ABSTRACT
Potential influences of the whole benthic organisms' activity (i.e., coupled faunal and microbial effects) on (58)Co, (51)Cr and (65)Zn diffusion into surface mangrove sediment layers (0-6 cm depth) were evaluated in 36 h experiments. Benthic activity indices (BAI) were proposed, calculated as the relative percent difference between untreated sediments and formaldehyde-treated sediments data in relation to untreated sediments data. Benthic activity was estimated as responsible for 32%-44% of total inventories within sediments, being the chromate anion spiked the less affected radiotracer, while (65)Zn was the most sensitive. Benthic activity was quantitatively evidenced as a control on trace metal diffusion into the sediments, contributing to determine the sediment role as a metal sink. This influence can also affect metal potential bioavailability, considering that recently diffused metals can be more readily available to biological uptake.
Subject(s)
Environmental Monitoring , Geologic Sediments/chemistry , Metals/analysis , Radioactive Tracers , Water Pollutants, Chemical/analysis , Wetlands , Animals , DiffusionABSTRACT
RAD52 is required for almost all recombination events in Saccharomyces cerevisiae. We took advantage of the heterozygosity of HIS4 in the Candida albicans SC5314 lineage to study the role of Rad52 in the genomic stability of this important fungal pathogen. The rate of loss of heterozygosity (LOH) at HIS4 in rad52-ΔΔ strains was â¼10(-3) , at least 100-fold higher than in Rad52(+) strains. LOH of whole chromosome 4 or truncation of the homologue that carries the functional HIS4 allele was detected in all 80 rad52-ΔΔ His auxotrophs (GLH -GL lab His(-)) obtained from six independent experiments. Isolates that had undergone whole chromosome LOH, presumably due to loss of chromosome, carried two copies of the remaining homologue. Isolates with truncations carried centric fragments of broken chromosomes healed by de novo telomere addition. GLH strains exhibited variable degrees of LOH across the genome, including two strains that became homozygous for all the heterozygous markers tested. In addition, GLH strains exhibited increased chromosomal instability (CIN), which was abolished by reintroduction of RAD52. CIN of GLH isolates is reminiscent of genomic alterations leading to cancer in human cells, and support the mutator hypothesis in which a mutator mutation or CIN phenotype facilitate more mutations/aneuploidies.
Subject(s)
Candida albicans/genetics , Candida albicans/metabolism , Chromosome Deletion , Chromosomes, Bacterial/genetics , Fungal Proteins/metabolism , Rad52 DNA Repair and Recombination Protein/metabolism , Chromosomes, Bacterial/metabolism , Fungal Proteins/genetics , Genomic Instability , Loss of Heterozygosity , Rad52 DNA Repair and Recombination Protein/geneticsABSTRACT
The size and shape of organs depend on cellular processes such as cell proliferation, cell survival, and spatial arrangement of cells. In turn, all of these processes are a consequence of positional identity of individual cells in whole organs. Links of positional information with organ growth and pattern expression of genes is a little-addressed question. We show that differences in vestigial expression between neighboring cells of the wing blade autonomously and nonautonomously affect cell proliferation along the proximo-distal axis. On the other hand, uniform expression of vestigial inhibits cell proliferation and also perturbs the shape of wing blade altering the preferential orientation of cell divisions. Our observations provide evidence that local cell interactions, triggered by differences in vestigial expression between neighboring cells, confer positional values operating in the control of growth and shape of the wing.
Subject(s)
Body Patterning/genetics , Drosophila Proteins/metabolism , Drosophila melanogaster/growth & development , Nuclear Proteins/metabolism , Wings, Animal/growth & development , Animals , Cell Cycle , Cell Proliferation , Drosophila Proteins/analysis , Drosophila Proteins/genetics , Drosophila melanogaster/chemistry , Drosophila melanogaster/metabolism , Nuclear Proteins/analysis , Nuclear Proteins/genetics , Organ Size , Proto-Oncogene Proteins/metabolism , Signal Transduction , Wings, Animal/chemistry , Wings, Animal/metabolism , Wnt1 ProteinABSTRACT
Anatomical and molecular characters used to differentiate populations of the land snail Cornu aspersum (Helix aspersa) exhibit, in the western Mediterranean, definite and concordant patterns of correlation with geography. Scenarios involving Pliocene geological changes and postglacial expansion during the Pleistocene were proposed in previous studies to account for the establishment of this geographical structure. In the present work, we have performed a spatial analysis of variation in shell morphometrics, after the partitioning of the overall variation into size and shape components by means of a principal component-based approach (Cadima and Jolliffe, 1996). In order to know if the same historical events have also structured shell variation, the analysis includes all the populations from North Africa which were investigated for anatomical and molecular surveys. Contrary to shell size, which shows a significant spatial heterogeneity essentially related to environmental pressures, variation in shell shape components splits the populations according to a geographical pattern reflective of hypotheses suggested for molecular markers and genital anatomy. This implies that the selective forces often invoked to explain spatial changes in shell shape are not the deciding factors in the present case. Moreover, within each of the two geographical clusters defined, Mantel correlograms show that the similarity between populations declines according to an isolation by distance model. Because of the different allometric relationships between shell size and genitalia measurements in Western and Eastern entities of North Africa, mechanical constraints, possibly leading to a precopulatory isolation in the contact zone, are involved.
Subject(s)
Genetic Variation , Helix, Snails/anatomy & histology , Phylogeny , Africa, Northern , Animals , Biological Evolution , DNA, Mitochondrial/analysis , Genetics, Population , Geography , Helix, Snails/geneticsABSTRACT
A conspicuous shell polychromatism is observed in colonies of Cepaea nemoralis from western France (Brittany). The present study is intended to search for a spatial structure of shell features at this scale and to infer evolutionary processes from the observed patterns. We used a database of morph frequencies (six composite phenotypes were retained) measured on 213 samples regularly distributed on the whole studied area. Data analysis was based on two distinct multivariate methods leading to the following steps: (i) to search for a structure without reference to environmental conditions with a method (global principal components analysis: GPCA) which takes into account the spatial information by means of a neighbouring relationship between sampling points (Delaunay triangulation); (ii) to test the structuring power of environmental conditions by means of two explanatory factors (distance from the sea, altitude) involved in a redundancy analysis (RDA); (iii) to search for a spatial structure using residuals of the previous analysis, ie, after removing effects of environmental conditions. Global covariance accounted for 26.4% of the total variance, leading to a highly significant autocorrelation for each phenotype (step 1). Geographical mapping of factorial scores resulting from global analysis showed a well structured littoral zone and a strong southern-northern inland differentiation. Sixteen percent of the total variance was expressed in RDA but all morphs were not equally concerned. After removing environmental effects, a significant spatial structure still remains but was essentially caused by random processes. We argue for the importance of these last phenomena.
Subject(s)
Snails/genetics , Animals , Biological Evolution , France , Genetic Variation , Models, Biological , Models, Statistical , Polymorphism, GeneticABSTRACT
Heritable mutations in the germ line lead to genetically heterogeneous, or mosaic, gonads. Many of the genes used in germ-line development also play roles in somatic development [Saffman, E. E. & Lasko, P. (1999) Cell. Mol. Life Sci. 55, 1141-1163]. Mutations in these genes may have cellular phenotypes throughout germ-line development leading to their differential elimination or survival, as has been observed in somatic cells [Morata, G. & Ripoll, P. (1975) Dev. Biol. 42, 211-221]. We investigate whether mutations in heterozygosis are subject to pregametic selection in the germ line. We initiated clones of wild-type homozygous cells at different stages of development in gonads heterozygous for eight different recessive chromosome deficiencies. Here we show that cell selection takes place in mosaic germ-line populations. This phenomenon represents a level of selection that precedes and conditions subsequent zygotic selection by affecting the genes available in the gametic population.
Subject(s)
Drosophila melanogaster/embryology , Drosophila melanogaster/genetics , Mosaicism/genetics , Spermatozoa/physiology , Zygote/physiology , Animals , Female , Heterozygote , Larva/radiation effects , Male , Mitosis , Ovary/cytology , Ovary/physiology , Recombination, Genetic/radiation effects , Spermatozoa/radiation effects , X-RaysABSTRACT
The genetic structure of the land snail Helix aspersa was investigated for 21 populations collected along a road located in the polders of the Bay of Mont-Saint-Michel (Brittany, France), following a sampling scheme the area of which did not exceed 900 m in length. A total of 369 individuals were genotyped for five enzymatic markers and seven microsatellite loci. We used sequential hierarchical F-statistics at different spatial scales and spatial autocorrelation statistics to explore recent historical patterns involved in the observed genetic distribution. Whatever the statistics used, congruent levels of spatial genetic substructuring across loci were demonstrated, excepted for one allozyme locus. Overall spatial genetic arrangement matched in a substantial fashion theoretical predictions based on the limited dispersal power of land snails. Positive autocorrelation over short-distance classes may result from the development of genetically distinct patches of individuals organized in family-structured colonies. Therefore, spatial signatures of average I correlograms can be viewed as the expression of a stepping-stone model of population structure, sometimes involving external migrational events. Overall, the revealed pattern of population subdivision on a microgeographical scale was suggestive of a neighbourhood structure. Finally, microsatellite loci are especially suitable for the detection of small genetic clustering, and combining different classes of markers offers the potential to gain further insight into the description of spatial genetic variability over short temporal and geographical scales.
Subject(s)
Enzymes/genetics , Genetics, Population , Microsatellite Repeats , Polymorphism, Genetic , Snails/genetics , Animals , Models, GeneticABSTRACT
A survey of the temporal pattern of population structure and feeding activity of the seabird tick Ixodes uriae was conducted for the first time in two host species colonies: King penguin (Aptenodytes patagonicus halli) and Macaroni penguin (Eudyptes chrysolophus chrysolophus). The life cycle of the tick was investigated over 3 years in a King penguin colony and 2 years in a Macaroni penguin colony at Possession Island (Crozet Archipelago). There was a marked seasonal feeding activity pattern of ticks in both host species, connected with the presence of birds during the breeding season. Although the King penguin colonies were occupied throughout the year by birds, the favourable period for engorgement was limited to 3.5-4.5 months, and almost all the ticks overwintered in the unengorged state. Consequently, I. uriae probably completed its life cycle over 3 years in King penguin colonies. In contrast, this life cycle could be shortened to 2 years in Macaroni penguin colonies, as a result of a different timetable of the presence of birds for breeding and moulting. The relationships between such plasticity and the host behaviour and subantarctic climatic conditions are discussed.
Subject(s)
Birds/parasitology , Ixodes/growth & development , Sexual Behavior, Animal , Tick Infestations/parasitology , Animals , Antarctic Regions , Birds/physiology , Female , Geography , Host-Parasite Interactions , Male , Population Dynamics , SeasonsABSTRACT
One of the fundamental events in metamorphosis in insects is the replacement of larval tissues by imaginal tissues. Shortly after pupariation the imaginal discs evaginate to assume their positions at the surface of the prepupal animal. This is a very precise process that is only beginning to be understood. In Drosophila, during embryonic dorsal closure, the epithelial cells push the amnioserosa cells, which contract and eventually invaginate in the body cavity. In contrast, we find that during pupariation the imaginal cells crawl over the passive larval tissue following a very accurate temporal and spatial pattern. Spreading is driven by filopodia and actin bridges that, protruding from the leading edge, mediate the stretching of the imaginal epithelia. Although interfering with JNK (Jun N-terminal kinase) and dpp (decapentaplegic) produces similar phenotypic effects suppressing closure, their effects at the cellular level are different. The loss of JNK activity alters the adhesion properties of larval cells and leads to the detachment of the imaginal and larval tissues. The absence of dpp signaling affects the actin cytoskeleton, blocks the emission of filopodia, and promotes the collapse of the leading edge of the imaginal tissues. Interestingly, these effects are very similar to those observed after interfering with JNK and dpp signaling during embryonic dorsal closure.
Subject(s)
Cell Adhesion/physiology , Cytoskeleton/physiology , Drosophila Proteins , Insect Proteins/metabolism , Mitogen-Activated Protein Kinases/metabolism , Thorax/embryology , Animals , Cell Communication , Drosophila/embryology , Epidermis/embryology , Epithelial Cells , JNK Mitogen-Activated Protein Kinases , Larva , Models, Biological , Pupa , Wings, Animal/embryologyABSTRACT
We have studied several cell behaviour parameters of mutant alleles of fat (ft) in Drosophila imaginal wing disc development. Mutant imaginal discs continue growing in larvae delayed in pupariation and can reach sizes of several times those of wild-type. Their growth is, however, basically allometric. Homozygous ft cells grow faster than their twin cells in clones and generate larger territories, albeit delimited by normal clonal restrictions. Moreover, ft cells in clones tend to grow towards wing proximal regions. These behaviours can be related with failures in cell adhesiveness and cell recognition. Double mutant combinations with alleles of other genes, e.g. of the Epidermal growth factor receptor (DER) pathway, modify ft clonal phenotypes, indicating that adhesiveness is modulated by intercellular signalling. Mutant ft cells show, in addition, smaller cell sizes during proliferation and abnormal cuticular differentiation, which reflect cell membrane and cytoskeleton anomalies, which are not modulated by the DER pathway.
Subject(s)
Drosophila/genetics , Membrane Proteins/genetics , Wings, Animal/growth & development , Wings, Animal/pathology , Alleles , Animals , Cadherins/genetics , Cadherins/metabolism , Cell Differentiation/genetics , Cell Division/genetics , Drosophila/growth & development , ErbB Receptors/metabolism , Gene Expression Regulation, Developmental , Hyperplasia , Larva , Membrane Proteins/metabolism , Mosaicism , Mutation , Regeneration , Signal TransductionABSTRACT
The function of extramacrochaetae is required during the development of the Drosophila wing in processes such as cell proliferation and vein differentiation. extramacrochaetae encodes a transcription factor of the HLH family, but unlike other members of this family, Extramacrochaetae lacks the basic region that is involved in interaction with DNA. Some phenotypes caused by extramacrochaetae in the wing are similar to those observed when Notch signalling is compromised. Furthermore, maximal levels of extramacrochaetae expression in the wing disc are restricted to places where Notch activity is higher, suggesting that extramacrochaetae could mediate some aspects of Notch signalling during wing development. We have studied the relationships between extramacrochaetae and Notch in wing development, with emphasis on the processes of vein formation and cell proliferation. We observe strong genetic interaction between extramacrochaetae and different components of the Notch signalling pathway, suggesting a functional relationship between them. We show that the higher level of extramacrochaetae expression coincides with the domain of expression of Notch and its downstream gene Enhancer of split-m(beta). The expression of extramacrochaetae at the dorso/ventral boundary and in boundary cells between veins and interveins depends on Notch activity. We propose that at least during vein differentiation and wing margin formation, extramacrochaetae is regulated by Notch and collaborates with other Notch-downstream genes such as Enhancer of split-m(beta).
Subject(s)
DNA-Binding Proteins/genetics , Drosophila Proteins , Membrane Proteins/genetics , Repressor Proteins , Signal Transduction , Wings, Animal/growth & development , Animals , Basic Helix-Loop-Helix Transcription Factors , DNA-Binding Proteins/metabolism , Drosophila/genetics , Drosophila/growth & development , Insect Proteins/genetics , Insect Proteins/metabolism , Receptors, NotchABSTRACT
Notch signaling is involved in cell differentiation and patterning during morphogenesis. In the Drosophila wing, Notch activity regulates the expression of several genes at the dorsal/ventral boundary, and this is thought to elicit wing-cell proliferation. In this work, we show the effect of clones of cells expressing different forms of several members of the Notch signaling pathway, which result in an alteration of Notch activity. The ectopic expression in clones of activated forms of Notch or of its ligands (Delta or Serrate) in the wing causes outgrowths associated with the appearance of ectopic wing margins. These outgrowths consist of mutant territories and of surrounding wild-type cells. However, the ectopic expression of Delta, at low levels in ventral clones, causes large outgrowths that are associated neither with the generation of wing margin structures nor with the expression of genes characteristic of the dorsal/ventral boundary. These results suggest that Notch activity is directly involved in cell proliferation, independently of its role in the formation of the dorsal/ventral boundary. We propose that the nonautonomous effects (induction of extraproliferation and vein differentiation in the surrounding wild-type cells) result from pattern accommodation to positional values caused by the ectopic expression of Notch.
Subject(s)
Drosophila/growth & development , Gene Expression Regulation, Developmental , Membrane Proteins/physiology , Wings, Animal/growth & development , Alleles , Animals , Cell Division/physiology , Drosophila/embryology , Drosophila/genetics , Drosophila Proteins , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mitosis , Phenotype , Promoter Regions, Genetic , Receptors, Notch , Wings, Animal/cytology , Wings, Animal/embryologyABSTRACT
The microspatial genetic structure of allele frequencies at seven isozyme loci was examined for 15 populations of the land snail Helix aspersa sampled in a village from Brittany (north-western France). Spatial heterogeneity of allele frequencies was highly significant (P < 0.001). Fixation indices reflected nonrandom mating within neighbourhoods and a slight but consistent differentiation between colonies (FST=0.044; P < 0.01). Analyses of gene flow or genetic distances failed to reveal a significant relationship with geographical distance, probably because of the complexity of environmental heterogeneity. However, matrix comparisons between genetic distances and connectivity networks among adjacent colonies (Gabriel-connected graph) yielded a significant correlation in every case, indicating a 'step-by-step' relationship between neighbouring localities. Moreover, most of the allozymes were spatially structured and showed (i) a gradual isolation of colonies with increasing geographical distances, and (ii), for some correlogram profiles, a circular gradient illustrating a multidirectional colonization of the village. The probable existence of disperser individuals allowed us to suggest a metapopulation model which would explain the maintenance of such animals in fragmented habitats where anthropogenic disturbances and extinction/recolonization events are commonly observed.
ABSTRACT
The Extramacrochaetae (emc) gene encodes a transcription factor with an HLH domain without the basic region involved in interaction with DNA present in other proteins that have this domain. EMC forms heterodimers with bHLH proteins preventing their binding to DNA, acting as a negative regulator. The function of emc is required in many developmental processes during the development of Drosophila, including wing morphogenesis. Mitotic recombination clones of both null and gain-of-function alleles of emc, indicate that during wing morphogenesis, emc participates in cell proliferation within the intervein regions (vein patterning), as well as in vein differentiation. The study of relationships between emc and different genes involved in wing development reveal strong genetic interactions with genes of the Ras signalling pathway (torpedo, vein, veinlet and Gap), blistered, plexus and net, in both adult wing phenotypes and cell behaviour in genetic mosaics. These interactions are also analyzed as variations of emc expression patterns in mutant backgrounds for these genes. In addition, cell proliferation behaviour of emc mutant cells varies depending on the mutant background. The results show that genes of the Ras signalling pathway are co-operatively involved in the activity of emc during cell proliferation, and later antagonistically during cell differentiation, repressing EMC expression.
Subject(s)
DNA-Binding Proteins/physiology , Drosophila Proteins , Drosophila melanogaster/growth & development , Gene Expression Regulation, Developmental , Insect Proteins/physiology , Repressor Proteins , Transcription Factors/physiology , Wings, Animal/growth & development , Alleles , Animals , Basic Helix-Loop-Helix Transcription Factors , Cell Division , DNA-Binding Proteins/genetics , Drosophila melanogaster/genetics , Genes, ras , Helix-Loop-Helix Motifs , In Situ Hybridization , Insect Proteins/genetics , Morphogenesis/genetics , Mosaicism , Signal Transduction , Transcription Factors/genetics , Veins , Wings, Animal/blood supply , ras Proteins/physiologyABSTRACT
Dorso-ventral axis formation in the Drosophila wing requires the localized accumulation of the Apterous LIM/homeodomain protein (Ap) in dorsal cells. Here we report that dLdb/Chip encodes a LIM-binding cofactor that controls Ap activity. Both lack and excess of dLdb/Chip function cause the same phenotype as apterous (ap) lack of function; i.e. dorsal to ventral transformations, generation of new wing margins, and wing outgrowths. These results indicate that the normal function of Ap in dorso-ventral compartmentalization requires the correct amount of the DLDB/CHIP co-factor, and suggest that the Ap and DLDB/CHIP proteins form a multimeric functional complex. In support of this model, we show that the dLdb/Chip excess-of-function phenotypes can be rescued by ap overexpression.
Subject(s)
Drosophila Proteins , Homeodomain Proteins/metabolism , Insect Proteins/physiology , Nuclear Proteins/physiology , Transcription Factors/metabolism , Animals , Animals, Genetically Modified , Body Patterning , Drosophila/growth & development , Gene Expression , Guinea Pigs , Insect Proteins/genetics , Insect Proteins/metabolism , LIM-Homeodomain Proteins , Mice , Mosaicism , Mutagenesis , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Phenotype , Wings, AnimalABSTRACT
High-resolution two-dimensional (2D) gel electrophoresis coupled with computer analysis has been used to construct a quantitative protein database of Drosophila mature wing imaginal discs. The level of expression for all of the detected proteins has been quantitatively determined. This database has been used to evaluate changes in the patterns of protein synthesis in wing imaginal discs from two Drosophila melanogaster mutants with abnormal wing disc development: fat (ft) and two different alleles of lethal (2) giant disc (l(2)gd). Patterns of pulse-labeled proteins of the different mutants show variations in both qualitative and quantitative parameters of synthesis. In this comparison we have detected specific sets of protein changes characteristic of both alleles of the same locus and a set of protein changes common to both loci. How the abnormal expression of these proteins relates to the abnormal process of mutant hyperplasia is discussed.
Subject(s)
Cadherins , Drosophila Proteins , Drosophila/genetics , Insect Proteins/genetics , Membrane Proteins/genetics , Tumor Suppressor Proteins , Animals , Databases as Topic , Drosophila/metabolism , Electrophoresis, Gel, Two-Dimensional , Genes, Tumor Suppressor/genetics , Insect Proteins/metabolism , Membrane Proteins/metabolism , MutationSubject(s)
Developmental Biology , Drosophila/genetics , Animals , Genes, Insect , Molecular BiologyABSTRACT
The Entelechia model is a generative model of morphogenesis where individual cells exhibit surface labels that express scalar difference and planar polarity along two orthogonal axes X and Y. The amount of surface label depends on the level of Martial (M) gene product within each cell. The model assumes that the confrontation of cells on both sides of compartment borders causes an increase in their level of M gene expression. The resulting disparity between the M value of border cells and that of their neighbors induces the latter to divide. After each division the daughter cells increase their own M value, and allocate to the best matching value position. The increase in M value at the borders therefore extends through the anlage in a cascade of proliferation. The Entelechia condition is reached when the border cells attain the species-specific maximal M values, and the value differences between adjacent cells become indistinguishable. Computer simulations reveal that this model accounts for a variety of observations made on imaginal discs, e.g., 1) each disc attains a constant size in terms of number of cells, independently of the growing conditions; 2) clonal restrictions separate populations of cells which proliferate by intercalar growth; 3) dissociated cells are capable of reconstructing original patterns upon reaggregation, and 4) genetic mosaics of morphogenetic mutations show local effects that may differ depending on the position of the mutant cells in the growing anlage.
Subject(s)
Body Patterning , Computer Simulation , Models, Biological , Algorithms , Animals , Body Patterning/genetics , Cell Division , Clone Cells , Gene Expression Regulation, Developmental , Mosaicism , Regeneration/physiology , Wings, Animal/growth & developmentABSTRACT
This paper reports a discussion between Antonio Garcia-Bellido and Lewis Wolpert about a number of questions raised by Alain Ghysen. The questions follow, in reverse order, the subjects dealt with in this issue: first the principles (are there unifying principles of development?), then questions dealing with evolution (why are patterns conserved?) and with the homeotic genes (what is their function?), then the cell biology of development (who is controlling actual morphogenesis?), and the generation and evolution of patterns (what makes development so reproducible and how does it change from one species to another?) and finally about the genetics of cell determination and specification (how does a cell measure its position?). Obviously the discussion did not provide any firm answers to any of these questions. Perhaps more importantly, it provides a vivid picture of two contrasting ways of thinking about developmental problems.