An integrative approach identifies developmental sequence heterochronies in freshwater basommatophoran snails

Adopting an integrative approach to the study of sequence heterochrony, we compared the timing of developmental events encompassing a mixture of developmental stages and functional traits in the embryos of 12 species of basommatophoran snails in an explicit phylogenetic framework. PARSIMOV analysis demonstrated clear functional heterochronies associated both with basal branches within the phylogeny and with terminal speciation events. A consensus of changes inferred under both accelerated transformation and delayed transformation optimizations identified four heterochronies where the direction of movement was known plus six twin heterochronies where the relative movements of the two events could not be assigned. On average, 0.5 and 0.58 events were inferred to have changed their position in the developmental sequence on internal and terminal branches of the phylogeny, respectively; these values are comparable with frequencies of sequence heterochrony reported in mammals. Directional heterochronies such as the early occurrence of body flexing in relation to the ontogeny of the eye spots, heart beat, and free swimming events occurred convergently and/or at different levels (i.e., familial, generic, and species) within the phylogeny. Such a functional approach to the study of developmental sequences has highlighted the possibility that heterochrony may have played a prominent role in the evolution of this group of invertebrates.     

Conserved relative timing of cranial ossification patterns in early mammalian evolution

We analyzed a comprehensive data set of ossification sequences including seven marsupial, 13 placental and seven sauropsid species. Data are provided for the first time for two major mammalian clades, Chiroptera and Soricidae, and for two rodent species; the published sequences of three species were improved with additional sampling. The relative timing of the onset of ossification in 17 cranial elements was recorded, resulting in 136 event pairs, which were treated as characters for each species. Half of these characters are constant across all taxa, 30% are variable but phylogenetically uninformative, and 19% potentially deliver diagnostic features for clades of two or more taxa. Using the conservative estimate of heterochronic changes provided by the program Parsimov, only a few heterochronies were found to diagnose mammals, marsupials, or placentals. A later onset of ossification of the pterygoid with respect to six other cranial bones characterizes therian mammals. This result may relate to the relatively small size of this bone in this clade. One change in relative onset of ossification is hypothesized as a potential human autapomorphy in the context of the sampling made: the earlier onset of the ossification of the periotic with respect to the lacrimal and to three basicranial bones. Using the standard error of scaled ranks across all species as a measure of each element's lability in developmental timing, we found that ossification of early, middle, and late events are similarly labile, with basicranial traits the most labile in timing of onset of ossification. Despite marsupials and placental mammals diverging at least 130 Ma, few heterochronic shifts in cranial ossification diagnose these clades.     

Inferring the past to predict the future: climate modelling predictions and phylogeography for the freshwater gastropod Radix balthica (Pulmonata, Basommatophora)

Understanding the impact of past climatic events on species may facilitate predictions of how species will respond to future climate change. To this end, we sampled populations of the common pond snail Radix balthica over the entire species range (northwestern Europe). Using a recently developed analytical framework that employs ecological niche modelling to obtain hypotheses that are subsequently tested with statistical phylogeography, we inferred the range dynamics of R. balthica over time. A Maxent modelling for present-day conditions was performed to infer the climate envelope for the species, and the modelled niche was used to hindcast climatically suitable range at the last glacial maximum (LGM) c . 21 000 years ago. Ecological niche modelling predicted two suitable areas at the LGM within the present species range. Phylogeographic model selection on a COI mitochondrial DNA data set confirmed that R. balthica most likely spread from these two disjunct refuges after the LGM. The match observed between the potential range of the species at the LGM given its present climatic requirements and the phylogeographically inferred refugial areas was a clear argument in favour of niche conservatism in R. balthica , thus allowing to predict the future range. The subsequent projection of the potential range under a global change scenario predicts a moderate pole-ward shift of the northern range limits, but a dramatic loss of areas currently occupied in France, western Great Britain and southern Germany.     

Parent–offspring similarity in the timing of developmental events: an origin of heterochrony?

Understanding the link between ontogeny (development) and phylogeny (evolution) remains a key aim of biology. Heterochrony, the altered timing of developmental events between ancestors and descendants, could be such a link although the processes responsible for producing heterochrony, widely viewed as an interspecific phenomenon, are still unclear. However, intraspecific variation in developmental event timing, if heritable, could provide the raw material from which heterochronies originate. To date, however, heritable developmental event timing has not been demonstrated, although recent work did suggest a genetic basis for intraspecific differences in event timing in the embryonic development of the pond snail, Radix balthica. Consequently, here we used high-resolution (temporal and spatial) imaging of the entire embryonic development of R. balthica to perform a parent–offspring comparison of the timing of twelve, physiological and morphological developmental events. Between-parent differences in the timing of all events were good predictors of such timing differences between their offspring, and heritability was demonstrated for two of these events (foot attachment and crawling). Such heritable intraspecific variation in developmental event timing could be the raw material for speciation events, providing a fundamental link between ontogeny and phylogeny, via heterochrony.     

Highly polymorphic microsatellite markers for Radix balthica (Linnaeus 1758)

We present data for eight polymorphic microsatellite markers isolated from a microsatellite-enriched DNA library for the freshwater snail Radix balthica. Three of them were specific for R. balthica while five also amplified polymorphic products in two congeneric species. Test application on populations from all over the species range has shown that these loci are highly informative for analysing population structure and estimating migration rates. Observed deviations from Hardy-Weinberg equilibrium are attributed to a mixed mating system.     

A genetic basis for intraspecific differences in developmental timing?

Heterochrony, altered developmental timing between ancestors and their descendents, has been proposed as a pervasive evolutionary feature and recent analytical approaches have confirmed its existence as an evolutionary pattern. Yet, the mechanistic basis for heterochrony remains unclear and, in particular, whether intraspecific variation in the timing of developmental events generates, or has the potential to generate, future between‐species differences. Here we make a key step in linking heterochrony at the inter‐ and intraspecific level by reporting an association between interindividual variation in both the absolute and relative timing (position within the sequence of developmental events) of key embryonic developmental events and genetic distance for the pond snail, Radix balthica. We report significant differences in the genetic distance of individuals exhibiting different levels of dissimilarity in their absolute and relative timing of developmental events such as spinning activity, eyespot formation, heart ontogeny, and hatching. This relationship between genetic and developmental dissimilarity is consistent with there being a genetic basis for variation in developmental timing and so suggests that intraspecific heterochrony could provide the raw material for natural selection to produce speciation.     

Phenotypically plastic responses of green frog embryos to conflicting predation risk

Predators have been shown to alter the timing of switch points between life history stages, but few studies have addressed switch point plasticity in prey exposed simultaneously to conflicting predation pressure. We tested hatching responses of green frog (Rana clamitans) embryos subject to perceived predation risk from chemical cues released by two stage-specific predators, predicting that these predators would elicit: (1) directional hatching responses when presented independently, and (2) intermediate phenotypic responses when presented simultaneously. R. clamitans embryos in outdoor exclosures were exposed to cues from an egg predator (freshwater leeches; Nephelopsis obscura), a larval predator (dragonfly nymphs, Aeschna canadensis), and both predators in a 2 × 2 factorial experiment, and changes in hatchling size, hatchling developmental stage, and hatching time were compared to those for control embryos. Leeches alone induced embryos to hatch at a smaller size and an earlier developmental stage than controls, while dragonfly nymphs elicited a delay in egg hatching time that was associated with larger size and later developmental stage at hatching. Embryos failed to respond to simultaneous exposure to both predators, implying that responses to each occurred concurrently and were therefore dampened. Our results indicate that prey under threat from conflicting predators may manifest intermediate defensive phenotypes. Such intermediate responses may result in elevated rates of prey mortality with possible consequences at the population level.     

Carry-over effects of ultraviolet-B radiation on larval fitness in Rana temporaria

A number of studies have failed to find evidence for negative effects of ultraviolet-B radiation (UVBR) on amphibian early-embryonic performance, leading to the conclusions, first, that the embryonic stages of many species are tolerant to UVBR, and second, that the increased amount of UVBR reaching the Earth's surface is not likely to have any direct negative effects on many amphibian populations. However, possible carry-over effects of exposure to UVBR in the embryonic stages to the larval stages have received less attention. We studied the effects of UVBR experienced during the embryonic stages (age less than 11 days) on the later performance (age 11-75 days) of common frog, Rana temporaria, larvae. In a factorial laboratory experiment, newly fertilized embryos were divided into three different UVBR treatments (no UVBR (control), 1.25 kJm(-2) (normal) and 1.58 kJm(-2) (26% enhanced)), after which the individual larvae were raised until metamorphosis in the absence of UVBR. No effects of UVBR on embryonic survival rates, frequency of developmental anomalies or hatchling size were found, corroborating the earlier results indicating that R. temporaria embryos are tolerant to UVBR. However, analyses of larval performance revealed that larvae exposed to enhanced levels of UVBR as embryos suffered from an increased frequency of developmental anomalies and metamorphosed later and at a smaller size than larvae that had been protected from UVBR as embryos. These results suggest, in contrast to the earlier studies, that UVBR has direct negative effects on R. temporaria embryos, but these effects are expressed mostly or only during the later life stages. To this end, our results support the contention that carry-over effects from one life stage to another may be an important source of phenotypic variation in fitness.     

Variance in developmental event timing is greatest at low biological levels: implications for heterochrony

Intraspecific variation in developmental event timing is common and may be the raw material from which heterochronies (altered timing of developmental events between ancestors and descendants) arise. However, our understanding of how variance in intraspecific developmental event timing is distributed across different hierarchical, biological levels is poor, despite recent evidence suggesting a genetic basis for such inter‐individual differences. In the present study, we used high (temporal and spatial) resolution bio‐imaging of the entire embryonic development of the pond snail, Radix balthica, to investigate how variance was partitioned between the biological levels of interpopulation, inter‐egg mass and individual, with respect to the relative timing of four key developmental events (four‐cell division, a discrete heart beat, capsule rupture, and hatching), as well as egg volume (an index of maternal investment), hatchling size and shape (to compare embryonic with post‐hatch variance). We found that the timing of all but one (four cell division) of the developmental events, together with all measures of hatchling size and shape, had most variance partitioned at the individual level; variance at the interpopulation level was surprisingly low despite sampling populations from across the geographical range of R. balthica. These patterns highlight the importance of studying the distribution of variance in developmental event timing with the aim of understanding how it might drive organismal ecology and evolution. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 581–590.     

Polymorphism in developmental timing: intraspecific heterochrony in a Lake Victoria cichlid

SUMMARY Biologists measure developmental time by dividing development into arbitrary time blocks called "stages." This is a reasonable approach, provided that developmental timing is precisely controlled within a species. However, the degree of this precision is unknown. This is unfortunate because precision in developmental timing at the population level is a central issue to the whole research program of heterochrony. To examine this issue, we apply Ontogenetic Sequence Analysis to 261 embryos of the Lake Victoria cichlid Haplochromis piceatus . The result of our analysis can be mapped as a complex web of 26,880 equally parsimonious developmental sequences. This topology reflects timing polymorphism (intraspecific heterochrony) among embryos of this species. Because of this timing polymorphism, it is not possible to define discrete "stages" in this cichlid (although there is sufficient sequence signal to assess the maturity of embryos). More generally, we show that sequence polymorphism creates uncertainty about how a given embryo will develop implying that the mechanisms controlling developmental timing in embryos lack precision. For this reason, it is imperative to consider patterns of embryonic variability when measuring developmental time.     

Early exposure to a bacterial endotoxin advances the onset of moult in the European starling

In animals, events occurring early in life can have profound effects on subsequent life‐history events. Early developmental stresses often produce negative long‐lasting impacts, although positive effects of mild stressors have also been documented. Most studies of birds have investigated the effects of events occurring at early developmental stages on the timing of migration or reproduction, but little is known on the long‐term effects of these early events on moulting and plumage quality. We exposed European starling Sturnus vulgaris nestlings to an immune challenge to assess the effects of a developmental stress on the timing of the first (post‐juvenile) and second (post‐breeding) complete annual moult, the length of the flight feathers, and the length and colouration of ornamental throat feathers. The nestlings were transferred to indoor aviaries before fledgling and kept in captivity until the end of post‐breeding moult. Individuals treated with Escherichia coli lypopolysaccharide (LPS) started both moult cycles earlier compared to control siblings. Moult duration was unaffected by the immune challenge, but an advanced moult onset resulted in a longer moult duration. Moreover, female (but not male) throat feather colouration of LPS‐injected individuals showed a reduced UV chroma. We argue that an early activation of the immune system caused by LPS may allow nestlings to better cope with post‐fledging stresses and lead to an earlier moult onset. The effect of early LPS exposure was remarkably persistent, as it was still visible more than one year after the treatment, and highlighted the importance of early developmental stresses in shaping subsequent major life‐history traits, including the timing of moult in birds.     

Bigger brains cycle faster before neurogenesis begins: a comparison of brain development between chickens and bobwhite quail

The chicken brain is more than twice as big as the bobwhite quail brain in adulthood. To determine how this species difference in brain size emerges during development, we examined whether differences in neurogenesis timing or cell cycle rates account for the disparity in brain size between chickens and quail. Specifically, we examined the timing of neural events (e.g. neurogenesis onset) from Nissl-stained sections of chicken and quail embryos. We estimated brain cell cycle rates using cumulative bromodeoxyuridine labelling in chickens and quail at embryonic day (ED) 2 and at ED5. We report that the timing of neural events is highly conserved between chickens and quail, once time is expressed as a percentage of overall incubation period. In absolute time, neurogenesis begins earlier in chickens than in quail. Therefore, neural event timing cannot account for the expansion of the chicken brain relative to the quail brain. Cell cycle rates are also similar between the two species at ED5. However, at ED2, before neurogenesis onset, brain cells cycle faster in chickens than in quail. These data indicate that chickens have a larger brain than bobwhite quail mainly because of species differences in cell cycle rates during early stages of embryonic development.     

Parallel changes in genetic diversity and species diversity following a natural disturbance

We examined the spatial and temporal variation of species diversity and genetic diversity in a metacommunity comprising 16 species of freshwater gastropods. We monitored species abundance at five localities of the Ain river floodplain in southeastern France, over a period of four years. Using 190 AFLP loci, we monitored the genetic diversity of Radix balthica , one of the most abundant gastropod species of the metacommunity, twice during that period. An exceptionally intense drought occurred during the last two years and differentially affected the study sites. This allowed us to test the effect of natural disturbances on changes in both genetic and species diversity. Overall, local (alpha) diversity declined as reflected by lower values of gene diversity H _S and evenness. In parallel, the among-sites (beta) diversity increased at both the genetic ( F _ST) and species ( F _STC) levels. These results suggest that disturbances can lead to similar changes in genetic and community structure through the combined effects of selective and neutral processes.     

Evolutionary aspects of population structure for molecular and quantitative traits in the freshwater snail Radix balthica

Detecting the action of selection in natural populations can be achieved using the QST-FST comparison that relies on the estimation of FST with neutral markers, and QST using quantitative traits potentially under selection. QST higher than FST suggests the action of directional selection and thus potential local adaptation. In this article, we apply the QST-FST comparison to four populations of the hermaphroditic freshwater snail Radix balthica located in a floodplain habitat. In contrast to most studies published so far, we did not detect evidence of directional selection for local optima for any of the traits we measured: QST calculated using three different methods was never higher than FST. A strong inbreeding depression was also detected, indicating that outcrossing is probably predominant over selfing in the studied populations. Our results suggest that in this floodplain habitat, local adaptation of R. balthica populations may be hindered by genetic drift, and possibly altered by uneven gene flow linked to flood frequency.     

Infectivity of Diplostomum spp. in Arctic charr: aspects of exposure duration and cercariae morphology

The diplostomid flukes, Diplostomum spp., infect fish and cause cataract opacities in the eye lens. The effect of exposure dose on abundance of Diplostomum spp. eye flukes in fish is known, but the effect of the duration of cercariae exposure has not been studied. However, under natural conditions, the temporal window for a successful cercaria attachment on fish is very short and, consequently, differences in infectivity of eye fluke cercariae, in the short-exposure durations of a few seconds, are probably biologically the most meaningful. We investigated infectivity of Diplostomum spp. cercariae originating from snail hosts in 3 lakes (3 Lymnaea stagnalis populations and 1 Radix balthica population) in 6 exposure times, ranging from 5 sec to 15 min, in young-of-the-year Arctic charr Salvelinus alpinus. In addition, we compared the infectivity to the cross-morphology of the cercariae. In the long-exposure duration, i.e., > or = 5 min, infectivity of Diplostomum spp. did not vary between the snail host species (L. stagnalis and R. balthica) of the same lake or across the L. stagnalis populations of 3 different lakes. However, in the short-exposure duration, i.e., < or = 60 sec, Diplostomum spp. cercariae shed from L. stagnalis had higher infectivity than did cercariae from R. balthica of the same lake. This indicates that that there is an interaction between length of cercariae exposure and origin of Diplostomum spp., and that the duration of exposure may influence the results when fish are experimentally infected. Within a lake, cercariae shed from L. stagnalis were also smaller than cercariae shed from R. balthica.     

内蒙科尔沁草原淡水螺吸虫幼虫期的调查研究

本文报道在内蒙科尔沁草原检查5种淡水螺:耳萝卜螺、卵萝卜螺,豆螺、凸旋螺及半球多脉扁螺共5,616粒。从中查出至少12种吸虫幼虫期,它们是:土耳其斯坦东毕吸虫、有棘血居吸虫、马蹄吸虫、斜睾吸虫、二种背孔吸虫、棘口类吸虫、重盘吸虫、球孔吸虫、异形科吸虫、杯叶科吸虫及鹗形吸虫。其中东毕吸虫、血居吸虫及棘口吸虫的尾蚴分别进行实验动物的感染试验,获得成虫。各吸虫侵袭期幼虫季节动态亦经观察。     

Flexibility in the timing of post‐breeding moult in passerines in the UK

Higher temperatures resulting from climate change have led to predictions that the duration of the breeding season of many temperate bird species may be changing. However, the extent to which breeding seasons can be altered will also depend on the degree of flexibility in processes occurring at other points in the annual cycle. In particular, plasticity in the timing of post‐breeding moult (PBM) could facilitate changes in the timing of key events throughout the annual cycle, but little is known about the level of within‐ and between‐species plasticity in PBM. As part of the British Trust for Ornithology (BTO) Ringing Scheme, many ringers routinely record moult scores of flight feathers, and these can be used to provide information on the annual progression of PBM for a range of species. Here we use ringing data to investigate patterns of PBM in 15 passerines, as well as data from the BTO Nest Record Scheme to relate these differences to the timing of breeding of these species across the UK. We find considerable variation in both the mean start (19 May–29 July) and duration (66–111 days) of PBM between species, but find no evidence that species starting PBM later in the season complete it any faster. However, there is considerable within‐species variation in PBM, particularly for multi‐brooded species; PBM starts later and is completed in less time when the duration of the breeding season (difference between first and last nests) is longer. This implies that a later end to breeding can be compensated for by faster PBM, and that advances in breeding could lead to earlier and slower PBM. Our findings suggest that adaptation of PBM in response to climate‐mediated changes in the timing and duration of the breeding season is possible. However, the requirement to complete PBM prior to migration or the onset of winter might constrain the extent to which breeding seasons can lengthen, especially for later nesting species.     

Rhogocytes in gastropod larvae: developmental transformation from protonephridial terminal cells

Rhogocytes, terminal cells of protonephridia, and podocytes of metanephridial systems share an architectural feature that creates an apparent sieving device. The sieve serves to ultrafilter body fluid during the excretion and osmoregulation process carried out by nephridial systems, but its function in rhogocytes is unclear. Rhogocytes are molluscan hemocoelic cells that appear to have various functions related to metabolism of metal ions, including synthesis of hemocyanin in some gastropods and metal detoxification in pteriomorph bivalves. A hypothesis that proposed developmental and possibly evolutionary conversion between protonephridial terminal cells and rhogocytes has never been further explored; indeed, information on the occurrence of rhogocytes in molluscan developmental stages is meager. We used transmission electron microscopy to show that rhogocytes are present within larvae of eight species of gastropods sampled from the three major gastropod clades with a feeding larval stage in the life history. In larvae of a heterobranch gastropod, a rhogocyte was located next to each terminal cell of a pair of protonephridia that flanked the foregut, whereas all six species of caenogastropod larvae and a neritimorph larva that we examined had rhogocytes, but no protonephridia, in this location. We did not find ring‐shaped profiles of hemocyanin decamers within rhogocytes of larvae or pre‐hatch embryos. Rhogocytes in newly released larvae of Nerita melanotragus contained orderly bundles of cylinders, but the diameter of the cylinders was only 70% of the diameter typical of hemocyanin multidecamers. By examining embryos of the caenogastropod Nassarius mendicus at four successive developmental time points that bracketed the occurrence of larval hatching, we found that terminal cells from non‐functional protonephridia in pre‐hatch embryos transformed into rhogocytes around the time of hatching. This empirical evidence of ontogenetic transformation of protonephridial terminal cells into rhogocytes might be interpreted as developmental recapitulation of an evolutionary transition that occurred early in molluscan history.     

Early development of johnny darter,Etheostoma nigrum,and fantail darter,E. flabellare,with a discussion of its ecological and evolutionary aspects

Synopsis Fantail darters,Etheostoma flabellare, have larger eggs with greater yolk volume than johnny darters,E. nigrum, and have a longer embryo period. This longer embryo period is associated with accelerated finfold differentiation, producing a larger, better formed individual at the onset of exogenous feeding. Developmental differences among these two species and the rainbow darter,Etheostoma caeruleum, primarily in timing of ontogenetic events, are described in detail. These differences in timing are considered largely early developmental phenomena as they affect embryos, larvae, and aievins but apparently not juveniles or adults. Demographic aspects of early life history were assessed using data from studies of Ontario streams. Differences in early survival among species appear to play an important role in recruitment of mature females, but cannot be attributed to parental care or egg size differences.