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PubMed1979

Developmental changes of sepiapterin synthase activity associated with a variegated purple gene in Drosophila melanogaster.

A variegated position effect on the autonomous gene, purple, has been studid enzymologically in Drosophila melanogaster. Sepiapterin synthase, the enzyme system associated with pr+, was examined for activity in different developmental stages of the fly. The results indicate that T(y:22)prc5, cn/prc4 cn flies (flies in which pr+ has been translocated and which exhibit variegation) have a reduced amount of enzyme activity as compared with both Oregon-R and pr1 flies. This reduction in activity was not found in larval stages, which suggests that the inactivation process probably occurs in late larval or early pupal stages. The phenotype of the variegated adult has white eyes with red-colored spots and patches where drosopterins occur. The phenotype of the fly carrying the translocation is modified by the presence of additional Y chromosomes. This extends the observation from other systems that extra heterochromatin acts to suppress the variegated position effect. The advantages of studying the variegation by measuring enzyme activity, as well as the phenotypic expression, are several; for example, the developmental time at which variegation occurs may be estimated even though drosopterin synthesis is not occurring.

Animals↗

PubMed1995

Physiological genetics of the response to a high-sucrose diet by Drosophila melanogaster.

A diet medium containing 10% (w/v) sucrose can be inferred to be stressful to Drosophila melanogaster from the increased developmental time and reduced size and fecundity of emerging flies. The metabolic basis for this stress and the genetic response to it are of interest from the point of view of both metabolic regulation and the evolutionary genetics of adaptation to stress. Here the effects of a high-sucrose diet on live weight, total protein, stored lipid and glycogen, and crude activities of 12 enzymes involved in energy metabolism were quantified. Assays were done on a large population of Drosophila that had been acclimated to the laboratory. A collection of eggs was divided to produce two replicate populations maintained on standard medium and two replicates maintained on high-sucrose medium for 133 generations. At the end of this period, both control and sucrose-selected populations were tested on standard and on high-sucrose medium. Results showed that the immediate effect of the high-sucrose diet (compared to standard medium) for both populations was a reduction in live weight and total protein, and activities of many of the enzymes were also reduced by the sucrose treatment, even after adjusting for the weight effect. Selection resulted in several changes on both the standard and the sucrose medium, but the direction of change was not always the same as the acute effect. In no case was there a significant medium by selection-treatment interaction. The pattern of phenotypic correlations did not resolve the reasons for the direction of the genetic responses. Correlations were generally stable across diets and after selection, but there were notable exceptions.

Adaptation, Physiological↗

PubMed1989

Regulation of pteridine biosynthesis and aromatic amino acid hydroxylation in Drosophila melanogaster.

The relationship between high dietary levels of aromatic amino acid and regulation of pteridines in Drosophila eyes was examined by measuring changes in pool levels of six pterins in the wild type and mutants and amino acid pool levels in flies that carry mutations for pteridine biosynthesis. The effect upon relative viability and developmental times was also analyzed; relative viability was affected by L-phenylalanine, L-tryptophan, and L-tyrosine in decreasing order and the D-amino acids had little or no effect. The changes in concentration of biopterin, dihydrobiopterin, pterin, sepiapterin, drosopterins, and isoxanthopterin showed a characteristic pattern of increased and/or decreased amounts in response to each of the three L-amino acids. Pterin was regularly increased, and isoxanthopterin decreased. L-Tyrosine caused a 2.1-fold increase in dihydrobiopterin, the largest increase found in this study; L-tryptophan also caused dihydrobiopterin to increase but L-phenylalanine did not. Of 18 eye-color mutants examined, 2 were found to contain high levels of phenylalanine and/or tyrosine, Pu2 and Hnr3. These two mutants, along with prc4 cn/prm2b cn, were shown to be very sensitive to dietary L-phenylalanine, indicating that having low levels of certain pteridines makes them susceptible to toxic effects of these amino acids. Therefore, high levels of aromatic amino acids can perturb the balance among pteridine pools, and low levels of some pteridines in mutants are correlated with the inability to withstand the toxic effects of phenylalanine. From the patterns of change in the pteridines we suggest that tetrahydropterin may also be a cofactor for hydroxylation of phenylalanine, along with tetrahydrobiopterin.

Amino Acids↗

PubMed1980

Experimental epidemiology of schistosomiasis. I. The prepatent period and cercarial production of Schistosoma mansoni in Biomphalaria snails at various constant temperatures.

Laboratory experiments have permitted the quantification of the developmental times (prepatent periods) of Schistosoma mansoni in the snail over the whole possible range of constant temperatures. The basis relationship is satisfactorily described by a hyperbola of the formula y = 268/(x-14.2), y being the minimum time from miracidial infection to cercariae shedding (in days) x the mean temperature, and 14.2 the theoretical temperature threshold (in degrees C). Cercariae production takes place within the limits of +16 degrees C and 35 degrees C, the number of cercariae being low and the mortality of snails high at the extreme values. Long-term alternations between two temperature levels resulted in prepatent periods corresponding exactly to the proportional time-temperature products. However, slight accelerations of up to 7% and more could be observed when the prepatency began in a period of high temperature. The number of cercariae each snail shed during a period of one hour exposure per week decreased from about 1,500 at 18 degrees C to about 250 at 16 degrees C. Shedding ceased completely after 1-2 weeks at 15 degrees C.

Animals↗

PubMed1994

An immunocytochemical marker for hamster retinal ganglion cells.

We examined the specificity and developmental time course of the labelling of retinal ganglion cells in Syrian hamsters by a monoclonal antibody AB5. In adult hamsters, AB5 selectively labelled somata in the ganglion cell layer, dendrites in the inner plexiform layer and axons in the nerve fibre layer. When retinal ganglion cells were retrogradely labelled with DiI prior to AB5 immunocytochemistry, all of the retrogradely labelled retinal ganglion cells in the ganglion cell layer were AB5 immunoreactive, indicating that AB5 labels all classes of ganglion cell in that layer. In retinae depleted of retinal ganglion cells by neonatal optic nerve transections, AB5 did not label any somata or processes, indicating that AB5 specifically labels retinal ganglion cells. During development, AB5 labelling first appeared as a weak staining of cell bodies in the ganglion cell layer on postnatal day 12 (P12; PO = first 24 h following birth) and acquired the staining pattern seen in the adult by postnatal day 14. From the onset of AB5 immunoreactivity, AB5-labelled somata of varying sizes were present across the entire retinal surface. Although AB5 labelled retinal ganglion cell axons in the nerve fibre layer of the retina it did not label the optic nerve or retinal ganglion cell axons in the brain at any age examined. AB5 labelling was also found to be compatible with bromodeoxyuridine immunocytochemistry and, therefore, useful for determining the time of generation of hamster retinal ganglion cells.

Animals↗

PubMed2004

Quantitative genetics of functional characters in Drosophila melanogaster populations subjected to laboratory selection.

What are the genetics of phenotypes other than fitness, in outbred populations? To answer this question, the quantitative-genetic basis of divergence was characterized for outbred Drosophila melanogaster populations that had previously undergone selection to enhance characters related to fitness. Line-cross analysis using first-generation and second-generation hybrids from reciprocal crosses was conducted for two types of cross, each replicated fivefold. One type of cross was between representatives of the ancestral population, a set of five populations maintained for several hundred generations on a two-week discrete-generation life cycle and a set of five populations adapted to starvation stress. The other type of cross was between the same set of ancestral-representative populations and another set of five populations selected for accelerated development from egg to egg. Developmental time from egg to eclosion, starvation resistance, dry body weight and fecundity at day 14 from egg were fit to regression models estimating single-locus additive and dominant effects, maternal and paternal effects, and digenic additive and dominance epistatic effects. Additive genetic variation explained most of the differences between populations, with additive maternal and cytoplasmic effects also commonly found. Both within-locus and between-locus dominance effects were inferred in some cases, as well as one instance of additive epistasis. Some of these effects may have been caused by linkage disequilibrium. We conclude with a brief discussion concerning the relationship of the genetics of population differentiation to adaptation.

Animals↗

PubMed1998

X chromosome inactivation and the Xist gene.

Recent years have seen rapid progress towards understanding the molecular mechanisms involved in X chromosome inactivation (X inactivation). This progress has largely revolved around the discovery of the X inactive specific transcript (Xist) gene, which is known now to represent the master switch locus regulating X inactivation. In adult cells Xist is transcribed exclusively from the inactive X chromosome. The transcript has no apparent protein-coding potential and is retained in the nucleus in close association with the domain occupied by the inactive X chromosome. It is thus thought to represent a functional RNA molecule which acts as the primary signal responsible for the propagation of X inactivation. Developmental regulation of Xist correlates with the developmental timing of X inactivation. Recent results have demonstrated that Xist is both necessary and sufficient for X inactivation. Goals for the future are to understand the mechanism of Xist regulation which underlies the establishment of appropriate X inactivation patterns and to determine how Xist RNA participates in the process of propagating inactivation in cis.

Animals↗

PubMed2000

Thermal gains through collective metabolic heat production in social caterpillars of Eriogaster lanestris.

We investigated thermal characteristics of aggregations of social, tent-building caterpillars of the small eggar moth Eriogaster lanestris (Lepidoptera: Lasiocampidae). The highly synchronous behavior of individuals of the colony has important consequences for their thermal ecology. Air temperature in the tent fluctuates according to the caterpillars' activity: air temperature slowly rises about 2.5-3 degrees C above the surroundings when caterpillars aggregate in the tent after feeding and decreases rapidly when the larvae leave the tent. Thermal energy can be stored for a few hours when ambient temperature drops. Experiments show that metabolic heat production sufficiently explains this effect. As even minor additional heat gain may reduce developmental time, aggregating in the tent may thus confer selective advantages under overcast weather or at night, when behavioral thermoregulation through basking is not possible.

Animals↗

PubMed2005

Generation of an oligonucleotide array for analysis of gene expression in Chlamydomonas reinhardtii.

The availability of genome sequences makes it possible to develop microarrays that can be used for profiling gene expression over developmental time, as organisms respond to environmental challenges, and for comparison between wild-type and mutant strains under various conditions. The desired characteristics of microarrays (intense signals, hybridization specificity and extensive coverage of the transcriptome) were not fully met by the previous Chlamydomonas reinhardtii microarray: probes derived from cDNA sequences (approximately 300 bp) were prone to some nonspecific cross-hybridization and coverage of the transcriptome was only approximately 20%. The near completion of the C. reinhardtii nuclear genome sequence and the availability of extensive cDNA information have made it feasible to improve upon these aspects. After developing a protocol for selecting a high-quality unigene set representing all known expressed sequences, oligonucleotides were designed and a microarray with approximately 10,000 unique array elements (approximately 70 bp) covering 87% of the known transcriptome was developed. This microarray will enable researchers to generate a global view of gene expression in C. reinhardtii. Furthermore, the detailed description of the protocol for selecting a unigene set and the design of oligonucleotides may be of interest for laboratories interested in developing microarrays for organisms whose genome sequences are not yet completed (but are nearing completion).

Animals↗

PubMed2004

Beta-adrenoceptor subtype dependence of chronotropy in mouse embryonic stem cell-derived cardiomyocytes.

Cardiomyocytes derived from embryonic stem cells (ESCM) have potential both as an experimental model for investigating cardiac physiology and as a source for tissue repair. For both reasons it is important to characterise the responses of these cells, and one of the key modulators of contraction is the beta-adrenergic system. We therefore undertook a detailed study of the response of the spontaneous beating rate of ESCM to beta-adrenoceptor (betaAR) stimulation. Embryoid bodies (EBs) were generated from murine ES line E14Tg2a by the hanging drop method, followed by plating. Spontaneously beating areas were seen starting from 9-14 days after differentiation: the experiments described here were performed on EBs between developmental day 19 and 48. Beating cell layers were seeded with charcoal to allow tracking of movement by a video-edge detection system. Experiments were performed in physiological medium containing 1 mM Ca2+ at 37 degrees C. Isoprenaline (Iso) increased beating rate with an EC50 value of 52 nM. Iso (0.3 microM) increased basal rate from 67 +/- 7 beats per minute (bpm) to 138 +/- 18 bpm, P < 0.001, n = 22. At earlier developmental time points the response to Iso was not maintained through 5 min exposure; this spontaneous desensitisation only being observed before day 36. A repeat application of Iso after a wash period of 20 min produced reproducible effects on beating rate. Subtype dependence of the betaAR response was determined by comparing an initial response with a second in the presence of selective beta1- or beta2AR antagonists. In the presence of the specific beta1AR-blocker CGP 20712A (300 nM) the increase in rate with Iso was reduced from 207 +/- 42% of basal to 128 +/- 13%, P < 0.01. With the beta2AR-blocker ICI 118,551 (50 nM) there was no significant change in Iso response. Exposure to the muscarinic agonist, carbachol (10 microM), inhibited the increase in frequency mediated by isoprenaline, but had mixed stimulatory and inhibitory effects on basal rate. This study extends the characterisation of ESCM as a preparation for studying receptor pharmacology, and indicates that the beta1AR is the predominant subtype mediating increases in contraction rate in murine ESCM.

Animals↗

PubMed1997

Sodium current expression during postnatal development of rat outer hair cells.

Outer hair cells of the cultured organ of Corti from newborn rats (0-11 days after birth) were studied in the whole-cell patch-clamp configuration. A voltage-activated sodium current was detected in 97% (n = 109) of the cells at 0-9 days after birth. The properties of this current were: (1) its activation and inactivation kinetics were fast and voltage-dependent, (2) the voltage at half-maximum activation was -45.0 mV, (3) its steady-state inactivation was temperature-sensitive (the half-inactivating voltage was -92.6 mV at 23 degrees C and -84.8 mV at 37 degrees C), (4) the reversal potential (80 mV) was close to the sodium equilibrium potential and currents could be abolished by the removal of extracellular sodium, and (5) tetrodotoxin blocked the current with a Kd of 474 nmol/l. Current amplitudes were up to 1.7 nA at room temperature. Mean current amplitudes showed a developmental time course with a maximum at postnatal days 3 and 7 for outer hair cells from the basal and apical part of the cochlea, respectively. In current-clamp mode cells had membrane potentials of -59.7 +/- 11.7 mV (n = 9). When cells were hyperpolarized by constant current injection, depolarizing currents were able to trigger action potentials. At 18 days after birth, sodium currents were greatly reduced and barely detectable. The results show that, unlike adult outer hair cells, immature outer hair cells regularly express voltage-gated sodium channels. However, due to mismatching of the sodium current inactivation range and membrane potential in vitro, a physiological function appears questionable.

Aging↗

PubMed2000

Neurulation in the pig embryo.

Neurulation is based on a multitude of factors and processes generated both inside and outside the neural plate. Although there are models for a general neurulation mechanism, specific sets of factors and processes have been shown to be involved in neurulation depending on developmental time and rostro-caudal location at which neurulation occurred in the species under investigation. To find a common thread amongst these apparently divergent modes of neurulation another representative mammalian species, the pig, was studied here by scanning electron microscopy. The data are compared to a series of descriptions in other species. Furthermore, the relation of axial curvature and neural tube closure rate is investigated. In the pig embryo of 7 somites, the first apposition of the neural folds occurs at somite levels 5-7. This corresponds to closure site I in the mouse embryo. At the next stage the rostral and caudal parts of the rhombencephalic folds appose, leaving an opening in between. Therefore, at this stage four neuropores can be distinguished, of which the anterior and posterior ones will remain open longest. The two rhombencephalic closure sites have no counterpart in the mouse, but do have some resemblance to those of the rabbit. The anterior neuropore closes in three phases: (1) the dorsal folds slowly align and then close instantaneously, the slow progression being likely due to a counteracting effect of the mesencephalic flexure; (2) the dorso-lateral folds close in a zipper-like fashion in caudo-rostral direction; (3) the final round aperture is likely to close by circumferential growth. At the stage of 22 somites the anterior neuropore is completely closed. In contrast to the two de novo closure sites for the anterior neuropore in the mouse embryo, none of these were detected in the pig embryo. The posterior neuropore closes initially very fast in the somitic region, but this process almost stops thereafter. We suggest that the somites force the neural folds to elevate precociously. Between the stages of 8-20 somites the width of the posterior neuropore does not change, while the rate of closure gradually increases; this increase may be due to a catch-up of intrinsic neurulation processes and to the reduction of axial curvature. At the stage of 20-22 somites the posterior neuropore suddenly reduces in size but thereafter a small neuropore remains for 5 somite stages. The closure of the posterior neuropore is completed at the stage of 28 somites.

Animals↗

PubMed2026

Genetic-epigenetic interactions (meQTLs) in orofacial clefts etiology.

Understanding how genetic variants influence disease risk through molecular mechanisms remains a central challenge in complex disease genetics. Nonsyndromic orofacial clefts (OFCs) exemplify this challenge, with most risk loci residing in non-coding regions. We hypothesized that common genetic variants influence OFC risk by modulating DNA methylation at regulatory elements through methylation quantitative trait loci (meQTLs). We analyzed 10 OFC-associated SNPs against genome-wide DNA methylation profiles in 409 cases and 456 controls, identifying 23 potential meQTLs. Findings were validated using 358 cleft-discordant sibling pairs with MethyLight assays. We performed formal mediation analysis, genotype-tissue interaction and cross-referenced with the mQTL Database to assess developmental timing. Nine meQTLs were validated, including rs987525 (8q24)-cg16561172 (MYC) (P = 9.6 × 10⁻⁶), which mapped to a mesendoderm-active enhancer upstream of MYC. Genotype × tissue interaction confirmed tissue-specificity (P = 1.00 × 10- 3), with stronger effects in oral-derived tissue (saliva). Additional validated SNP-CpG associations involved MAFB-PLCG1, NOG-PPM1E, FOXE1-FRZB, and SPRY2-LGR4. While effect sizes correlated between tissues (r = 0.81), formal mediation analysis indicated individual CpG sites do not fully mediate SNP-phenotype relationships, suggesting coordinated epigenetic mechanisms. Most associations showed peak effects during childhood, while 8q24 showed unique adult-specific patterns. We identified genetic variants influencing methylation at craniofacial regulatory elements, and provided a mechanistic link for a major risk locus, 8q24, with tissue-specific effects in saliva. While individual CpG sites did not fully mediate the genetic risk, our findings identified specific regulatory regions where coordinated epigenetic changes may contribute to OFC susceptibility.

Humans↗

PubMed2005

Development of A-type allatostatin immunoreactivity in antennal lobe neurons of the sphinx moth Manduca sexta.

The antennal lobe (AL) of the sphinx moth Manduca sexta is a well-established model system for studying mechanisms of neuronal development. To understand whether neuropeptides are suited to playing a role during AL development, we have studied the cellular localization and temporal expression pattern of neuropeptides of the A-type allatostatin family. Based on morphology and developmental appearance, we distinguished four types of AST-A-immunoreactive cell types. The majority of the cells were local interneurons of the AL (type Ia) which acquired AST-A immunostaining in a complex pattern consisting of three rising (RI-RIII) and two declining phases (DI, DII). Type Ib neurons consisted of two local neurons with large cell bodies not appearing before 7/8 days after pupal ecdysis (P7/P8). Types II and III neurons accounted for single centrifugal neurons, with type II neurons present in the larva and disappearing in the early pupa. The type III neuron did not appear before P7/P8. RI and RII coincided with the rises of the ecdysteroid hemolymph titer. Artificially shifting the pupal 20-hydroxyecdysone (20E) peak to an earlier developmental time point resulted in the precocious appearance of AST-A immunostaining in types Ia, Ib, and III neurons. This result supports the hypothesis that the pupal rise in 20E plays a role in AST-A expression during AL development. Because of their early appearance in newly forming glomeruli, AST-A-immunoreactive fibers could be involved in glomerulus formation. Diffuse AST-A labeling during early AL development is discussed as a possible signal providing information for ingrowing olfactory receptor neurons.

Animals↗

PubMed2005

Testing models of parental investment strategy and offspring size in ants.

Parental investment strategies can be fixed or flexible. A fixed strategy predicts making all offspring a single 'optimal' size. Dynamic models predict flexible strategies with more than one optimal size of offspring. Patterns in the distribution of offspring sizes may thus reveal the investment strategy. Static strategies should produce normal distributions. Dynamic strategies should often result in non-normal distributions. Furthermore, variance in morphological traits should be positively correlated with the length of developmental time the traits are exposed to environmental influences. Finally, the type of deviation from normality (i.e., skewed left or right, or platykurtic) should be correlated with the average offspring size. To test the latter prediction, we used simulations to detect significant departures from normality and categorize distribution types. Data from three species of ants strongly support the predicted patterns for dynamic parental investment. Offspring size distributions are often significantly non-normal. Traits fixed earlier in development, such as head width, are less variable than final body weight. The type of distribution observed correlates with mean female dry weight. The overall support for a dynamic parental investment model has implications for life history theory. Predicted conflicts over parental effort, sex investment ratios, and reproductive skew in cooperative breeders follow from assumptions of static parental investment strategies and omnipresent resource limitations. By contrast, with flexible investment strategies such conflicts can be either absent or maladaptive.

Animals↗

PubMed2005

Correlation between olfactory responses, dispersal tendencies, and life-history traits of the predatory mite Neoseiulus womersleyi (Acari: Phytoseiidae) of eight local populations.

To investigate the relationship between foraging behavior and life-history traits of the predatory mite Neoseiulus womersleyi, the olfactory responses, dispersal ratios from a prey patch, predation rates, fecundity, and developmental times in eight local populations of N. womersleyi were investigated. Significant differences among local populations were found in all these traits except fecundity. None of the life-history traits correlated with foraging behavior. A significant positive correlation was found only between the olfactory response and the dispersal ratio. These results suggested that predatory mites with low olfactory responses would stay in a prey patch longer than predatory mites with high olfactory responses.

Animals↗

PubMed2005

Influence of selected fruit tree pollen on life history of Euseius stipulatus (Acari: Phytoseiidae).

Euseius stipulatus (Athias-Henriot) is a predatory mite widespread in the Mediterranean region considered to be important for the biological control of spider mites in citrus orchards. Development, survival and reproduction of this phytoseiid mite feeding on seven commercially obtained pollen were studied under constant laboratory conditions (20 +/- 1 degrees C, RH 65 +/- 5%, photoperiod 16L: 8D h). Mites were kept individually at rearing units with ample quantity of almond (Prunus amygdalus Batch), apple (Malus domestica Borkh), apricot (Prunus armeniaca L.), cherry (Prunus avium L.), pear (Pyrus communis L.), plum (Prunus domestica L.) and walnut (Juglans regia L.) pollen as food source. Developmental time from egg to adult varied between the several pollen tested from 8.38 +/-0.08 to 9.58 +/- 0.11 days for females and from 8.23 +/-0.12 and 9.07 +/-0.12 days for males. Female longevity varied from 11.53 +/- 1.22 to 51.38 +/- 2.45 days, while fecundity ranged from 22.84 +/- 2.30 to 43.61 +/- 3.78 eggs/female. The predator was unable to reproduce when feeding on walnut pollen. Data were submitted to life table analysis and values of the intrinsic rate of increase were derived, ranging from 0.079 to 0.146 (day(-1)). The cumulative Weibull function that was used to describe the age specific survival of females produced excellent fits to the survival data. Results show that almond, plum, cherry and apricot pollen possess higher nutritional value for E. stipulatus than pear and apple pollen and thus may contribute in sustaining and increasing the predator population in field conditions. Walnut pollen can be utilized by the predator only to survive during short periods of time when principal or alternative food sources are scarce.

Acari↗

PubMed2005

Sublethal effects of tebufenpyrad on the eggs and immatures of two-spotted spider mite, Tetranychus urticae.

Sublethal effects of the mitochondrial electron transport inhibitor (METI) tebufenpyrad on Tetranychus urticae Koch females surviving treatment as eggs or immatures (> or =90% mortality) were investigated in life-table assay. The developmental time of females that had survived treatment as eggs (2 mg/l) or larvae (2.5 mg/l) was 1 day longer, while that of protonymphs (2.5 mg/l) or deutonymphs (4 mg/l) was 2 days longer, in addition to reduced longevity and fertility, compared to control. The treatment significantly reduced the intrinsic rate of increase in female survivors: corresponding values were 0.258, 0.278, 0.207 and 0.209, respectively (0.307 in untreated females). The offspring age distribution was significantly affected by the treatment. Sublethal effect of tebufenpyrad and its impact on T. urticae management are discussed.

Animals↗