Altered thyroid hormone levels affect the capacity for temperature-induced developmental plasticity in larvae of Rana temporaria and Xenopus laevis

Anuran larvae show phenotypic plasticity in age and size at metamorphosis as a response to temperature variation. The capacity for temperature-induced developmental plasticity is determined by the thermal adaptation of a population. Multiple factors such as physiological responses to changing environmental conditions, however, might influence this capacity as well. In anuran larvae, thyroid hormone (TH) levels control growth and developmental rate and changes in TH status are a well-known stress response to sub-optimal environmental conditions. We investigated how chemically altered TH levels affect the capacity to exhibit temperature-induced developmental plasticity in larvae of the African clawed frog (Xenopus laevis) and the common frog (Rana temporaria). In both species, TH level influenced growth and developmental rate and modified the capacity for temperature-induced developmental plasticity. High TH levels reduced thermal sensitivity of metamorphic traits up to 57% (R. temporaria) and 36% (X. laevis). Rates of growth and development were more plastic in response to temperature in X. laevis (+30%) than in R. temporaria (+6%). Plasticity in rates of growth and development is beneficial to larvae in heterogeneous habitats as it allows a more rapid transition into the juvenile stage where rates of mortality are lower. Therefore, environmental stressors that increase endogenous TH levels and reduce temperature-dependent plasticity may increase risks and the vulnerability of anuran larvae. As TH status also influences metabolism, future studies should investigate whether reductions in physiological plasticity also increases the vulnerability of tadpoles to global change.     

Adaptive phenotypic plasticity and genetics of larval life histories in two Rana temporaria populations

Phenotypic plasticity provides means for adapting to environmental unpredictability. In terms of accelerated development in the face of pond-drying risk, phenotypic plasticity has been demonstrated in many amphibian species, but two issues of evolutionary interest remain unexplored. First, the heritable basis of plastic responses is poorly established. Second, it is not known whether interpopulational differences in capacity to respond to pond-drying risk exist, although such differences, when matched with differences in desiccation risk would provide strong evidence for local adaptation. We investigated sources of within- and among-population variation in plastic responses to simulated pond-drying risk (three desiccation treatments) in two Rana temporaria populations originating from contrasting environments: (1) high desiccation risk with weak seasonal time constraint (southern population); and (2) low desiccation risk with severe seasonal time constraint (northern population). The larvae originating from the environment with high desiccation risk responded adaptively to the fast decreasing water treatment by accelerating their development and metamorphosing earlier, but this was not the case in the larvae originating from the environment with low desiccation risk. In both populations, metamorphic size was smaller in the high-desiccation-risk treatment, but the effect was larger in the southern population. Significant additive genetic variation in development rate was found in the northern and was nearly significant in the southern population, but there was no evidence for genetic variation in plasticity for development rates in either of the populations. No genetic variation for plasticity was found either in size at metamorphosis or growth rate. All metamorphic traits were heritable, and additive genetic variances were generally somewhat higher in the southern population, although significantly so in only one trait. Dominance variances were also significant in three of four traits, but the populations did not differ. Maternal effects in metamorphic traits were generally weak in both populations. Within-environment phenotypic correlations between larval period and metamorphic size were positive and genetic correlations negative in both populations. These results suggest that adaptive phenotypic plasticity is not a species-specific fixed trait, but evolution of interpopulational differences in plastic responses are possible, although heritability of plasticity appears to be low. The lack of adaptive response to desiccation risk in northern larvae is consistent with the interpretation that selection imposed by shorter growing season has favored rapid development in north (approximately 8% faster development in north as compared to south) or a minimum metamorphic size at the expense of phenotypic plasticity.     

The correlation between the heat resistance of the zygotes of the common frog Rana temporaria and its embryos at different developmental stages

A study has been made of a correlation between the heat resistance of zygotes and embryos of the same clutches at different stages of development of Rana temporaria L. The embryos were incubated at 19 degrees C, the injurious temperature being 37 degrees C. As criterion of heat resistance served the time of the injurious temperature action which leads by the ++stra of cleavage to a 50 per cent elimination of embryos (LD50). The correlation in question has been evaluated by the rank correlation coefficient (rho). The correlation between the heat resistance of zygotes and embryos at the start of gastrulation (stage 11) was moderate (rho = 0.47). At one of neurulation stages (stage 22) this correlation was weak (rho = 0.207), to become rather high just before the cleavage (stage 28, rho = 0.63). It is assumed that the thermal selection of zygotes, i.e. of the organism at the cellular stage of development, may be oriented to result in the survival of embryos, which just before the cleavage will show both elevated heat resistance and high heritability of this character.     

Quantitative genetic analysis of larval life history traits in two alpine populations of Rana temporaria

We estimated genetic and maternal variance components of larval life history characters in alpine populations of Rana temporaria (the common frog) using a full-sib/half-sib breeding design. We studied trait plasticity by raising tadpoles at 14 or 20°C in the laboratory. Larval period and metamorphic mass were greater at 14°C. Larval period did not differ between populations, but high elevation metamorphs were larger than low elevation metamorphs. Significant additive variation for larval period was detected in the low altitude population. No significant additive variation was detected for mass at metamorphosis (MM), which instead displayed significant maternal effects. Plasticity in metamorphic mass of froglets was greater in the high altitude population. The plastic response of larval period to temperature did not differ between the populations. Evolution of metamorphic mass is likely constrained by lack of additive genetic variation. In contrast, significant heritability for larval period suggests this trait may evolve in response to environmental change. These results differ from other studies on R. temporaria, suggesting that populations of this broadly distributed species present substantial geographic variation in the genetic architecture and plasticity of tadpole life history traits.     

Cross fostering in mice: behavioral and physiological carry-over effects in adulthood

Cross fostering is a widely used laboratory practice. However, relatively few studies have directly investigated the carry-over effects of this procedure in adult animals. The aim of the present study is to investigate the late effects of cross fostering (CF) at birth (in litters composed of no siblings) on adult mice. When adults, cross-fostered male and female mice were examined for intrasex aggression, and levels of emotionality, exploration and anxiety. In addition, body weight was monitored, several internal organs were weighed and plasma corticosterone levels were measured. When compared to controls, body weight of CF male and female mice was increased, at least after early puberty. CF males showed smaller preputial glands, while basal corticosterone level was not affected by cross fostering. In the free-exploratory test, CF males, but not females, showed a behavioral profile suggestive of lower anxiety. These effects in adulthood cannot be ascribed to differences in the maternal care received, which was not affected by cross fostering. In conclusion, cross fostering at birth induced a number of behavioral and physiological alterations in mice, particularly in males. These findings should be carefully evaluated when applying cross fostering procedure to laboratory animals.     

Transgenerational and developmental plasticity at the molecular level: Lessons from Daphnia

Listen to the news and you are bound to hear that researchers are increasingly interested in the biological manifestations of trauma that reverberate through the generations. Research in this area can be controversial in the public realm, provoking societal issues about personal responsibility (are we really born free or are we born with the burden of our ancestors’ experience?). It is also a touchy subject within evolutionary biology because it provokes concerns about Lamarckianism and general scepticism about the importance of extra‐genetic inheritance (Laland et al., 2014 ). Part of why the research in this area has been controversial is because it is difficult to study. For one, there is the problem of how long it takes to track changes across generations, making long‐term, multi‐generational studies especially tricky in long‐lived species. Moreover, there are presently very few (if any) known molecular mechanisms by which environmental effects can be incorporated into the genome and persist for multiple successive generations, casting doubt on their evolutionary repercussions. Fortunately, you only have to look in your local pond to find the creatures that are teaching us a great deal about how and why the experiences of parents are passed down to their offspring. In this issue of Molecular Ecology, Hales et al. (Hales et al., 2017 ) illustrate the power of Daphnia (“water fleas”) for making headway in this field.     

Regulatory effects of homoeologous chromosome arms on wheat proteins at two developmental stages

Summary Two-dimensional gel electrophoresis was conducted on denatured proteins of the 10-day-old first leaf (1F stage) of 18 homoeologous ditelosomic (DT) lines of wheat cultivar Chinese Spring. The observations, compared to the euploid control and relative to previous data found on 7-day-old etiolated seedlings (G7 stage) of the same lines lead to the following statements: 1) the structural genes of 24 spots can be assigned to 12 chromosome arms; 2) regulatory effects are completely different between the 1F and the G7 stages which may indicate that the regulation of protein amounts is often stage-specific; 3) no case of complete gene dosage compensation is observed among 4 groups of hypothesized homoeoallelic products; 4) homoeologous DT lines do not manifest similar effects which suggest the absence of homoeology for the detected regulatory effects.     

Variation in the degree and costs of adaptive phenotypic plasticity among Rana temporaria populations

Adaptive phenotypic plasticity in the form of capacity to accelerate development as a response to pond drying risk is known from many amphibian species. However, very little is known about factors that might constrain the evolution of this type of plasticity, and few studies have explored to what degree plasticity might be constrained by trade-offs dictated by adaptation to different environmental conditions. We compared the ability of southern and northern Scandinavian common frog (Rana temporaria) larvae originating from 10 different populations to accelerate their development in response to simulated pond drying risk and the resulting costs in metamorphic size in a factorial laboratory experiment. We found that (i) northern larvae developed faster than the southern larvae in all treatments, (ii) a capacity to accelerate the response was present in all five southern and all five northern populations tested, but that the magnitude of the response was much larger (and less variable) in the southern than in the northern populations, and that (iii) significant plasticity costs in metamorphic size were present in the southern populations, the plastic genotypes having smaller metamorphic size in the absence of desiccation risk, but no evidence for plasticity costs was found in the northern populations. We suggest that the weaker response to pond drying risk in the northern populations is due to stronger selection on large metamorphic size as compared with southern populations. In other words, seasonal time constraints that have selected the northern larvae to be fast growing and developing, may also constrain their innate ability for adaptive phenotypic plasticity.     

Testing the role of phenotypic plasticity for local adaptation: growth and development in time-constrained Rana temporaria populations

Phenotypic plasticity can be important for local adaptation, because it enables individuals to survive in a novel environment until genetic changes have been accumulated by genetic accommodation. By analysing the relationship between development rate and growth rate, it can be determined whether plasticity in life-history traits is caused by changed physiology or behaviour. We extended this to examine whether plasticity had been aiding local adaptation, by investigating whether the plastic response had been fixed in locally adapted populations. Tadpoles from island populations of Rana temporaria, locally adapted to different pool-drying regimes, were monitored in a common garden. Individual differences in development rate were caused by different foraging efficiency. However, developmental plasticity was physiologically mediated by trading off growth against development rate. Surprisingly, plasticity has not aided local adaptation to time-stressed environments, because local adaptation was not caused by genetic assimilation but on selection on the standing genetic variation in development time.     

The effects of iodine level and source on iodine carry-over in eggs and body tissues of laying hens

In the presented study the effect of different iodine (I) levels and sources in hen feed on the iodine concentration of different tissues, blood serum, and eggs of laying hens was studied. For this purpose, two experiments were conducted with 30 laying hens each. In these experiments feed was enriched with KI and Ca(IO(3))(2), respectively, at 0 (Control), 0.25, 0.5, 2.5 and 5 mg I/kg feed, resulting a analysed iodine level from 0.44 to 4.20 mg/kg feed. After four weeks experimental feeding the iodine concentrations of thyroid glands, blood, meat, liver, abdominal fat and eggs were measured with inductively coupled plasma mass spectrometry. The experimental treatment did not affect hen performance. The iodine supplementation significantly increased the iodine concentration of eggs (144-1304?μg/kg), thyroid glands (3367-5975?μg/g), blood serum (16-67?μg/kg) and liver (13-43?μg/kg). Meat (about 14?μg I/kg) and abdominal fat (about 12?μg?I/kg) were not significantly affected by iodine treatment. Comparative regression analyses showed that at a similar iodine intake, the supply via KI resulted in significantly higher iodine deposition into eggs than Ca(IO(3))(2). Due to the high carry-over of iodine into eggs, eggs may considerably contribute to the iodine supply of the consumers.     

Phenotypic plasticity of body size in Drosophila: effects of a daily periodicity of growth temperature in two sibling species

Abstract. Variation of wing and thorax length under thermoperiodic growth conditions was analysed in four strains of two sibling species, Drosophila melanogaster and D. simulans , from two European localities. Results were compared to those obtained with constant temperatures ranging from 12 to 31 °C.
Under constant temperatures the data basically confirmed previous results: concave reaction norms for wing and thorax length; a monotonically decreasing norm for wing : thorax ratio; and an increasing norm for sex dimorphism (female : male ratio). Phenotypic variability was maximum at extreme temperatures and minimum at middle ones. Slight differences were observed according to the geographical origin: the difference between strains from Bordeaux (France) and Cordoba (Spain) was maximum at low temperatures but disappeared at about 28 °C.
According to the temperatures chosen, alternating thermal regimens had either no effect or produced a significant size reduction, probably reflecting a periodic stress. The magnitude of this effect was proportional to the amplitude of the thermoperiod but not to the quality (cold or heat) of the stress. In a similar way, the wing : thorax ratio was either not modified or reduced significantly, indicating that wing length was relatively more affected than thorax length by alternating thermal regimens. Sex dimorphism also showed either no change or a significant increase, indicating that males were relatively more reactive than females to alternating conditions. Finally, regimens of broad amplitudes increased the phenotypic variability, again an indication of stressful effects. All these observations should be taken into account when analysing phenotypic variability in nature and trying to understand natural selection in wild-living populations.     

Maintenance of brain thyroid hormone level during peripheral hypothyroid condition in adult rat

Thyroid hormones are essential for normal functioning of adult mammalian brain. The present investigation deals with the understanding of the time course of thyroid hormone homeostasis in adult rat brain. Animals were rendered hypothyroid by PTU injections (2 mg/100 g bw) for 30 consecutive days. Serum and synaptosomal T3/T4 content, synaptosomal AChE and Na+-K+-ATPase activities were determined on alternate days. While serum T4 level initially increased on the second day compared to control, serum T3 declined in a triphasic pattern; the first phase lasting from the second day to the 6th day, the second phase ended on the 14th day and last phase continued till the 30th day. Cerebro-cortical synaptosomal T3 level increased on the 2nd day from the control, attained a peak on the 4th day, remained stable until the 18th day, and abruptly declined on the 20th day. Synaptosomal T4 content remained negligible or undetected throughout. Synaptosomal membrane Na+-K+-ATPase and AChE activity exhibited an inverse relationship during the experimental regime, being much more prominent on the 2nd, 18th and 20th day coinciding with the variations in brain T3 level. Thus, the study identifies the onset of central homeostasis between the first and second day, its continuation for about 16-18 days and its termination between the 18th and 20th day.     

Population‐specific effects of developmental temperature on body condition and jumping performance of a widespread European frog

All physiological processes of ectotherms depend on environmental temperature. Thus, adaptation of physiological mechanisms to the thermal environments is important for achieving optimal performance and fitness. The European Common Frog, Rana temporaria, is widely distributed across different thermal habitats. This makes it an exceptional model for studying the adaptations to different thermal conditions. We raised tadpoles from Germany and Croatia at two constant temperature treatments (15°C, 20°C), and under natural temperature fluctuations (in outdoor treatments), and tested how different developmental temperatures affected developmental traits, that is, length of larval development, morphometrics, and body condition, as well as jumping performance of metamorphs. Our results revealed population‐specific differences in developmental time, body condition, and jumping performance. Croatian frogs developed faster in all treatments, were heavier, in better body condition, and had longer hind limbs and better jumping abilities than German metamorphs. The populations further differed in thermal sensitivity of jumping performance. While metamorphs from Croatia increased their jumping performance with higher temperatures, German metamorphs reached their performance maximum at lower temperatures. These population‐specific differences in common environments indicate local genetic adaptation, with southern populations being better adapted to higher temperatures than those from north of the Alps.     

Phenotypic plasticity of body pigmentation in Drosophila: influence of a developmental thermoperiodic regime in two sibling species

Abstract Variation of body pigmentation according to growth temperature was analysed in two European populations (Bordeaux and Cordoba) of the two sibling species Drosophila melanogaster (Meigen) and D. simulans (Sturtevant). Results obtained at constant temperatures were compared with those observed under alternating thermal regimes of various amplitudes and with a daily periodicity of 12–12 h. Abdominal pigmentation in segments 5, 6 and 7, and of their sum, was investigated in both species in females only. Thoracic trident pigmentation was studied in males and females but in Drosophila melanogaster only.
Thoracic pigmentation exhibited curvilinear convex reaction norms under constant conditions, with a darker pigmentation in the population (Bordeaux) living in a colder climate. No specific effects of alternating regimes were seen. For abdomen pigmentation, slightly different reaction norms were observed under constant conditions between Bordeaux and Cordoba strains. There were also slight differences between the two species. Alternating temperatures produced a significant decrease in average pigmentation, roughly proportional to the thermal amplitude.
Alternating stressing conditions induced also an increase in the phenotypic variability of abdominal pigmentation, especially in segments 6 and 7. Finally, because an increase of growth temperature results in a monotonically decreasing reaction norm of both abdomen pigmentation in females and a size-related trait, the wing/thorax ratio, we found a linear relationship between the two variables, with a very high overall correlation at constant temperatures ( r  = 0.99). The correlation was slightly less under alternating conditions, due to a broader dispersal of the experimental values around the regression lines.     

Context-dependent aggregation in Common Frog Rana temporaria tadpoles: influence of developmental stage, predation risk and social environment

1. This study examines the aggregation behaviour and activity of larvae of the Common Frog Rana temporaria in relation to the early social environment, ontogeny and the presence of chemical cues from a predatory fish to address three main questions: (i) Does previous social interaction influence aggregation behaviour in later developmental stages? (ii) To what extent does aggregation behaviour depend upon the risk level perceived by the individual? (iii) Does aggregation behaviour change through ontogeny?
2. Tadpoles were reared from eggs either in small groups or in complete isolation. In relatively early stages of development, tadpoles showed no preference for the side of the test container containing siblings over the side containing no larvae regardless of the social context experienced (isolation or contact with siblings).
3. The presence of chemical cues from a potential predator did not trigger the aggregation behaviour of tadpoles, but it had a remarkable effect on their activity and spatial distribution. Tadpoles exposed to water preconditioned by a predator spent significantly less time swimming and avoided the central area of the test container more frequently than did controls exposed to unconditioned water.
4. Tadpoles were more active, avoided the central section and associated preferentially with conspecifics (siblings) at later stages of development regardless of the social conditioning they had previously experienced.
5. Individuals reared in groups were twice as active as individuals reared in isolation. This suggests that the early social environment experienced by larvae can influence future behaviour and thereby growth and development rates.
6. The expression of conspecific attraction is probably linked to an ontogenetic shift in larval behaviour. However, reduced activity, rather than aggregation, appears to be the basic antipredator mechanism in larval Common Frog.