Leg development in flies versus grasshoppers: differences in dpp expression do not lead to differences in the expression of downstream components of the leg patterning pathway.
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All insect legs are structurally similar, characterized by five primary segments. However, this final form is achieved in different ways. Primitively, the legs developed as direct outgrowths of the body wall, a condition retained in most insect species. In some groups, including the lineage containing the genus Drosophila, legs develop indirectly from imaginal discs. Our understanding of the molecular mechanisms regulating leg development is based largely on analysis of this derived mode of leg development in the species D. melanogaster. The current model for Drosophila leg development is divided into two phases, embryonic allocation and imaginal disc patterning, which are distinguished by interactions among the genes wingless (wg), decapentaplegic (dpp) and distalless (dll). In the allocation phase, dll is activated by wg but repressed by dpp. During imaginal disc patterning, dpp and wg cooperatively activate dll and also indirectly inhibit the nuclear localization of Extradenticle (Exd), which divide the leg into distal and proximal domains. In the grasshopper Schistocerca americana, the early expression pattern of dpp differs radically from the Drosophila pattern, suggesting that the genetic interactions that allocate the leg differ between the two species. Despite early differences in dpp expression, wg, Dll and Exd are expressed in similar patterns throughout the development of grasshopper and fly legs, suggesting that some aspects of proximodistal (P/D) patterning are evolutionarily conserved. We also detect differences in later dpp expression, which suggests that dpp likely plays a role in limb segmentation in Schistocerca, but not in Drosophila. The divergence in dpp expression is surprising given that all other comparative data on gene expression during insect leg development indicate that the molecular pathways regulating this process are conserved. However, it is consistent with the early divergence in developmental mode between fly and grasshopper limbs. (+info)
A cell adhesion protein from the crayfish Pacifastacus leniusculus, a serine proteinase homologue similar to Drosophila masquerade.
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A cDNA encoding a protein resembling masquerade, a serine proteinase homologue expressed during embryogenesis, larval, and pupal development in Drosophila melanogaster, was identified in hemocytes of the adult freshwater crayfish, Pacifastacus leniusculus. The crayfish protein is similar to Drosophila masquerade in the following aspects: (a) overall sequence of the serine proteinase domain, such as the position of three putative disulfide bridges, glycine in the place of the catalytic serine residue, and the presence of a substrate-lining pocket typical for trypsins; (b) the presence of several copies of a disulfide-knotted motif in the putative propeptide. This masquerade-like protein is cleaved into a 27-kDa fragment, which could be detected by immunoblot analysis using an affinity-purified antibody against a synthetic peptide in the C-terminal domain of the protein. The 27-kDa protein could be immunoaffinity-purified from hemocyte lysate supernatant and exhibited cell adhesion activity in vitro, indicating that the C-terminal domain of the crayfish masquerade-like protein mediates cell adhesion. (+info)
Transmission thresholds for dengue in terms of Aedes aegypti pupae per person with discussion of their utility in source reduction efforts.
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The expense and ineffectiveness of drift-based insecticide aerosols to control dengue epidemics has led to suppression strategies based on eliminating larval breeding sites. With the notable but short-lived exceptions of Cuba and Singapore, these source reduction efforts have met with little documented success; failure has chiefly been attributed to inadequate participation of the communities involved. The present work attempts to estimate transmission thresholds for dengue based on an easily-derived statistic, the standing crop of Aedes aegypti pupae per person in the environment. We have developed these thresholds for use in the assessment of risk of transmission and to provide targets for the actual degree of suppression required to prevent or eliminate transmission in source reduction programs. The notion of thresholds is based on 2 concepts: the mass action principal-the course of an epidemic is dependent on the rate of contact between susceptible hosts and infectious vectors, and threshold theory-the introduction of a few infectious individuals into a community of susceptible individuals will not give rise to an outbreak unless the density of vectors exceeds a certain critical level. We use validated transmission models to estimate thresholds as a function of levels of pre-existing antibody levels in human populations, ambient air temperatures, and size and frequency of viral introduction. Threshold levels were estimated to range between about 0.5 and 1.5 Ae. aegypti pupae per person for ambient air temperatures of 28 degrees C and initial seroprevalences ranging between 0% to 67%. Surprisingly, the size of the viral introduction used in these studies, ranging between 1 and 12 infectious individuals per year, was not seen to significantly influence the magnitude of the threshold. From a control perspective, these results are not particularly encouraging. The ratio of Ae. aegypti pupae to human density has been observed in limited field studies to range between 0.3 and >60 in 25 sites in dengue-endemic or dengue-susceptible areas in the Caribbean, Central America, and Southeast Asia. If, for purposes of illustration, we assume an initial seroprevalence of 33%, the degree of suppression required to essentially eliminate the possibility of summertime transmission in Puerto Rico, Honduras, and Bangkok, Thailand was estimated to range between 10% and 83%; however in Mexico and Trinidad, reductions of >90% would be required. A clearer picture of the actual magnitude of the reductions required to eliminate the threat of transmission is provided by the ratio of the observed standing crop of Ae. aegypti pupae per person and the threshold. For example, in a site in Mayaguez, Puerto Rico, the ratio of observed and threshold was 1.7, meaning roughly that about 7 of every 17 breeding containers would have to be eliminated. For Reynosa, Mexico, with a ratio of approximately 10, 9 of every 10 containers would have to be eliminated. For sites in Trinidad with ratios averaging approximately 25, the elimination of 24 of every 25 would be required. With the exceptions of Cuba and Singapore, no published reports of sustained source reduction efforts have achieved anything near these levels of reductions in breeding containers. Practical advice on the use of thresholds is provided for operational control projects. (+info)
atonal regulates neurite arborization but does not act as a proneural gene in the Drosophila brain.
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Drosophila atonal (ato) is the proneural gene of the chordotonal organs (CHOs) in the peripheral nervous system (PNS) and the larval and adult photoreceptor organs. Here, we show that ato is expressed at multiple stages during the development of a lineage of central brain neurons that innervate the optic lobes and are required for eclosion. A novel fate mapping approach shows that ato is expressed in the embryonic precursors of these neurons and that its expression is reactivated in third instar larvae (L3). In contrast to its function in the PNS, ato does not act as a proneural gene in the embryonic brain. Instead, ato performs a novel function, regulating arborization during larval and pupal development by interacting with Notch. (+info)
Identification and developmental expression of inhibitor of caspase-activated DNase (ICAD) in Drosophila melanogaster.
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DNA fragmentation, a hallmark of apoptosis, is regulated by a specific nuclease called caspase-activated DNase (CAD) and its inhibitor (ICAD). When cell lysates from Drosophila S2 cells were chemically denatured and the denatured proteins were removed after dialysis, the supernatant inhibited Drosophila CAD (dCAD). To identify the inhibitor, we tested recombinant DREP-1, which was previously identified using the Drosophila EST data base and found it also inhibited dCAD DNase. An antibody against DREP-1 inhibited the ICAD activity in the S2 cell extracts, confirming the identification of DREP-1 as a Drosophila homolog of ICAD (dICAD). The recombinant DREP-1/dICAD was cleaved at a specific site by human caspase 3 as well as by extracts prepared from S2 cells undergoing apoptosis. Biochemical fractionation and immunoprecipitation of dICAD from S2 cell extracts indicated that dICAD is complexed with dCAD in proliferating cells. The expression of the caspase-resistant form of dICAD/DREP-1 in a Drosophila neuronal cell line prevented the apoptotic DNA fragmentation. Northern hybridization and the immunohistochemical analyses revealed that the expression of the dICAD gene is developmentally regulated. (+info)
A transient specialization of the microtubule cytoskeleton is required for differentiation of the Drosophila visual system.
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Drosophila beta3-tubulin is an essential isoform expressed during differentiation of many cell types in embryos and pupae. We report here that during pupal development transient beta3 expression demarcates a unique subset of neurons in the developing adult visual system. beta3 is coassembled into microtubules with beta1, the sole beta-tubulin isoform in the permanent microtubule cytoskeleton of the adult eye and brain. Examination of beta3 mutant phenotypes showed that beta3 is required for axonal patterning and connectivity and for spatial positioning within the optic lobe. Comparison of the phenotypes of beta3 mutations with those that result from disruption of the Hedgehog signaling pathway shows that beta3 functions early in the establishment of the adult visual system. Our data support the hypothesis that beta3 confers specialized properties on the microtubules into which it is incorporated. Thus a transient specialization of the microtubule cytoskeleton during differentiation of a specific subset of the neurons has permanent consequences for later cell function. (+info)
Genetic and molecular complexity of the position effect variegation modifier mod(mdg4) in Drosophila.
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mod(mdg4), also known as E(var)3-93D, is involved in a variety of processes, such as gene silencing in position effect variegation (PEV), the control of gypsy insulator sequences, regulation of homeotic gene expression, and programmed cell death. We have isolated a large number of mod(mdg4) cDNAs, representing 21 different isoforms generated by alternative splicing. The deduced proteins are characterized by a common N terminus of 402 amino acids, including the BTB/POZ-domain. Most of the variable C termini contain a new consensus sequence, including four positioned hydrophobic amino acids and a Cys(2)His(2) motif. Using specific antibodies for two protein isoforms, we demonstrate different distributions of the corresponding proteins on polytene chromosomes. Mutations in the genomic region encoding exons 1-4 show enhancement of PEV and homeotic transformation and affect viability and fertility. Homeotic and PEV phenotypes are enhanced by mutations in other trx-group genes. A transgene containing the common 5' region of mod(mdg4) that is present in all splice variants known so far partially rescues the recessive lethality of mod(mdg4) mutant alleles. Our data provide evidence that the molecular and genetic complexity of mod(mdg4) is caused by a large set of individual protein isoforms with specific functions in regulating the chromatin structure of different sets of genes throughout development. (+info)
Imaginal pioneers prefigure the formation of adult thoracic muscles in Drosophila melanogaster.
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In insects, specialized mesodermal cells serve as templates to organize myoblasts into distinct muscle fibers during embryogenesis. In the grasshopper embryo, large mesodermal cells called muscle pioneers extend between the epidermal attachment points of future muscle fibers and serve as foci for myoblast fusion. In the Drosophila embryo, muscle founder cells serve a similar function, organizing large numbers of myoblasts into larval muscles. During the metamorphosis of Drosophila, nearly all larval muscles degenerate and are replaced by a set of de novo adult muscles. The extent to which specialized mesodermal cells homologous to the founders and pioneers of the insect embryo are involved in the development of adult-specific muscles has yet to be established. In the larval thorax, the majority of imaginal myoblasts are associated with the imaginal discs. We report here the identification of a morphologically distinct class of disc-associated myoblasts, which we call imaginal pioneers, that prefigures the formation of at least three adult-specific muscles, the tergal depressor of the trochanter and dorsoventral muscles I and II. Like the muscle pioneers of the grasshopper, the imaginal pioneers attach to the epidermis at sites where the future muscle insertions will arise and erect a scaffold for developing adult muscles. These findings suggest that a prior segregation of imaginal myoblasts into at least two populations, one of which may act as pioneers or founders, must occur during development. (+info)