Regionalized tissue fluidization is required for epithelial gap closure during insect gastrulation.

Many animal embryos pull and close an epithelial sheet around the ellipsoidal egg surface during a gastrulation process known as epiboly. The ovoidal geometry dictates that the epithelial sheet first expands and subsequently compacts. Moreover, the spreading epithelium is mechanically stressed and this stress needs to be released. Here we show that during extraembryonic tissue (serosa) epiboly in the insect Tribolium castaneum, the non-proliferative serosa becomes regionalized into a solid-like dorsal region with larger non-rearranging cells, and a more fluid-like ventral region surrounding the leading edge with smaller cells undergoing intercalations. Our results suggest that a heterogeneous actomyosin cable contributes to the fluidization of the leading edge by driving sequential eviction and intercalation of individual cells away from the serosa margin. Since this developmental solution utilized during epiboly resembles the mechanism of wound healing, we propose actomyosin cable-driven local tissue fluidization as a conserved morphogenetic module for closure of epithelial gaps.

Morphogenesis of the ovarian follicular epithelium during initial stages of embryogenesis of the viviparous earwig, Hemimerus talpoides.

Representatives of the highly specialized earwig family Hemimeridae are epizoic and viviparous. Their embryos develop inside terminal ovarian follicles (termed also embryonic follicles) and rely solely on nutrients transferred from mother tissues. In this report, we present results of ultrastructural and histochemical studies of the initial stage of Hemimerus talpoides development. Our results show that the follicular cells surrounding fully grown oocyte of Hemimerus do not degenerate after initiation of embryogenesis, but transform and gradually form the wall of the incubation chamber in which the embryo develops. We also show that amniotic cells of the early embryo remain in direct contact with transformed follicular cells. In the region of contact, short outgrowths of the amniotic cells associate with irregular surface specializations of the transformed follicular cells. We suggest that extended "postfertilization" activity of hemimerid follicular cells represents an adaptation to viviparity and matrotrophy in this insect lineage.

A dataset of egg size and shape from more than 6,700 insect species.

Offspring size is a fundamental trait in disparate biological fields of study. This trait can be measured as the size of plant seeds, animal eggs, or live young, and it influences ecological interactions, organism fitness, maternal investment, and embryonic development. Although multiple evolutionary processes have been predicted to drive the evolution of offspring size, the phylogenetic distribution of this trait remains poorly understood, due to the difficulty of reliably collecting and comparing offspring size data from many species. Here we present a dataset of 10,449 morphological descriptions of insect eggs, with records for 6,706 unique insect species and representatives from every extant hexapod order. The dataset includes eggs whose volumes span more than eight orders of magnitude. We created this dataset by partially automating the extraction of egg traits from the primary literature. In the process, we overcame challenges associated with large-scale phenotyping by designing and employing custom bioinformatic solutions to common problems. We matched the taxa in this dataset to the currently accepted scientific names in taxonomic and genetic databases, which will facilitate the use of these data for testing pressing evolutionary hypotheses in offspring size evolution.

Organelle assemblages implicated in the transfer of oocyte components to the embryo: an insect perspective.

Besides reserve materials (yolk spheres, lipid droplets), ribosomes and various mRNA species, insect oocytes contain large easily morphologically recognizable organelle assemblages: the Balbiani body and the oosome (pole plasm). These assemblages are implicated in the transfer of oocyte components (mitochondria, polar granules) to the embryo that is to offspring. Here, we review present knowledge of morphology, morphogenesis, molecular composition and function/s of these assemblages. We discuss also the morphogenesis and presumed function of unconventional organelle assemblages, dormant stacks of endoplasmic reticulum, recently described in the oocytes and early embryos of a viviparous dermapteran, Hemimerus talpoides.

Evolution of maternal control of axial patterning in insects.

Positional and cell fate cues provided maternally to eggs are important factors in the development of many animals. The insects are a model clade where maternal establishment of embryonic axes is widespread and has been a topic of intense classical and molecular embryological analysis. Recently, significant progress has been made in revealing the molecular basis of some classical embryological experiments. In addition, observations of novel forms of maternal positional cues have been made. Finally, it has become increasingly clear that no maternal source of positional information acts alone without input and feedback from zygotic target genes to ensure precise and repeatable pattern formation in the early embryo. These advances will be discussed in the context of historical experiments, our current understanding of how positional cues can be generated, stored, and transmitted in insect ovaries and eggs, and how the nature of the cues can change in evolution.

Unusual morphological adaptations and processes associated with viviparity in an epizoic dermapteran.

Matrotrophic viviparity is a reproductive pattern in which offspring develop inside a female's body which provides gas exchange and nutrients necessary for development. Besides placental mammals, structural and physiological aspects of matrotrophic viviparity are poorly characterized. In insects, the majority of species is oviparous, i.e. lay eggs, and viviparous reproduction has been reported only in 11 out of 44 orders, including earwigs (Dermaptera). Among dermapterans, matrotrophic viviparity has been reported in two epizoic subgroups: Arixeniidae and Hemimeridae. Here, we provide morphological evidence for distinct adaptations for this mode of viviparity in embryonic and maternal tissues in a representative of the latter subgroup, Hemimerus talpoides. Our study reveals a novel mechanism of maternal contribution to embryonic development which operates during oogenesis and involves characteristic modification of endoplasmic reticulum cisternae. Conspicuous and apparently inactive para-crystalline stacks of the endoplasmic reticulum are deposited in the oocyte cytoplasm and become activated during early embryonic development. Our analyses indicate additionally that in Hemimerus, transformed follicular/ovarian cells (on the mother's side) and an evagination of the dorsal vessel (on the embryo's side) converge to form a cephalic vesicle, structure analogous to a placenta. The cellular architecture of this unusual "cephalic placenta" points to its participation in an exchange of low molecular weight substances between a mother and developing embryo.

The Function and Evolution of Nuclear Receptors in Insect Embryonic Development.

Nuclear receptors are a family of transcription factors that are often responsive to small ligands, allowing for efficient gene expression-level responses to a stimulus. The average insect has 21 genes encoding nuclear receptors, whose functions are especially well studied in developmental transitions during the insect life cycle, such as metamorphosis and molting. However, their utility as well-controlled transcriptional regulators also lends them to important roles in embryogenesis, neurogenesis, metabolism, and organogenesis. Such developmental functions have been explored in depth in the model organism Drosophila melanogaster. More recently, advances in genomic resources and functional genomic methodologies have allowed for comparison of nuclear receptor function among a wider range of insect species. As has been the trend throughout the field of Evo-Devo, these new data sets reveal that many genes are shared, but the ways in which they are utilized in different lineages are more variable. In this chapter, we describe the suite of nuclear receptor genes found in Drosophila and their developmental functions. We then compare and contrast these genes and their functions in diverse insects.

Neuronal migration during development and the amyloid precursor protein.

The Amyloid Precursor Protein (APP) is the source of amyloid peptides that accumulate in Alzheimer's disease. However, members of the APP family are strongly expressed in the developing nervous systems of invertebrates and vertebrates, where they regulate neuronal guidance, synaptic remodeling, and injury responses. In contrast to mammals, insects express only one APP ortholog (APPL), simplifying investigations into its normal functions. Recent studies have shown that APPL regulates neuronal migration in the developing insect nervous system, analogous to the roles ascribed to APP family proteins in the mammalian cortex. The comparative simplicity of insect systems offers new opportunities for deciphering the signaling mechanisms by which this enigmatic class of proteins contributes to the formation and function of the nervous system.

Embryonic axon guidance: insights from Drosophila and other insects.

During embryonic development, growing axons are guided by cellular signaling pathways that control a series of individual axon guidance decisions. In Drosophila, two major pathways (Netrin-Frazzled/DCC and Slit-Robo) regulate axon guidance in the embryonic ventral nerve cord, including the critical decision of whether or not to cross the midline. Studies in the fruit fly have revealed a complex picture of precise regulation and cross-talk between these pathways. In addition, Robo receptors in Drosophila have diversified their activities to regulate additional axon guidance decisions in the developing embryo. Here, I discuss recent advances in understanding roles and regulation of the Net-Fra and Slit-Robo signaling pathways in Drosophila, and examine the evolutionary conservation of these signaling mechanisms across insects and other arthropods.

Morphogenetic functions of extraembryonic membranes in insects.

Morphogenetic functions of the amnioserosa, the serosa, the amnion, and the yolk sac are reviewed on the basis of recent studies in flies (Drosophila, Megaselia), beetles (Tribolium), and hemipteran bugs (Oncopeltus). Three hypotheses are presented. First, it is suggested that the amnioserosa of Drosophila and the dorsal amnion of other fly species function in a similar manner. Second, it is proposed that in many species with an amniotic cavity, the amnion determines the site of serosa rupture, which, through interactions between the serosa and the amnion, enables the embryo to break free from the amniotic cavity and to close its backside. Finally, it is concluded that the yolk sac is likely an important player in insect morphogenesis.

Mechanisms of animal diapause: recent developments from nematodes, crustaceans, insects, and fish.

Life cycle delays are beneficial for opportunistic species encountering suboptimal environments. Many animals display a programmed arrest of development (diapause) at some stage(s) of their development, and the diapause state may or may not be associated with some degree of metabolic depression. In this review, we will evaluate current advancements in our understanding of the mechanisms responsible for the remarkable phenotype, as well as environmental cues that signal entry and termination of the state. The developmental stage at which diapause occurs dictates and constrains the mechanisms governing diapause. Considerable progress has been made in clarifying proximal mechanisms of metabolic arrest and the signaling pathways like insulin/Foxo that control gene expression patterns. Overlapping themes are also seen in mechanisms that control cell cycle arrest. Evidence is emerging for epigenetic contributions to diapause regulation via small RNAs in nematodes, crustaceans, insects, and fish. Knockdown of circadian clock genes in selected insect species supports the importance of clock genes in the photoperiodic response that cues diapause. A large suite of chaperone-like proteins, expressed during diapause, protects biological structures during long periods of energy-limited stasis. More information is needed to paint a complete picture of how environmental cues are coupled to the signal transduction that initiates the complex diapause phenotype, as well as molecular explanations for how the state is terminated. Excellent examples of molecular memory in post-dauer animals have been documented in Caenorhabditis elegans It is clear that a single suite of mechanisms does not regulate diapause across all species and developmental stages.

Comparative embryogenesis of Mecoptera and Lepidoptera with special reference to the abdominal prolegs.

The eruciform larvae of holometabolous insects are primarily characterized by bearing a varying number of abdominal prolegs in addition to three pairs of thoracic legs. However, whether the prolegs are evolutionarily homologous among different insect orders is still a disputable issue. We examined the embryonic features and histological structure of the prolegs of the scorpionfly Panorpa byersi Hua and Huang (Mecoptera: Panorpidae) and the Oriental armyworm Mythimna separata (Walker) (Lepidoptera: Noctuidae) to investigate whether the prolegs are homologous between these two holometabolous insect orders. In the scorpionfly, paired lateral process primordia arise on abdominal segments I-VIII (A1-A8) in line with the thoracic legs in early embryonic stages, but degenerate into triangular protuberances in later stages, and paired medial processes appear along the midventral line before dorsal closure and eventually develop into unjointed, cone-shaped prolegs. Histological observation showed that the lumina of the prolegs are not continuous with the hemocoel, differing distinctly from that of the basic appendicular plan of thoracic legs. These results suggest that the prolegs are likely secondary outgrowths in Mecoptera. In the armyworm, lateral process primordia appear on A1-A10 in alignment with the thoracic legs in the early embryonic stages, although only the rudiments on A3-A6 and A10 develop into segmented prolegs with the lumina continuous with the hemocoel and others degenerate eventually, suggesting that the prolegs are true segmental appendages serially homologous with the thoracic legs in Lepidoptera. Therefore, we conclude that the larval prolegs are likely not evolutionarily homologous between Mecoptera and Lepidoptera.

Evolution of early male-killing in horizontally transmitted parasites.

Early male-killing (MK) bacteria are vertically transmitted reproductive parasites which kill male offspring that inherit them. Whereas their incidence is well documented, characteristics allowing originally non-MK bacteria to gradually evolve MK ability remain unclear. We show that horizontal transmission is a mechanism enabling vertically transmitted bacteria to evolve fully efficient MK under a wide range of host and parasite characteristics, especially when the efficacy of vertical transmission is high. We also show that an almost 100% vertically transmitted and 100% effective male-killer may evolve from a purely horizontally transmitted non-MK ancestor, and that a 100% efficient male-killer can form a stable coexistence only with a non-MK bacterial strain. Our findings are in line with the empirical evidence on current MK bacteria, explain their high efficacy in killing infected male embryos and their variability within and across insect taxa, and suggest that they may have evolved independently in phylogenetically distinct species.

A conserved genetic mechanism specifies deutocerebral appendage identity in insects and arachnids.

The segmental architecture of the arthropod head is one of the most controversial topics in the evolutionary developmental biology of arthropods. The deutocerebral (second) segment of the head is putatively homologous across Arthropoda, as inferred from the segmental distribution of the tripartite brain and the absence of Hox gene expression of this anterior-most, appendage-bearing segment. While this homology statement implies a putative common mechanism for differentiation of deutocerebral appendages across arthropods, experimental data for deutocerebral appendage fate specification are limited to winged insects. Mandibulates (hexapods, crustaceans and myriapods) bear a characteristic pair of antennae on the deutocerebral segment, whereas chelicerates (e.g. spiders, scorpions, harvestmen) bear the eponymous chelicerae. In such hexapods as the fruit fly, Drosophila melanogaster, and the cricket, Gryllus bimaculatus, cephalic appendages are differentiated from the thoracic appendages (legs) by the activity of the appendage patterning gene homothorax (hth). Here we show that embryonic RNA interference against hth in the harvestman Phalangium opilio results in homeonotic chelicera-to-leg transformations, and also in some cases pedipalp-to-leg transformations. In more strongly affected embryos, adjacent appendages undergo fusion and/or truncation, and legs display proximal defects, suggesting conservation of additional functions of hth in patterning the antero-posterior and proximo-distal appendage axes. Expression signal of anterior Hox genes labial, proboscipedia and Deformed is diminished, but not absent, in hth RNAi embryos, consistent with results previously obtained with the insect G. bimaculatus. Our results substantiate a deep homology across arthropods of the mechanism whereby cephalic appendages are differentiated from locomotory appendages.

Multispecies Analysis of Expression Pattern Diversification in the Recently Expanded Insect Ly6 Gene Family.

Gene families often consist of members with diverse expression domains reflecting their functions in a wide variety of tissues. However, how the expression of individual members, and thus their tissue-specific functions, diversified during the course of gene family expansion is not well understood. In this study, we approached this question through the analysis of the duplication history and transcriptional evolution of a rapidly expanding subfamily of insect Ly6 genes. We analyzed different insect genomes and identified seven Ly6 genes that have originated from a single ancestor through sequential duplication within the higher Diptera. We then determined how the original embryonic expression pattern of the founding gene diversified by characterizing its tissue-specific expression in the beetle Tribolium castaneum, the butterfly Bicyclus anynana, and the mosquito Anopheles stephensi and those of its duplicates in three higher dipteran species, representing various stages of the duplication history (Megaselia abdita, Ceratitis capitata, and Drosophila melanogaster). Our results revealed that frequent neofunctionalization episodes contributed to the increased expression breadth of this subfamily and that these events occurred after duplication and speciation events at comparable frequencies. In addition, at each duplication node, we consistently found asymmetric expression divergence. One paralog inherited most of the tissue-specificities of the founder gene, whereas the other paralog evolved drastically reduced expression domains. Our approach attests to the power of combining a well-established duplication history with a comprehensive coverage of representative species in acquiring unequivocal information about the dynamics of gene expression evolution in gene families.

The embryonic development of Stylops ovinae (Strepsiptera, Stylopidae) with emphasis on external morphology.

External features of the embryonic development of Stylops ovinae (Strepsiptera) were examined. Eighteen distinct embryological stages are suggested. Many embryological traits are closely correlated to the parasitic life style of the first instar larvae or to vivipary. The high number of eggs, their small size, the characteristic egg membrane, and the lack of micropyles are derived groundplan features of Strepsiptera. The development with a semi-long germ embryo is shared with several other groups of Holometabola. The reduction of the labrum and antennae are autapomorphies of Strepsiptera. The cephalic ventral plate of the first instar larva of S. ovinae is formed by parts of the head capsule and the anlagen of the maxillae and labium. It is involved in the formation of the specific entognathous condition, and the entire character complex is autapomorphic for Stylopidae. The trochanter is recognizable in the anlagen of all three legs. Its fusion with the femur in the later stages is an autapomorphy of Stylopidia. The extreme spiralization and compression of the abdomen during blastokinesis is a derived feature, like the reduction of the anlagen of the anterior abdominal appendages. The caudal bristles on segment XI are possibly re-activated cerci. The same is likely in the case of segment XI.

The structure and ultrastructure of the egg capsule of Brachyptera risi (Plecoptera, Nemouroidea, Taeniopterygidae) with some remarks concerning choriogenesis.

The organization of the egg capsule of the euholognathan stonefly, which represents the family Taeniopterygidae (Nemuroidea) was investigated using light and electron microscopy techniques. The presence of a complex, multilayered egg capsule, composed of a vitelline envelope, multilayered chorion, and extrachorion is described. The morphology of the eggshell of Brachyptera risi was compared with that of euholognthan and systellognathan egg coverings and the ground plan of the egg capsule in Plecoptera was discussed.

[Effects of the Vital Activity of Soil Insect Larvae on Microbial Processes in the Soil].

The effects of Elateridae larvae (wireworms) on the structure, functional diversity, and tolerance of the soil microbial population in steppe ecosystems have been investigated. The trophic and locomotor activity of wireworms leads to an appreciable increase in bacterial abundance and suppression of fungal activity. The fungal hyphae in the presence of wireworms are significantly damaged, which can be related to the feeding activity of Elateridae. The increase of bacterial abundance on the background of exclusion of the fungal component shifts the microbial succession to the acceleration of organic matter mineralization. The microbial consumption of mono- and oligosaccharides, alcohols, and water-soluble compounds increases in the presence of wireworms (multisubstrate test). The effect of Elateridae larvae on the microorganisms transforming nitrogen compounds is species-specific. Agriotes obscurus activity decreases their consumption of urea and creatinine by 2.1-2.5 times, and Selatosomus aeneus increases it by 1.3 and 2.5 times, respectively. The intensity of actual nitrogen fixation in the soil increases in the presence of wireworms by almost 4 times, but the losses of gaseous nitrogen do not increase because of the decrease in both the denitrification and methanogenesis rates

The development and functions of oenocytes.

Oenocytes have intrigued insect physiologists since the nineteenth century. Many years of careful but mostly descriptive research on these cells highlights their diverse sizes, numbers, and anatomical distributions across Insecta. Contemporary molecular genetic studies in Drosophila melanogaster and Tribolium castaneum support the hypothesis that oenocytes are of ectodermal origin. They also suggest that, in both short and long germ-band species, oenocytes are induced from a Spalt major/Engrailed ectodermal zone by MAPK signaling. Recent glimpses into some of the physiological functions of oenocytes indicate that they involve fatty acid and hydrocarbon metabolism. Genetic studies in D. melanogaster have shown that larval oenocytes synthesize very-long-chain fatty acids required for tracheal waterproofing and that adult oenocytes produce cuticular hydrocarbons required for desiccation resistance and pheromonal communication. Exciting areas of future research include the evolution of oenocytes and their cross talk with other tissues involved in lipid metabolism such as the fat body.