Timing Of Metamorphosis Research Articles

EFFECTS OF FEED ON THE SURVIVAL RATE, GROWTH RATE AND METAMORPHOSIS TIME OF THE WHITE LEG SHRIMP LARVAE (Litopenaeus vannamei Boone, 1931) AT ZOEA STAGE

The larvae of white leg shrimp (Litopenaeus vannamei Boone, 1931) at phase nauplii 6 fed with a stocking density of 150 pcs per litre and 3 formula including formula 1 with dry microalgae Spirulina + artemia as main ingredients, formula 2 with microalgae Thalasiosira weissflogii + artemia as main ingredients and formula 3 with microalgae Thalasiosira pseudonana + artemia as main ingredients. Experiments were carried out in 18 tanks fifty-litre composite at Quang Binh branch of C.P Vietnam Corporation. The study results show that, with formula 3, white leg shrimps larvae at zoea stage have the highest growth and survival rates (86.57 %), followed by formula 2 (76.03 %) and finally formula 1 (67.90 %). White leg shrimps larvae at zoea stage in formula 3 have the shortest metamorphosis time (78.61 hours), followed by formula 2 (83.25 hours) and the longest in formula 1 (94.10 hours).

Genetic accommodation via modified endocrine signalling explains phenotypic divergence among spadefoot toad species

Phenotypic differences among species may evolve through genetic accommodation, but mechanisms accounting for this process are poorly understood. Here we compare hormonal variation underlying differences in the timing of metamorphosis among three spadefoot toads with different larval periods and responsiveness to pond drying. We find that, in response to pond drying, Pelobates cultripes and Spea multiplicata accelerate metamorphosis, increase standard metabolic rate (SMR), and elevate whole-body content of thyroid hormone (the primary morphogen controlling metamorphosis) and corticosterone (a stress hormone acting synergistically with thyroid hormone to accelerate metamorphosis). In contrast, Scaphiopus couchii has the shortest larval period, highest whole-body thyroid hormone and corticosterone content, and highest SMR, and these trait values are least affected by pond drying among the three species. Our findings support that the atypically rapid and canalized development of S. couchii evolved by genetic accommodation of endocrine pathways controlling metamorphosis, showing how phenotypic plasticity within species may evolve into trait variation among species.

Atrazine feminizes sex ratio in Blanchard's cricket frogs (Acris blanchardi) at concentrations as low as 0.1 μg/L.

We exposed Blanchard's cricket frogs (Acris blanchardi) to ecologically relevant concentrations (0, 0.1, 1, and 10 μg/L) of a commercial formulation of atrazine throughout the larval period to determine effects on survival, somatic growth and development (time to metamorphosis and mass at metamorphosis), and gonadal development (sex ratio at metamorphosis and the prevalence of testicular ova in phenotypic males). We tested the following hypotheses: 1) atrazine feminizes the sex ratio, 2) atrazine increases the proportion of phenotypic males with testicular ova, and 3) atrazine differentially affects somatic growth (mass at metamorphosis) and development (time to metamorphosis) for males and females. Although the control sex ratio was male-biased, exposure to 0.1 and 10 μg/L atrazine feminized sex ratios, because these treatments produced 51 and 55% fewer males than the control, respectively. We did not observe testicular ova. Atrazine did not impact survival or metamorphosis, and we did not detect sexually dimorphic impacts on time to metamorphosis or mass at metamorphosis. However, males metamorphosed 2.3 d later than females, regardless of treatment. Sex biases in timing of metamorphosis are underexplored in anurans, but if prevalent, could have important implications for theory surrounding the impact of environmental factors on metamorphosis. Our data suggest that cricket frog sex ratios are sensitive to environmentally relevant concentrations of atrazine and that feminization in the field is likely. Environ Toxicol Chem 2018;37:427-435. © 2017 SETAC.

Severe developmental timing defects in the prothoracicotropic hormone (PTTH)-deficient silkworm, Bombyx mori

The insect neuropeptide prothoracicotropic hormone (PTTH) triggers the biosynthesis and release of the molting hormone ecdysone in the prothoracic gland (PG), thereby controlling the timing of molting and metamorphosis. Despite the well-documented physiological role of PTTH and its signaling pathway in the PG, it is not clear whether PTTH is an essential hormone for ecdysone biosynthesis and development. To address this question, we established and characterized a PTTH knockout line in the silkworm, Bombyx mori. We found that PTTH knockouts showed a severe developmental delay in both the larval and pupal stages. Larval phenotypes of PTTH knockouts can be classified into three major classes: (i) developmental arrest during the second larval instar, (ii) precocious metamorphosis after the fourth larval instar (one instar earlier in comparison to the control strain), and (iii) metamorphosis to normal-sized pupae after completing the five larval instar stages. In PTTH knockout larvae, peak levels of ecdysone titers in the hemolymph were dramatically reduced and the timing of peaks was delayed, suggesting that protracted larval development is a result of the reduced and delayed synthesis of ecdysone in the PG. Despite these defects, low basal levels of ecdysone were maintained in PTTH knockout larvae, suggesting that the primary role of PTTH is to upregulate ecdysone biosynthesis in the PG during molting stages, and low basal levels of ecdysone can be maintained in the absence of PTTH. We also found that mRNA levels of genes involved in ecdysone biosynthesis and ecdysteroid signaling pathways were significantly reduced in PTTH knockouts. Our results provide genetic evidence that PTTH is not essential for development, but is required to coordinate growth and developmental timing.

Dynamics of growth-based survival mechanisms in Japanese anchovy (Engraulis japonicus) larvae

Three growth-based survival mechanisms were tested for multiple cohorts of Japanese anchovy (Engraulis japonicus) larvae in Sagami Bay. Through otolith microstructure analysis, growth trajectories and histories of samples of the survivors were compared with those of the original populations to examine size- and growth-selective mortality to test the “bigger is better” and “growth-selective predation” mechanisms, respectively. The effects of growth rates on the timing of metamorphosis were examined to test the “stage duration” mechanism. The bigger is better and growth-selective predation mechanisms were detected to be effective in two and six of eight seasonal cohorts, respectively. Results contrary to the bigger is better and growth-selective predation mechanisms were obtained from three and two of eight cohorts, respectively. The stage duration mechanism was evaluated to be effective for both of two cohorts that were testable. Overall, none of the three mechanisms were universally appropriate for all of the cohorts. The relative contributions of the three mechanisms were dynamic, although the growth-selective predation mechanism was identified to be the major one in anchovy larvae in the study site.

Global gene expression analysis provides insight into local adaptation to geothermal streams in tadpoles of the Andean toad Rhinella spinulosa

The anuran Rhinella spinulosa is distributed along the Andes Range at altitudes that undergo wide daily and seasonal variation in temperature. One of the populations inhabits geothermal streams, a stable environment that influences life history traits such as the timing of metamorphosis. To investigate whether this population has undergone local adaptation to this unique habitat, we carried out transcriptome analyses in animals from two localities in two developmental stages (prometamorphic and metamorphic) and exposed them to two temperatures (20 and 25 °C). RNA-Seq, de novo assembly and annotation defined a transcriptome revealing 194,469 high quality SNPs, with 1,507 genes under positive selection. Comparisons among the experimental conditions yielded 1,593 differentially expressed genes. A bioinformatics search for candidates revealed a total of 70 genes that are highly likely to be implicated in the adaptive response of the population living in a stable environment, compared to those living in an environment with variable temperatures. Most importantly, the population inhabiting the geothermal environment showed decreased transcriptional plasticity and reduced genetic variation compared to its counterpart from the non-stable environment. This analysis will help to advance the understanding of the molecular mechanisms that account for the local adaptation to geothermal streams in anurans.

Implantation assays using the integument of early stage Bombyx larvae: Insights into the mechanisms underlying the acquisition of competence for metamorphosis

It is widely accepted that the anti-metamorphic action of insect juvenile hormones (JHs) is required to inhibit larval-pupal metamorphosis. However, recent studies using RNAi or knockout techniques reveal that larval status may be maintained independently of JHs during the early larval stages. To investigate why larvae of very early instars do not have competence to metamorphose and how they acquire this competence through larval development, we revisited the classic experiments of Piepho (ca. 1930s) and performed implantation assays using the integument of very young larvae of the silkworm, Bombyx mori. Here, we demonstrate that when the integuments of neonate larvae or newly molted second instar larvae are implanted into last instar host larvae, they are able to directly produce pupal cuticle at the time of pupal metamorphosis of the host. To investigate whether the pupal commitment of implants from the neonate first instar larvae is repressed by JHs, the integuments of Met1 knockout larvae lacking a functional JH receptor were implanted into penultimate instar larvae. We found that the implants of Met1 knockout neonate larvae produced patched pupal cuticles after the host larval molt, whereas those of the wild-type strain produced only larval cuticle without any trace of pupal cuticle. Taken together, our results suggest that the epidermis of very early instar larvae can be pupally committed when provided with unidentified blood-borne factor(s) present in final-instar larvae, and that JHs can block the action of that factor(s) to prolong the feeding period until larvae attain a size appropriate for metamorphosis.

Weather, hydroregime, and breeding effort influence juvenile recruitment of anurans: implications for climate change

AbstractAmphibians that primarily breed in ephemeral wetlands are especially vulnerable to climate change because they rely on rainfall or temperature to initiate breeding and create suitable hydroregimes (water duration, timing, frequency, depth) for reproductive success. Hydroregime effects on reproductive success are likely to differ among species because of differences in reproductive strategies: the length and timing of breeding period, rate of larval development, and timing of metamorphosis. We applied an information‐theoretic approach to 22 consecutive years of continuous amphibian trapping data at eight ephemeral wetlands to test hypotheses regarding environmental (hydroregime, weather) and biological (adult breeding effort) factors affecting juvenile recruitment (JR) by six focal species representing four reproductive strategies. We hypothesized that (1) JR by species with similar reproductive strategies would be influenced by similar variables; (2) JR would be higher for all species when models encompassed the maximum time span of potential tadpole occurrence and development; and (3) JR rates within individual wetlands and breeding cycles would correlate most closely between species with similar breeding strategies. The best model for all focal species (except Scaphiopus holbrookii) encompassed the maximum time span and indicated that ≥1 hydroregime variable, total precipitation, or both were important drivers of reproductive success; average air temperature was not. Continuous hydroperiod through peak juvenile emigration was an important predictor of JR for species with prolonged breeding periods, slow larval development, and a “fixed” late spring start date for juvenile emigration (regardless of when oviposition occurred, or cohort age; Lithobates capito, Lithobates sphenocephalus), but not for species with rapid larval development and continual emigration as cohorts complete metamorphosis (Anaxyrus terrestris, Anaxyrus quercicus, Gastrophryne carolinensis, S. holbrookii). Total rainfall was positively associated with recruitment for most species; depth characteristics affected species differently. Annual JR was positively correlated among species with similar reproductive strategies. Our results indicate that weather and hydroregime characteristics interact with reproductive strategies that differ among amphibian species and influence reproductive plasticity, opportunity, and success. Effects of altered weather patterns associated with climate change on amphibian reproductive success may correspond more closely among species having similar reproductive strategies, with critical implications for population trends and assemblages.

Effect of triclosan on anuran development and growth in a larval amphibian growth and development assay.

A larval amphibian growth and development assay was performed to evaluate the potential effects of environmentally-relevant concentrations of triclosan (TCS) on amphibian development and growth. Xenopus laevis were exposed to TCS 0.0 (control), 6.3, 12.5 and 25.0 μg l-1 (estimated maximum tolerable concentration) until 10 weeks post-metamorphosis. At median metamorphosis time (Nieuwkoop and Faber stage 62), five larvae per replicate were collected for snout-vent length, hind limb length and body weight measurements, and histopathological examination of thyroid glands. Endpoints evaluated at test termination were based on draft guidance (USEPA, ) and included: survival; snout-vent length; body weight; gender; nuptial pad development (males); and liver, kidney, gonad and gonadal ducts histopathology. Exposure to TCS did not decrease survival, induce general signs of toxicity, affect median metamorphosis time or alter sex ratios. Exposure to TCS 12.5 and 25 μg l-1 increased growth during the metamorphic stages relative to the control, but did not influence growth during the post-metamorphic phase. Overall, several statistically significant findings were found in larvae exposed to TCS, such as a decrease in the prevalence of stage 3 Müllerian ducts in the anterior trunk sections of TCS 25.0 μg l-1 dose group females as compared to controls; most were not considered toxicologically relevant. Copyright © 2017 John Wiley & Sons, Ltd.

Thyroid Hormone Receptor α Controls Developmental Timing and Regulates the Rate and Coordination of Tissue-Specific Metamorphosis in Xenopus tropicalis.

Thyroid hormone (T3) receptors (TRs) mediate the effects of T3 on organ metabolism and animal development. There are two TR genes, TRα and TRβ, in all vertebrates. During animal development, TRα expression is activated earlier than zygotic T3 synthesis and secretion into the plasma, implicating a developmental role of TRα both in the presence and absence of T3. Using T3-dependent amphibian metamorphosis as a model, we previously proposed a dual-function model for TRs, in particular TRα, during development. That is, unliganded TR represses the expression of T3-inducible genes during premetamorphosis to ensure proper animal growth and prevent premature metamorphosis, whereas during metamorphosis, liganded TR activates target gene transcription to promote the transformation of the tadpole into a frog. To determine if TRα has such a dual function, we generated homozygous TRα-knockout animal lines. We show that, indeed, TRα knockout affects both premetamorphic animal development and metamorphosis. Surprisingly, we observed that TRα is not essential for amphibian metamorphosis, given that homozygous knockout animals complete metamorphosis within a similar time period after fertilization as their wild-type siblings. On the other hand, the timing of metamorphosis for different organs is altered by the knockout; limb metamorphosis occurs earlier, whereas intestinal metamorphosis is completed later than in wild-type siblings. Thus, our studies have demonstrated a critical role of endogenous TRα, not only in regulating both the timing and rate of metamorphosis, but also in coordinating temporal metamorphosis of different organs.

The potential effects of pre-settlement processes on post-settlement growth and survival of juvenile northern rock sole (Lepidopsetta polyxystra) in Gulf of Alaska nursery habitats

Early life history traits in marine fish such as growth, size, and timing of life history transitions often vary in response to environmental conditions. Identifying the potential effects of trait variation across life history stages is critical to understanding growth, recruitment, and survival. Juvenile northern rock sole (Lepidopsetta polyxystra) were collected (2005, 2007, 2009–2011) from two coastal nurseries in the Gulf of Alaska during the early post-settlement period (July–August) to examine variation in early life history traits in relation to water temperature and juvenile densities in nurseries as well as to evaluate the potential for carry-over effects. Size-at-hatch, larval growth, metamorphosis size and timing, and post-metamorphic and recent growth of juveniles were quantified using otolith structural analysis and compared across years and sites. Additionally, traits of fish caught in July and August were compared for evidence of selective mortality. Post-metamorphic and recent growth were related to temperatures in nurseries as well as temperatures during the larval period, indicating a direct influence of concurrent nursery temperatures and a potential indirect effect of thermal conditions experienced by larvae. Correlations between metamorphic traits and fish size at capture demonstrated that interannual variation in size persisted across life history stages regardless of post-settlement growth patterns. No evidence of density-dependent growth or growth-selective mortality were detected during the early post-settlement period; however, differences in hatch size and metamorphosis timing between fish collected in July and August indicate a selective loss of individuals although the pattern varied across years. Overall, variation in size acquired early in life and temperature effects on the phenology of metamorphosis may influence the direction of selection and survival of northern rock sole.

How Metamorphosis Is Different in Plethodontids: Larval Life History Perspectives on Life-Cycle Evolution.

Plethodontid salamanders exhibit biphasic, larval form paedomorphic, and direct developing life cycles. This diversity of developmental strategies exceeds that of any other family of terrestrial vertebrate. Here we compare patterns of larval development among the three divergent lineages of biphasic plethodontids and other salamanders. We discuss how patterns of life-cycle evolution and larval ecology might have produced a wide array of larval life histories. Compared with many other salamanders, most larval plethodontids have relatively slow growth rates and sometimes exceptionally long larval periods (up to 60 mo). Recent phylogenetic analyses of life-cycle evolution indicate that ancestral plethodontids were likely direct developers. If true, then biphasic and paedomorphic lineages might have been independently derived through different developmental mechanisms. Furthermore, biphasic plethodontids largely colonized stream habitats, which tend to have lower productivity than seasonally ephemeral ponds. Consistent with this, plethodontid larvae grow very slowly, and metamorphic timing does not appear to be strongly affected by growth history. On the basis of this, we speculate that feeding schedules and stress hormones might play a comparatively reduced role in governing the timing of metamorphosis of stream-dwelling salamanders, particularly plethodontids.

Mechanisms of Density-dependent Growth and Survival in Tadpoles of Fowler's Toad,Anaxyrus fowleri: Volume vs. Abundance

Density-dependent growth has ordinarily been studied in aquatic ecosystems by varying the abundance of animals in mesocosms of equal volume. Aside from the unequal sample sizes involved with using this abundance-limited method, confounding factors potentially associated with levels of social interactions may also be introduced. The alternative, volume-limited method, i.e., varying the volume while maintaining equal numbers of animals, can provide a test for the presence of potentially confounding factors. Using tadpoles of Fowler's Toad, Anaxyrus fowleri, we examined the effect of density on growth rate, timing of metamorphosis, and size at metamorphosis in both abundance-limited and volume-limited experiments. We found no difference in tadpole growth rate or timing of metamorphosis between these two methods, but the metamorphs emerging from abundance-limited low density treatments were significantly smaller when compared to those in volume-limited low density treatments. Because toad tadpoles may natural...

Influence of Temperature on the Thyroidogenic Effects of Diuron and Its Metabolite 3,4-DCA in Tadpoles of the American Bullfrog (Lithobates catesbeianus).

Temperature is a key variable affecting the timing of amphibian metamorphosis from tadpoles to tetrapods, through the production and subsequent function of thyroid hormones (TH). Thyroid function can be impaired by environmental contaminants as well as temperature. Tadpoles can experience large temperature fluctuations in their habitats and many species are distributed in areas that may be impacted by agriculture. Diuron is a widely used herbicide detected in freshwater ecosystems and may impact endocrine function in aquatic organisms. We evaluated the influence of temperature (28 and 34 °C) on the action of diuron and its metabolite 3,4-dichloroaniline (3,4-DCA) on thyroid function and metamorphosis in tadpoles of Lithobates catesbeianus. Exposure to both compounds induced more pronounced changes in gene expression and plasma 3,3',5-triiodothyronine (T3) concentrations in tadpoles treated at higher temperature. T3 concentrations were increased in tadpoles exposed to 200 ng/L of diuron at 34 °C and an acceleration of metamorphosis was observed for the same group. Transcriptomic responses included alteration of thyroid hormone induced bZip protein (thibz), deiodinases (dio2, dio3), thyroid receptors (trα, trβ) and Krüppel-like factor 9 (klf9), suggesting regulation by temperature on TH-gene expression. These results suggest that environmental temperature should be considered in risk assessments of environmental contaminants for amphibian species.

Lethal and sublethal effects of a methomyl-based insecticide in Hoplobatrachus rugulosus.

The aim of this study was to investigate the lethal and sublethal toxicity of a methomyl-based insecticide in Hoplobatrachus rugulosus, as methomyl-based insecticides are applied in massive amounts and agrochemicals have effects on the decline in amphibian populations. To evaluate the toxic effects of methomyl from agricultural application, a methomyl-based insecticide containing 40% methomyl was selected. The median lethal concentration of 96 hours of methomyl exposure was 8.69 ppm for H. rugulosus tadpoles. The lethal concentration also produced severe histological damage in the livers and kidneys of the exposed tadpoles. The sublethal concentration used for methomyl was 144 ppb during the metamorphosis period. It was found that the sublethal concentration of the methomyl compound could decrease growth, metamorphosis time, and size, disturb biochemical parameters, and produce histological damage. In livers, methomyl effects increased oxidative stress and dramatically decreased the glycogen level of the treated froglets. Mononuclear infiltration, blood congestion, amorphous substances, and hepatocytes vacuolization were observed throughout liver tissue. The methomyl-based insecticide also increased oxidative stress and decreased nitric oxide levels in the kidneys of the exposed froglets. Renal tissue damage including blood congestion, amorphous substances, and Bowman’s capsule spaces reduction were found in the methomyl exposure group. The methomyl compound also produced vacuoles in various stages of oocytes, but no histological damage was found in testicular tissue. Our results indicated strong toxic effects of the methomyl-based insecticide on H. rugulosus, a broadly tolerant anuran.

Cohort patterns in Rana tadpole metamorphosis

AbstractThis study explores the cohort patterns in a size versus time plot of individual metamorphs of Rana tadpoles. This meta‐study was based on a number of studies where any of three species of Rana tadpoles were raised in tanks under various treatments (variation in source pond, tadpole density, drying regime, hatching date, food level and competitors). A frequent pattern was found where there was an initial decrease in metamorph size as more slowly developing tadpoles metamorphosed. This negative trend sometimes changed to positive as later metamorphs appeared. The later metamorphs were often as large, or even larger, than those first to metamorphose, giving the plot a U‐shaped pattern. Also, a number of field studies of two of the Rana species’ were analysed for the same aspect. The U‐shaped pattern was much rarer but it did occur. A positive linear slope, rather than a negative one, was also found occasionally. A large number of factors affect the growth and development rates in tadpoles, adaptively and non‐adaptively, and thus determine their size at the time of metamorphosis. How these can contribute to the found metamorphosis patterns is discussed.

Insights into How Longicorn Beetle Larvae Determine the Timing of Metamorphosis: Starvation-Induced Mechanism Revisited.

Larvae of holometabolous insects must determine the timing of their metamorphosis. How they determine this timing has only been studied in detail for a few insect species. In a few species of Coleoptera, starvation is known to be a cue for metamorphosis, leading to the formation of smaller adults (starvation-induced pupation, SiP). We investigated the occurrence of SiP in the beetle Psacothea hilaris. When P. hilaris larvae were starved late in the feeding phase of the last (5th) instar, they exhibited typical SiP characterized by constancy of the time from food deprivation to pupation (TTP) irrespective of the body weight upon food deprivation or the length of prior feeding. In contrast, when larvae were starved early in the feeding phase, TTP decreased by roughly 1 day as the feeding became 1 day longer. The change in the response to starvation was estimated to occur on day 5.9 in the last instar. A series of refeeding experiments suggested that whereas SiP occurred readily in the larvae starved in the late feeding phase, activation of SiP was suspended until day 5.9 in the larvae starved early in the feeding phase. When P. hilaris larvae were fed continuously, they eventually ceased feeding spontaneously and pupated. The time length between spontaneous cessation of feeding and pupation was approximately equal to the TTP in SiP. This suggests that the same mechanism was activated by food deprivation in the late feeding phase and by spontaneous cessation of ad libitum feeding.

BDE-47 causes developmental retardation with down-regulated expression profiles of ecdysteroid signaling pathway-involved nuclear receptor (NR) genes in the copepod Tigriopus japonicus

2,2′,4,4′-Tetrabromodiphenyl ether (BDE-47) is a persistent organic pollutant (POP) in marine environments. Despite its adverse effects (e.g. developmental retardation) in ecdysozoa, the effects of BDE-47 on transcription of ecdysteroid signaling pathway-involved-nuclear receptor (NR) genes and metamorphosis-related genes have not been examined in copepods. To examine the deleterious effect of BDE-47 on copepod molting and metamorphosis, BDE-47 was exposed to the harpacticoid copepod Tigriopus japonicus, followed by monitoring developmental retardation and transcriptional alteration of NR genes. The developmental rate was significantly inhibited (P<0.05) in response to BDE-47 and the agricultural insecticide gamma-hexachlorocyclohexane. Conversely, the ecdysteroid agonist ponasterone A (PoA) led to decreased molting and metamorphosis time (P<0.05) from the nauplius stage to the adult stage. In particular, expression profiles of all NR genes were the highest at naupliar stages 5–6 except for SVP, FTZ-F1, and HR96 genes. Nuclear receptor USP, HR96, and FTZ-F1 genes also showed significant sex differences (P<0.05) in gene expression levels over different developmental stages, indicating that these genes may be involved in vitellogenesis. NR gene expression patterns showed significant decreases (P<0.05) in response to BDE-47 exposure, implying that molting and metamorphosis retardation is likely associated with NR gene expression. In summary, BDE-47 leads to molting and metamorphosis retardation and suppresses transcription of NR genes. This information will be helpful in understanding the molting and metamorphosis delay mechanism in response to BDE-47 exposure.

An Integrative Endocrine Model for the Evolution of Developmental Timing and Life History of Plethodontids and Other Salamanders

In recent years, many molecular endocrine mechanisms that regulate tissue morphogenesis have been detailed in laboratory amphibian models. However, most of these pathways have not been examined across more closely related species to understand how deviations in endocrine pathways may have contributed to amphibian diversification. The timing of metamorphosis and maturation vary extensively across plethodontid salamanders (including direct developing, biphasic, and paedomorphic species), making them an ideal system for analyzing the evolution of endocrine mechanisms in a phylogenetic context. Recent phylogenetic-based reconstructions concluded that ancestral plethodontids were likely direct developers, and free-living larval periods were independently derived multiple times within this family. Furthermore, within one clade (Spelerpini) there have been multiple independent transitions from biphasic (metamorphic) to paedomorphic developmental modes. This inspires the question: What endocrine/developmental mec...

Overcrowding-mediated stress alters cell proliferation in key neuroendocrine areas during larval development inRhinella arenarum

Exposure to adverse environmental conditions can elicit a stress response, which results in an increase in endogenous corticosterone levels. In early life stages, it has been thoroughly demonstrated that amphibian larval growth and development is altered as a consequence of chronic stress by interfering with the metamorphic process, however, the underlying mechanisms involved have only been partially disentangled. We examined the effect of intraspecific competition on corticosterone levels during larval development of the toad Rhinella arenarum and its ultimate effects on cell proliferation in particular brain areas as well as the pituitary gland. While overcrowding altered the number of proliferating cells in the pituitary gland, hypothalamus, and third ventricle of the brain, no differences were observed in areas which are less associated with neuroendocrine processes, such as the first ventricle of the brain. Apoptosis was increased in hypothalamic regions but not in the pituitary. With regards to pituitary cell populations, thyrotrophs but not somatoatrophs and corticotrophs showed a decrease in the cell number in overcrowded larvae. Our study shows that alterations in growth and development, produced by stress, results from an imbalance in the neuroendocrine systems implicated in orchestrating the timing of metamorphosis.