLVED AMINO ACIDS BY BIVALVE LARVAE

The rates of uptake and metabolism of 14C-labeled glycine and alanine from sea water into larval oysters, Crassostrea gigas (Thunberg) and mussels, Mytilus edulis L. were determined. Kinetic studies showed that both species have a Kt value of 3-4 µM, indicating that bivalve larvae have amino acid transport mechanisms that function efficiently in natural sea water. The Kt values for larvae are similar to those reported for adult bivalves. However, larvae take up dissolved amino acids at approximately ten times the rate reported for adult bivalves on a gram dry weight basis. This difference in uptake capacity presumably reflects the greater absorptive surface area to volume ratio of a larva. Rates of metabolism of absorbed amino acids by larvae were also rapid. Following a 100 min exposure, oyster larvae incorporated 47% of the glycine into protein and 38% was produced as CO2. In comparison to adults, larval bivalves have a more rapid weight-specific uptake and faster rate of utilizing absorbed amino acids....

Feeding by a “nonfeeding” larva: uptake of dissolved amino acids from seawater by lecithotrophic larvae of the gastropod Haliotis rufescens

Larvae of the red abalone (Haliotis rufescens Swainson) are functionally incapable of capturing particulate foods. The aim of this study was to determine whether these larvae could acquire energy from seawater in the form of dissolved organic material. Trochophore and veliger larvae were shown to acquire energy by transporting dissolved organic material from seawater. Both larval stages took up all classes of amino acids tested. The influx of radiolabeled alanine represented the net substrate flux, as determined by direct chemical measurement for both trochophore and veliger larvae. Although veliger larvae have a transport system to take up taurine from seawater, a net efflux was observed for this amino acid. The release of taurine occurred independently of the presence of either taurine or other amino acids in the medium. Transported alanine was used in both anabolic and catabolic pathways. The percent of 14C-alanine in the trichloroacetic acid-insoluble fraction (macromolecules) of veliger larvae ranged from 21 to 56% of the total radioactivity in the larvae. No lipid biosynthesis was detected from 14C-labeled alanine. Veliger larvae catabolized 15 to 19% of the total alanine taken up and released it as 14CO2. The metabolic rate (oxygen consumption) and the rate of amino acid uptake were both determined for the same group of veliger larvae. The percent contribution that the uptake of amino acids, from a total concentration of 1.6 μM, made to the metabolic demand of abalone larvae ranged from 39 to 70%. Thus, these lecithotrophic larvae are not energetically independent of their environment, a result which differs from the current view of energy allocation to “nonfeeding” larvae.

Quantitative and molecular genetic analyses of heterosis in bivalve molluscs

Abstract Associations of allozyme-heterozygosity with growth and its physiological underpinnings have been well documented for bivalve molluscs. The associations are correlational, however, derived almost entirely from studies of wild-caught juveniles or adults. Such studies cannot resolve alternative genetic explanations of heterosis. Four experimental approaches have recently been made to this problem; (1) a correlational study contrasting allozyme and presumably selectively neutral nuclear DNA polymorphisms; (2) detailed studies of allozyme inheritance in families; (3) a study contrasting the performance of meiosis-I and meiosis-II triploids with diploids and (4) a classical quantitative genetic study of the performance of hybrids produced by crosses among inbred lines. The last approach has uncovered remarkable heterosis in growth and its physiological components, both for the larval and juvenile or adult stages, and has implicated epistasis as a significant cause of this heterosis. More importantly, this approach now permits dissection of heterosis into quantitative trait loci (QTL) mapped by the co-segregation of allozyme and nuclear DNA markers with growth phenotypes in the F2 hybrid and backcross generations.

Sources of energy for increased metabolic demand during metamorphosis of the abalone Haliotis rufescens (Mollusca)

Pelagic, lecithotrophic (nonfeeding) larvae of the red abalone (Haliotis rufescens) settle and subsequently metamorphose into benthic juveniles capable of feeding on particulate food. Thus, metamorphosis must be fueled by either endogenous reserves or a nonparticulate food source such as dissolved organic material (DOM) in seawater. The metabolic rates (measured as oxygen consumption) of abalone larvae were found to increase by an average of 3- to 5-fold from the larva to early juvenile stage. The total cost of development from embryo to juvenile measured for three cultures ranged from 41.6 mJ to 55.0 mJ. Meeting this cost would require 1.3 to 1.7 {mu}g of biomass (ash-free dry mass), which is similar to the initial biomass of the spawned oocyte at 1.36 +/- 0.04 {mu}g (mean of four cultures). However, there was no net loss of biomass during development from the oocyte to the juvenile. The uptake of alanine and glucose from seawater by larvae and juveniles could provide one-third of the organic material required to supply metabolism, even if the transporters were only operating at 20% of their maximum capacity throughout development. For larvae undergoing metamorphosis (between 6- and 9-days-old) the proportion of total metabolic demand supplied using aerobically catabolized biomass was only 39%. The higher metabolic rates of metamorphosis are met only in part by consuming stored endogenous reserves. Concomitant with an increase in mass-specific metabolic rate during metamorphosis, the maximal capacity (Jmax) for the transport of dissolved alanine from seawater increased 3-fold, from 61.2 +/- 1.9 (SE) to 182.0 +/- 49 pmol alanine individual-1 h-1. The majority (range: 61% to 100%) of the energy requirements of larval and early juvenile development of H. rufescens could be supplied by input of DOM from the environment. Measurements of transport rates of amino acids and sugars by these animals, and calculations of the energy input from these substrates, indicate that the cumulative transport of DOM from seawater during development to the early juvenile stage could supply an amount of energy equivalent to the initial maternal endowment of energy reserves to the oocyte of this lecithotrophic species.

Energetics of early development for the sea urchinsStrongylocentrotus purpuratus andLytechinus pictus and the crustaceanArtemia sp.

Embryos and larvae of two species of sea urchin,Strongylocentrotus purpuratus andLytechinus pictus, and larvae of the brine shrimpArtemia sp. (San Francisco brand) were cultured to investigate the contribution of dissolved organic material in seawater to the energetics of early development. When embroys ofS. purpuratus were reared in artificial seawater, a net loss in dry organic mass was observed. In contrast, when sibling embryos were reared to Day 2 under identical conditions in natural seawater, there was either a net increase in dry organic mass or no change. A net decrease in mass was observed in only one of five cultures reared in filtered natural seawater. Energy budgets for each species were determined by giving energy equivalents to the changes in carbohydrate, lipid and protein, and to the rate of oxygen consumption for each day of development. In the case ofS. purpuratus, the use of endogenous reserves accounted for either 0 or 38% of the metabolic demand for two independent cultures reared from Days 0 to 2. For larvae ofL. pictus, reared to 8 d, only 66% of the metabolic demand could be accounted for by the use of endogenous reserves. Sea urchins are capable of transporting dissolved organic material from seawater. Calculations revealed that the energy deficit during the early development of sea urchins (S. purpuratus) could be accounted for by the uptake of dissolved organic matter from seawater. However, for a species that cannot use this resource (Artemia sp.), the metabolic needs during development are supplied through the use of endogenous reserves.

Energy metabolism during larval development of green and white abalone, Haliotis fulgens and H. sorenseni

An understanding of the biochemical and physiological energetics of lecithotrophic development is useful for interpreting patterns of larval development, dispersal potential, and life-history evolution. This study investigated the metabolic rates and use of biochemical reserves in two species of abalone, Haliotis fulgens (the green abalone) and H. sorenseni (the white abalone). Larvae of H. fulgens utilized triacylglycerol as a primary source of endogenous energy reserves for development (∼50% depletion from egg to metamorphic competence). Amounts of phospholipid remained constant, and protein dropped by about 30%. After embryogenesis, larvae of H. fulgens had oxygen consumption rates of 81.7 ± 5.9 (SE) pmol larva−1 h−1 at 15 °C through subsequent development. The loss of biochemical reserves fully met the needs of metabolism, as measured by oxygen consumption. Larvae of H. sorenseni were examined during later larval development and were metabolically and biochemically similar to H. fulgens larvae at a comparable stage. Metabolic rates of both species were very similar to previous data for a congener, H. rufescens, suggesting that larval metabolism and energy utilization may be conserved among closely related species that also share similar developmental morphology and feeding modes.