Phenotypic variation in metamorphosis in four species of freshwater copepods

1. Physiological metamorphosis accompanied by an ecological habitat shift is a widespread life-history phenomenon, and both age and size at metamorphosis are highly variable in many organisms. In this study, age and size at metamorphosis (defined as the transition from the last naupliar to the first copepodite stage) were quantified for four species of freshwater copepods to determine the scale on which these two traits vary, if age and size at metamorphosis are equally variable, and if variation at metamorphosis is related to variation in newborn size. 2. Measurements of laboratory-reared and field-caught individuals show that age and size at metamorphosis, together with newborn size, vary among siblings, between families within a population, between populations of one species and between closely related species. 3. In all populations, age at metamorphosis was the most variable trait, a result observed in many other organisms. Most of the variation in age at metamorphosis could be explained by differences between families within a population, while differences among siblings from the same clutch accounted for most of the variation in size at metamorphosis. 4. Although newborn size was variable, differences in this trait could not fully account for variation observed at metamorphosis. Newborn size differed among populations, but most interpopularional differences disappeared by the rime metamorphosis was reached. In particular, size at metamorphosis appears to be tightly constrained in freshwater copepods. 5. Age and size at metamorphosis were not equally variable among species, either. Species-specific metamorphic envelopes (joint distributions of age and size at metamorphosis) result from differences in trait means, variances and covariances, and suggest very different larval growth trajectories among three of the species examined.     

Metamorphosis of cranial design in tiger salamanders (Ambystoma tigrinum): A morphometric analysis of ontogenetic change

While ontogenetic analyses of skull development have contributed to our understanding of phylogenetic patterns in vertebrates, there are few studies of taxa that undergo a relatively discrete and rapid change in morphology during development (metamorphosis). Morphological changes occurring in the head at metamorphosis in tiger salamanders (Ambystoma tigrinum) were quantified by a morphometric analysis of cranial osteology and myology to document patterns of change during metamorphosis. We employed a cross-sectional analysis using a sample of larvae just prior to metamorphosis and a sample of transformed individuals just after metamorphosis, as well as larvae undergoing metamorphosis. There were no differences in external size of the head among the larval and transformed samples. The hyobranchial apparatus showed many dramatic changes at metamorphosis, including shortening of ceratobranchial 1 and the basibranchial. The subarcualis rectus muscle increased greatly in length at metamorphosis, as did hypobranchial length and internasal distance. A truss analysis of dorsal skull shape showed that at metamorphosis the snout becomes wider, the maxillary and squamosal triangles rotate posteromedially, and the neurocranium shortens (while maintaining its width), resulting in an overall decrease in skull length at metamorphosis. These morphometric differences are interpreted in light of recent data on the functional morphology of feeding in salamanders. Morphological reorganization of the hyobranchial apparatus and shape changes in the skull are related to the acquisition of a novel terrestrial feeding mode (tongue projection) at metamorphosis. Metamorphic changes (both internal and external) that can be used to judge metamorphic condition are discussed.     

黑斑蛙变态过程中甲状腺发育及甲状腺激素分泌

系统研究了我国本土两栖动物种黑斑蛙（Rana nigromaculata）变态发育过程中甲状腺组织学和甲状腺激素水平的变化,为甲状腺生物学和甲状腺干扰研究提供基础数据。黑斑蛙蝌蚪发育的形态变化: 第26-40阶段,后腿芽生长并逐渐分化出五趾结构;42阶段,开始进入变态高峰期,前肢展开,尾吸收,蝌蚪身体发生巨大形变;46阶段,蝌蚪完全变态成小蛙。随着形态学的变化,甲状腺的组织结构也发生明显的变化: 26-37阶段,甲状腺体积较小,增长缓慢;38阶段甲状腺体积迅速膨大,进入高峰期,甲状腺的发育达到顶峰;随着变态完成,甲状腺又逐渐缩小。甲状腺组织学变化的同时,甲状腺激素水平也相应发生变化: 在变态前期,下颌中3,3',5-三碘代-L-甲腺原氨酸（T3）水平增长缓慢,进入变态期后,T3含量迅速升高,在变态高峰期达到峰值,随后下降。以上结果表明,黑斑蛙发育过程中甲状腺组织学的变化与甲状腺激素水平的波动相吻合。对黑斑蛙甲状腺系统的研究,可为日后使用黑斑蛙开展甲状腺干扰作用的研究提供基础。       

Influence of density and predators on metamorphic size in Rhinella schneideri tadpoles raised in mesocosm conditions

Amphibians exhibit extreme plasticity in the timing of metamorphosis, and several species respond to water availability, accelerating metamorphosis when their ponds dry. We analyzed the plasticity of the developmental response to water volume in Rhinella schneideri tadpoles. We raised tadpoles in mesocosm. Covariation between body size at metamorphosis and timing of development was positive. Nevertheless, the first approximately 53% of the metamorphoses finishing the cycle required between 34 and 56 days, and the covariation between body size at metamorphosis and timing of development was negative. For these tadpoles, the larval density and the presence of predators did not significantly affect their mass to metamorphosis. Nevertheless, predators affected time to metamorphosis. For the remainder of the tadpoles that reached metamorphosis at > 56 days, the relationship between body size at metamorphosis and timing of development was positive. For these tadpoles, larval density was important for mass at metamorphosis and presence of predators was also important for time to metamorphosis. Two dominant features were observed: (i) approximately 53% of metamorphs had morphological features similar to individuals developing in desiccating ponds, and (ii) the other individuals had morphological characteristics comparable to metamorphs developing in an unchanging environment.     

Hemps, a novel EGF-like protein, plays a central role in ascidian metamorphosis

All chordates share several characteristic features including a dorsal hollow neural tube, a notochord, a pharynx and an endostyle. Unlike other chordate taxa, ascidians have a biphasic life-history with two distinct body plans. During metamorphosis, the larval nerve cord and notochord degenerate and the pharyngeal gill slits and endostyle form. While ascidians, like other marine invertebrates, metamorphose in response to specific environmental cues, it remains unclear how these cues trigger metamorphosis. We have identified a novel gene (Hemps) which encodes a protein with a putative secretion signal sequence and four epidermal growth factor (EGF)-like repeats which is a key regulator of metamorphosis in the ascidian Herdmania curvata. Expression of Hemps increases markedly when the swimming tadpole larva becomes competent to undergo metamorphosis and then during the first 24 hours of metamorphosis. The Hemps protein is localised to the larval papillae and anterior epidermis of the larva in the region known to be required for metamorphosis. When the larva contacts an inductive cue the protein is released, spreading posteriorly and into the tunic as metamorphosis progresses. Metamorphosis is blocked by incubating larvae in anti-Hemps antibodies prior to the addition of the cue. Addition of recombinant Hemps protein to competent larvae induces metamorphosis in a concentration-dependent manner. A subgroup of genes are specifically induced during this process. These results demonstrate that the Hemps protein is a key regulator of ascidian metamorphosis and is distinct from previously described inducers of this process in terrestrial arthropods and aquatic vertebrates.     

Energetics of metamorphic climax in the pickerel frog (Lithobates palustris)

Anuran metamorphosis, the transition from aquatic larvae to terrestrial juveniles, is accompanied by significant morphological, physiological, and behavioral changes. Timing of metamorphosis and final size, which can influence adult fitness, may depend on sufficient energy accumulated during the larval period to support metamorphosis. However, only two species of anurans have been examined for energetic costs of metamorphosis, Rana tigrina and Anaxyrus terrestris. Based on these species, it has been hypothesized that differences in energy expenditure are related to duration of metamorphosis. To compare energetic costs of metamorphosis among species and examine this hypothesis, we quantified the total energy required for metamorphosis of Lithobates palustris tadpoles by measuring oxygen consumption rates over the duration of metamorphic climax using closed-circuit respirometry. Total energy costs for L. palustris were positively related to tadpole mass and duration of metamorphic climax. However, larger tadpoles completed metamorphosis more efficiently because they used proportionally less total energy for metamorphic climax than smaller counterparts. Costs were intermediate to R. tigrina, a larger species with similar metamorphic duration, and A. terrestris, a smaller species with shorter metamorphic climax. The results supported the hypothesis that amphibian species with more slowly developing tadpoles, such as ranids, require more absolute energy for metamorphosis in comparison to more rapidly developing species like bufonids.     

Active metabolism of thyroid hormone during metamorphosis of amphioxus

Thyroid hormones (THs), and more precisely the 3,3',5-triiodo-l-thyronine (T(3)) acetic derivative 3,3',5-triiodothyroacetic acid (TRIAC), have been shown to activate metamorphosis in amphioxus. However, it remains unknown whether TRIAC is endogenously synthesized in amphioxus and more generally whether an active TH metabolism is regulating metamorphosis. Here we show that amphioxus naturally produces TRIAC from its precursors T(3) and l-thyroxine (T(4)), supporting its possible role as the active TH in amphioxus larvae. In addition, we show that blocking TH production inhibits metamorphosis and that this effect is compensated by exogenous T(3), suggesting that a peak of TH production is important for advancement of proper metamorphosis. Moreover, several amphioxus genes encoding proteins previously proposed to be involved in the TH signaling pathway display expression profiles correlated with metamorphosis. In particular, thyroid hormone receptor (TR) and deiodinases gene expressions are either up- or down-regulated during metamorphosis and by TH treatments. Overall, these results suggest that an active TH metabolism controls metamorphosis in amphioxus, and that endogenous TH production and metabolism as well as TH-regulated metamorphosis are ancestral in the chordate lineage.     

Habitat deterioration promotes the evolution of direct development in metamorphosing species

Although metamorphosis is widespread in the animal kingdom, several species have evolved life-cycle modifications to avoid complete metamorphosis. Some species, for example, many salamanders and newts, have deleted the adult stage via a process called paedomorphosis. Others, for example, some frog species and marine invertebrates, no longer have a distinct larval stage and reach maturation via direct development. Here we study which ecological conditions can lead to the loss of metamorphosis via the evolution of direct development. To do so, we use size-structured consumer-resource models in conjunction with the adaptive-dynamics approach. In case the larval habitat deteriorates, individuals will produce larger offspring and in concert accelerate metamorphosis. Although this leads to the evolutionary transition from metamorphosis to direct development when the adult habitat is highly favorable, the population will go extinct in case the adult habitat does not provide sufficient food to escape metamorphosis. With a phylogenetic approach we furthermore show that among amphibians the transition of metamorphosis to direct development is indeed, in line with model predictions, conditional on and preceded by the evolution of larger egg sizes.     

GENETIC VARIATION FOR PHENOTYPIC PLASTICITY IN THE LARVAL LIFE HISTORY OF SPADEFOOT TOADS (SCAPHIOPUS COUCHII)

Phenotypic plasticity in life-history traits is common. The relationship between phenotype and environment, or reaction norm, associated with life-history plasticity can evolve by natural selection if there is genetic variation within a population for the reaction norm and if the traits involved affect fitness. As with other traits, selection on plasticity in a particular trait or in response to a particular environmental factor may be constrained by trade-offs with other traits that affect fitness. In this paper, I experimentally evaluated broad-sense genetic variation in the reaction norms of age and size at metamorphosis in response to two environmental factors, food level and temperature. Differences among full-sib families in one or both traits were evident in all treatments. However, variation among families in their responses to each treatment (genotype-environment interaction) resulted in variation among treatments in estimated heritabilities and genetic correlations. Age at metamorphosis was equally sensitive to temperature in all families, but size at metamorphosis was more sensitive to temperature in some families than in others. Size at metamorphosis was equally sensitive to food level in all families, but age at metamorphosis was sensitive to food in some families but not in others. At high temperature or low food, the genetic correlation between age and size at metamorphosis was positive, generating a potential trade-off between metamorphosing early to attain higher larval survival and metamorphosing later to achieve larger size. This trade-off extends across treatments: families with the largest average size at metamorphosis achieved larger size with the longest average and greatest plasticity in age at metamorphosis. Other families achieved shorter average larval periods by exhibiting greater plasticity in size at metamorphosis but had the smallest average size at metamorphosis. This trade-off may reflect an underlying functional constraint on the ability to respond optimally to all environments, resulting in persistent genetic variation in reaction norms.     

Metamorphosis of a scleractinian coral in response to microbial biofilms

Microorganisms have been reported to induce settlement and metamorphosis in a wide range of marine invertebrate species. However, the primary cue reported for metamorphosis of coral larvae is calcareous coralline algae (CCA). Herein we report the community structure of developing coral reef biofilms and the potential role they play in triggering the metamorphosis of a scleractinian coral. Two-week-old biofilms induced metamorphosis in less than 10% of larvae, whereas metamorphosis increased significantly on older biofilms, with a maximum of 41% occurring on 8-week-old microbial films. There was a significant influence of depth in 4- and 8-week biofilms, with greater levels of metamorphosis occurring in response to shallow-water communities. Importantly, larvae were found to settle and metamorphose in response to microbial biofilms lacking CCA from both shallow and deep treatments, indicating that microorganisms not associated with CCA may play a significant role in coral metamorphosis. A polyphasic approach consisting of scanning electron microscopy, fluorescence in situ hybridization (FISH), and denaturing gradient gel electrophoresis (DGGE) revealed that coral reef biofilms were comprised of complex bacterial and microalgal communities which were distinct at each depth and time. Principal-component analysis of FISH data showed that the Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Cytophaga-Flavobacterium of Bacteroidetes had the largest influence on overall community composition. A low abundance of Archaea was detected in almost all biofilms, providing the first report of Archaea associated with coral reef biofilms. No differences in the relative densities of each subdivision of Proteobacteria were observed between slides that induced larval metamorphosis and those that did not. Comparative cluster analysis of bacterial DGGE patterns also revealed that there were clear age and depth distinctions in biofilm community structure; however, no difference was detected in banding profiles between biofilms which induced larval metamorphosis and those where no metamorphosis occurred. This investigation demonstrates that complex microbial communities can induce coral metamorphosis in the absence of CCA.     

Differentiations of 5-HT and GAS cells in the digestive canals of Rana chensinensis tadpoles

In the current study, 5-nydroxytryptamine(5-HT) and gastrin(GAS) cells in the digestive canals of Rana chensinensis tadpoles at different developmental stages were investigated by immunohistochemistry. Results showed that the 5-HT cells were only detected in the duodenum before metamorphosis began, and were extensively distributed in the stomach, duodenum, small intestine, and rectum thereafter, with the highest counts found in the duodenum and rectum when metamorphosis was completed. The GAS cells were only distributed in the stomach and duodenum, and only rarely detected in the duodenum before metamorphosis began, but increased in the stomach during metamorphosis and showed zonal distribution in the gastric mucosa when metamorphosis was completed. Metamorphosis is a critical period for amphibians, during which structural and functional physiological adaptations are required to transition from aquatic to terrestrial environments. During metamorphosis, the differentiations of 5-HT cells in the gastrointestinal canals of tadpoles could facilitate mucus secretion regulation, improve digestive canal lubrication, and help watershortage food digestion in terrestrial environments. Conversely, GAS cell differentiations during metamorphosis might contribute to the digestive and absorptive function transition from herbivore to omnivore.     

Why grow up? A perspective on insect strategies to avoid metamorphosis

1. Insects with complete metamorphosis (holometaboly) are extremely successful, constituting over 60% of all described animal species. Complete metamorphosis confers significant advantages because it enables organisms to optimise life‐history components through temporal partitioning, and thereby to exploit multiple ecological niches. Yet holometaboly can also impose costs, and several lineages have evolved life cycle modifications to avoid complete metamorphosis. 2. In this review, we discuss different strategies that have evolved that result in the loss of complete metamorphosis (type I and type II paedomorphosis). In addition, the ecological pressures and developmental modifications that facilitate this avoidance are considered, as well as the importance of life cycle complexity in life‐history evolution. 3. Interestingly, only female holometabolous insects have entirely avoided complete metamorphosis, and it is always the ancestrally juvenile morphology that is retained. These findings point to a strong sex‐biased trade‐off between investment in reproduction and development. While the loss of complete metamorphosis in females has occurred independently on several occasions across holometabolous insects, only a small number of species possessing this ability have been described. 4. Thus, complete metamorphosis, which originated only once in insects, appears to have been almost fully retained. This indicates that significant modifications to the holometabolan metamorphic ground plan are highly constrained, and suggests that the transition to complete metamorphosis is evolutionarily irreversible.     

Metamorphosis of the feeding mechanism in tiger salamanders (Ambystoma tigrinum): the ontogeny of cranial muscle mass

Most previous research on metamorphosis of the musculoskeletal system in vertebrates has focused on the transformation of the skeleton. In this paper we focus on the transformation of the muscles of the head during metamorphosis in tiger salamanders ( Ambystoma tigrinum ) in order (1) to provide new data on changes in myology during ontogeny, and (2) to aid in interpreting previous data on the metamorphosis of function in the head of salamanders.
The physiological cross-sectional area of nine head muscles was calculated by measuring fibre angles, fibre lengths, and muscle mass in two samples of tiger salamanders obtained just before and just after metamorphosis. The major mouth-opening muscles (rectus cervicis and depressor mandibulae) exhibit a significant decrease in estimated maximum tetanic tension (MTT) across metamorphosis of about 36%. The jaw-closing muscles (adductor mandibulae internus and externus) and the head-lifting muscles (epaxials) also decrease in MTT but not significantly. The muscles associated with tongue projection during feeding on land (the subarcualis rectus I, geniohyoideus, interhyoideus and intermandibularis) all show a slight increase in MTT at metamorphosis.
Metamorphic transformation of feeding behaviour in Ambystoma tigrinum involves changes in performance, the design of skeletal elements, changes in muscle force-generating capability, and changes in hydrodynamic design from unidirectional flow in larvae to bidirectional flow during aquatic feeding after metamorphosis. Although muscle activity patterns during aquatic feeding do not change across metamorphosis, tongue-based terrestrial feeding involves a suite of novel muscle activity patterns, morphological characters acquired at metamorphosis, and a metamorphic increase in the masses of muscles important in tongue projection.     

Is Lamprey Metamorphosis Regulated by Thyroid Hormones?

Lampreys are one of just a few fishes which have a true (firstor first type) of metamorphosis in their life cycle. In thesea lamprey (Petromyzon marinus), spontaneous metamorphosisis initiated when the size (length and weight), condition factor,and lipid stores reach appropriate levels and coincide withthe postwinter rise in water temperature. The serum levels ofthe thyroid hormones, thyroxine (T₄) and triiodothyronine (T₃),drop dramatically at the onset of metamorphosis and metamorphosiscan be induced with treatment of animals with the goitrogen,KCIO₄, which also results in a decline in serum levels of thyroidhormones. The fact that thyroid hormone treatment can blockspontaneous and induced metamorphosis is support for the viewthat thyroid hormones, particularly T₃, operates like a juvenilehormone in lamprey metamorphosis; this view is counter to therole of thyroid hormones in metamorphosis of other vertebrates.The monodeiodinase pathways, whereby T₄ is converted to T₃ orto the biologically inactive reverse T₃, and even further degradationof T₃, may be a significant mechanism directing metamorphicchange. Lamprey metamorphosis is facultative in that it is initiatedor inhibited depending upon the coordination of a complex integrationof environmental, metabolic and hormonal cues. Thyroid hormonesdo not regulate lamprey metamorphosis in the sense observedin other vertebrate metamorphoses but they are important tothe developmental process. Some of the features of the involvementof thyroid hormones in lamprey metamorphosis may be relatedto the presence of the endostyle in larvae which in turn reflectsthe ancient origins of this vertebrate and perhaps the conservationof an ancient method of induction of metamorphosis. Some cluefor other factors which initiate lamprey metamorphosis may comethrough the examination of inducers of metamorphosis in lowerchordates     

The dorsal light reactions of larval and metamorphosing flatfish

Before metamorphosis, plaice and turbot show dorsal light reactions generally similar to those of other species, suggesting that their balance mechanisms are also similar, and that their utriculi are functional. The reactions are unusual in one respect in that larvae of both species tilt away from light incident from below the horizontal. In turbot, during metamorphosis, the null position (that taken up when illumination is from directly above) becomes gradually inclined to the right until the larva is horizontal at the end of metamorphosis. There is evidence that the null position of the static system is reset to an inclined position during this change. In plaice, during metamorphosis, the shift in null position is smaller, in the opposite direction, and at the end of metamorphosis there is a rapid change to a horizontal position. The null position of the static system does not appear to be reset. In plaice the influence of light on balance increases during metamorphosis, possibly due to it affecting the brain or the otolith organs. This is not seen in turbot, but light has a strong influence on the balance of this species both before and during metamorphosis. At the end of metamorphosis both plaice and turbot appear to utilize only static information from the sacculi and lagenae. Light could be seen to influence balance in swimming turbot, but not in plaice.     

Energy accumulation and amphibian metamorphosis

Summary Timing of metamorphosis and size at metamorphosis were examined for Hyla crucifer under two densities in the laboratory. Results agree with previously known relationships for ranids and bufonids: at higher densities developmental time is longer, mean size at metamorphosis is smaller, and both parameters have greater variance than at lower densities. Density also affects energy accumulation: under ad libitum feeding conditions, an average individual raised at a higher density is able to accumulate less energy before metamorphosis than an average individual raised at a lower density. The suggestion is made that, in addition to growth and differentiation rates, energy accumulation may be important in determining when amphibian metamorphosis occurs.     

Larval development and post-settlement metamorphosis of the barnacle Balanus albicostatus Pilsbry and the serpulid polychaete Pomatoleios kraussii Baird: Impact of a commonly used antifouling biocide,Irgarol 1051

Abstract

The impact of a commonly-used antifouling algicide, Irgarol 1051, on the larval development and post-settlement metamorphosis of the barnacle, Balanus albicostatus Pilsbry (Crustacea: Cirripedia), and the larval metamorphosis of a serpulid polycheate, Pomatoleios kraussii Baird, was evaluated. In the case of B. albicostatus, larval mortality increased with an increase in the concentration of Irgarol 1051, and there was a shift in the larval stage targeted from advanced instars to early instars. Nauplii that survived to the cyprid instar stage when reared in the presence of Irgarol 1051 showed prolonged instar and total naupliar duration when compared to the controls. The post-settlement metamorphosis of cyprids significantly varied with Irgarol concentration and also with biofilm age. One and 2-d-old untreated biofilms showed higher metamorphosis when compared to 5-d-old biofilms. However, when the biofilms that promoted cyprid metamorphosis were treated with Irgarol 1051 at low concentrations, metamorphosis rates decreased. Cyprids were prevented from metamorphosing completely by biofilms treated at the highest concentration of Irgarol 1051. Inhibition of metamorphosis was also observed in the case of competent polychaete larvae when exposed to Irgarol 1051 compared to those exposed to metamorphosis inducers such as 3-iso-butyl-1-methylxanthine (IBMX) and natural biofilms. Identification of the pathway(s) that caused the promotory biofilms to become toxic when exposed to Irgarol 1051 is discussed.