Formation and structure of nutritive cords in telotrophic ovarioles of snake flies (Insecta: Raphidioptera)

Telotrophic ovariole of Raphidia spp. is composed of the anteriorly located terminal filament, tube-shaped tropharium, the vitellarium and the ovariole stalk. The tropharium consists of a central syncytial core surrounded by one cell thick layer of tapetum cells. Early previtellogenic oocytes differentiate at the base of tropharium. Both the oocytes and the tapetum cells are connected with the central syncytium by delicate intercellular bridges. At the onset of previtellogenic growth, the anterior parts of the oocytes become extended and form long cytoplasmic projections--nutritive cords. Each nutritive cord contains numerous microtubules that show no preferential orientation within the cord but diminishing anterior-posterior gradient of distribution. Irregular arrangement of microtubules indicates that this cytoskeletal scaffold does not play any role in directed transport within the ovariole but instead constitutes one of the elements of the structural framework of the nutritive cord. Besides microtubules, the stability of the nutritive cords in Raphidia ovarioles is maintained by the rim-shaped membrane foldings lined with microfilaments.     

The larval head of Raphidia (Raphidioptera, Insecta) and its phylogenetic significance

External and internal head structures of larval representatives of Raphidiidae are described. The obtained data were compared to characters of other neuropterid larvae and to larval characters of representatives of other endopterygote lineages. Characters potentially relevant for phylogenetic reconstruction are listed and discussed. The larvae of Raphidioptera differ distinctly from other neuropterid larvae in their morphology. They are mainly characterised by autapomorphic and plesiomorphic character states and few features indicate systematic affinities with other groups. Endopterygote groundplan features maintained in Raphidioptera are the complete tentorium, the free labrum, the full set of labral muscles, the presence of four extrinsic antennal muscles, the three-segmented labial palpi, the presence of a full set of extrinsic maxillary and labial muscles, the presence of a salivarium, and possibly the high number of stemmata. Apomorphies likely correlated with predaceous habits are the long gula, the protracted maxillae, the longitudinal arrangement of extrinsic maxillary muscles, and the elongated prepharyngeal tube. Highly unusual, potentially autapomorphic features are the presence of a dorsal ligament of the tentorium and paired gland-like structures below the pharynx. A prognathous or very slightly inclined head and slender mandibles without mola are features shared by larvae of all orders of Neuropterida. The parallel-sided head is a potential synapomorphy of Raphidioptera and Megaloptera. A fully prognathous head with anteriorly shifted posterior tentorial grooves and the presence of a parietal ridge and a distinct neck region are features shared with Corydalidae. Characters of the larval head are not sufficient for a reliable placement of Raphidioptera.     

New snakeflies (Insecta: Raphidioptera) from the Lower Cretaceous of the UK,Spain and Brazil

Abstract: Several new taxa of snakeflies (Raphidioptera) are described from the Lower Cretaceous deposits of the Wealden, UK (Barremian), Montsec, Spain (Barremian) and the Crato Formation, Brazil (Aptian). Mesoraphidia ednae sp. nov. and M. hilli sp. nov. are described from the Wealden; Nanoraphidia lithographica sp. nov. and Iberoraphidia dividua gen. et sp. nov. are described from Montsec, and Baissoptera lisae sp. nov. is described from the Crato Formation. The geographical range of Nanoraphidia has potentially been extended.     

The larval development of the telotrophic meroistic ovary in the bug Dysdercus intermedius (Heteroptera, Pyrrhocoridae)

Bug ovaries are of the telotrophic meroistic type. Nurse cells are restricted to the anterior tropharium and are in syncytial connection with the oocytes via the acellular trophic core region into which cytoplasmic projections of oocytes and nurse cells open. The origin of intercellular connections in bug ovaries is not well understood. In order to elucidate the cellular processes underlying the emergence of the syncytium, we analysed the development of the ovary of Dysdercus intermedius throughout the five larval instars. Up to the third instar, the germ cell population of an ovariole anlage forms a single, tight rosette. In the center of the rosette, phosphotyrosine containing proteins and f-actin accumulate. This center is filled with fusomal cytoplasm and closely interdigitating cell membranes known as the membrane labyrinth. With the molt to the fourth instar germ cells enhance their mitotic activity considerably. As a rule, germ cells divide asynchronously. Simultaneously, the membrane labyrinth expands and establishes a central column within the growing tropharium. In the fifth instar the membrane labyrinth retracts to an apical position, where it is maintained even in ovarioles of adult females. The former membrane labyrinth in middle and posterior regions of the tropharium is replaced by the central core to which nurse cells and oocytes are syncytially connected. Germ cells in the most anterior part of the tropharium, i.e. those in close proximity to the membrane labyrinth remain proliferative. The posterior-most germ cells enter meiosis and become oocytes. The majority of the ovarioles' germ cells, located in between these two populations, endopolyploidize and function as nurse cells. We conclude that the extensive multiplication of germ cells and their syncytial assembly during larval development is achieved by incomplete cytokineses followed by massive membrane production. Membranes are degraded as soon as the trophic core develops. For comparative reasons, we also undertook a cursory examination of early germ cell development in Dysdercus intermedius males. All results were compatible with the known basic patterns of early insect spermatogenesis. Germ cells run through mitotic and meiotic divisions in synchronous clusters emerging from incomplete cytokineses. During the division phase, the germ cells of an individual cluster are connected by a polyfusome rich in f-actin.     

The ovary ofCatajapyx aquilonaris (Insecta,Entognatha): ultrastructure of germarium and terminal filament

Summary Each of the two ovaries ofCatajapyx aquilonaris is composed of seven segmentally (metamerically) arranged ovarioles. The two lateral oviducts that join and bear ovarioles extend throughout the abdomen. In the ovariole three regions can be recognized: the terminal filament, the germarium and the vitellarium. The terminal filaments do not fuse with each other but attach separately (by means of muscle fibres) to the closest lobes of the fat body. Germ cells in the germarium are not joined by intercellular bridges and do not form clusters. Thus the ovarioles ofC. aquilonaris are interpreted as being primarily panoistic. The results obtained support the hypothesis that both dipluran subgroups (Campodeina and Japygina) do not form a monophyletic unit.     

Development and cellular differentiation of an insect telotrophic ovary (Rhodnius prolixus)

Differentiation events accompanying the larval-adult ovarian transformation in Rhodnius prolixus can be divided into three phases: proliferative phase (unfed to 3 days post-feed or DPF), early differentiation phase (9–15 DPF) and late differentiation phase (16 DPF to moult at 21 DPF). Ovarioles remain morphologically larval until feeding initiates development. The unfed ovariole contains germ cells surrounding a central trophic core region with the ‘germarial lumen’ occupying the basal region of the tropharium immediately above the pre-follicular tissue. Mitosis of germ cells during the proliferative phase results in a progressive increase in tropharial size with no differentiation of tissues. Regional specialization within the ovariole marks the beginning of the early differentiation phase. A zone of oocytes is established at the base of the tropharium with nuclei containing synaptonemal complexes and condensing chromosomes. Nurse cell differentiation is characterized by nucleolar elaboration and nucleo-cytoplasmic transport, the cytoplasm becoming rich in ribosomes. Autoradiographic results suggest that functional nurse cell-oocyte divergence occurs concurrently with morphological divergence. Pre-follicular tissue is divided into apical and basal zones with apical zone differentiation occurring during early and late differentiation phases.     

Development of nurse cell-oocyte interactions in the insect telotrophic ovary (Rhodnius prolixus)

The establishment and reorganization of intercellular bridges during larval-adult ovarian differentiation is the basis of the syncytial nature of the adult hemipteran telotrophic ovary. The formation, in the late differentiation phase, of groups of closely arranged nurse cell nuclei occupying a common cytoplasm results from membrane fusions. Oocyte-oocyte intercellular bridge systems later are modified to form the trophic cords. The trophic core, which undergoes a restructuring during the late differentiation phase, mediates nurse cell-oocyte interactions in this system. Material, transported to and accumulated by late differentiation phase pre-vitellogenic oocytes, originates from trophic core restructuring and zone III nurse cell production.     

Ultrastructural studies of the ovary of Palaeococcus fuscipennis (Burmeister) (Insecta, Hemiptera, Coccinea: Monophlebidae)

Ovaries of Palaeocoocus fuscipennis are composed of about 100 telotrophic ovarioles that are devoid of terminal filaments. In the ovariole a tropharium ( = trophic chamber) and vitellarium can be distinguished. The tropharium contains 7 trophocytes. A single oocyte develops in the vitellarium. The oocyte is surrounded by follicular cells that do not undergo diversification into subpopulations. The obtained results are discussed in a phylogenetic context.     

Bacteriocyte-like cells harbour Wolbachia in the ovary of Drosophila melanogaster (Insecta, Diptera) and Zyginidia pullula (Insecta, Hemiptera)

Wolbachia is the most widespread bacterial endosymbiont in insects. It is responsible for a variety of reproductive alterations of the hosts. Wolbachia is transmitted through the germline from mother to offspring and, in rare cases, between individuals. This implies that acquired properties (through symbiosis with Wolbachia) can become heritable. We investigated the transovarial inheritance of Wolbachia in two phylogenetically distant insects, Drosophila melanogaster and Zyginidia pullula. We detected in both systems bacteriocyte-like cells, densely packed with Wolbachia endosymbionts, at the tip of the ovarioles. Bacteriocytes are cells specialized to harbour bacteria, typical of mutualistic insect symbiosis. Our observations of bacteriocyte-like cells harbouring Wolbachia in the ovary emphasize the plasticity of the female reproductive system of insects, which maintains its function while some cells are densely colonized by bacteria. In summary, there is evidence from different insects that bacteria which behave as parasites of reproduction are harboured by cells resembling bacteriocytes, which appear to mediate transmission of the bacteria to the progeny. It seems a valid hypothesis that the bacteriocyte-like cells that we observed are not the result of a co-evolution of host and symbiont, considering that Wolbachia is not an obligatory symbiont in Drosophila and Zyginidia.     

Germ cell cluster in the panoistic ovary of Thysanoptera (Insecta)

Summary Germ cell clusters are found in the germarial region of ovarioles of Parthenothrips dracenae. Cluster mitoses are synchronized, at least initially. The intercellular bridges are filled with fusomal material, which can fuse to form polyfusomal aggregates which in turn form small rosettes. All cells develop into oocytes. Oocytes become isolated by a secondary detachment process. Intercellular bridges, together with fusomal material and cell membranes, survive for some time as isolated bodies. Phylogenetic consequences are discussed. The data provide strong evidence for a secondary panoistic ovary in thysanopterans, since cluster formation in ovaries of primary panoists has not been shown.     

Symmetrical pattern of follicle arrangement in the ovary of Musca domestica (Insecta,Diptera)

Summary The position of the oocyte nucleus within the ooplasm is fixed during the mid and late stages of house fly oogenesis. The germinal vesicle is located near the border of the nurse chamber, towards the periphery of the oocyte. The position of the anlage of the chorion raphe is strictly related to the germinal vesicle. As the raphe corresponds to the dorsal side of the later embryo, both the position of the oocyte nucleus and the raphe anlage in the follicular epithelium are early indicators of the dorsoventral axis of the house fly egg cell. In cross sections of the ovary the follicles are arranged in several concentric circles. The dorsal sides of all follicles within the ovary are oriented to an imaginary center. This center of orientation lies eccentrically near the medial part of the female abdomen. The resulting symmetrical pattern can be observed throughout the course of oogenesis. This implies that only a few follicles have the same dorsoventral orientation as the mother fly, and therefore this arrangement is contradictory to the imprinting hypotheses of body axis formation as well as to a possible inductive role of gravity.Supported by the Deutsche Forschungsgemeinschaft     

The development of microtubular arrays in the germ tissue of an insect telotrophic ovary

The microtubular arrays characteristic of the trophic core and cords of the adult Rhodnius prolixus ovary develop prior and during the larval--adult transformation. Development of the microtubules was revealed by immunocytochemistry, electron microscopy and polyacrylamide electrophoresis and Western blot analysis. Microtubular arrays were first detected in the trophic cords and presumptive trophic core 6 days before the adult molt. Cord microtubules increase in length and numbers as the trophic cords grow. Three microtubule packed cords have formed by 1 day post molt. The microtubule distribution in the presumptive core is non-uniform. Microtubule packed areas are interspersed with areas devoid of microtubules. The adult core begins forming between 1 day before molt and molting. This early adult core arises from the fusion of the anterior portions of microtubule packed cords. A fully mature adult core is not present by 2 days post molt. The microtubule packing density in the core increases from 2 days before to 2 days post molt. Tubulin increases from 6 days to 1 day before the adult molt.     

Male meiosis in camel-flies (Raphidioptera; Neuropteroidea)

Male meiosis in 3 species of the raphidioptera genus Agulla-- A. bicolor Banks, A. astuta (Banks), and A. bractea Carpenter-- closely parallels that of Neuroptera. The diploid complement in each comprises 12 pairs of autosomes plus X and Y; all are mediokinetic. One male of A. bicolor carried an extra pair of autosomes indistinguishable from the shortest member of the usual set: these formed a normal bivalent and segregated synchronously with the other autosomes. The spindle is formed by the collocation of individual units which envelope each chromosomal mass. The sex chromsomes are spatially separate on emergence from the joint vesicle of early prophase; oriented toward opposite poles they move into this interpolar axis and a central spindle unit forms about them. This unit elongates disproportionately in early premetaphase, and its subsequent contraction is not synchronous with that of the other units. Distance segregation of X and Y is completed in early premetaphase. Autosomal bivalents are chiasmate; their congressional maneuvers involve, in addition to the usual interpolar oscillations, a lateral movement to the periphery of the spindle to form a variably complete ring at the equator. Autosomal univalents occurs with a frequency of 13% in A. bicolor, 2% in A. astuta, and 1% in A. bractea; they undergo distance segregation with the sex chromosomes in the central spindle unit. The phylogenetic significance of the data is considered.     

Absence of ribosomal DNA amplification in the meroistic (telotrophic) ovary of the large milkweed bug Oncopeltus fasciatus (Dallas) (Hemiptera: Lygaeidae)

In the typical meroistic insect ovary, the oocyte nucleus synthesizes little if any RNA. Nurse cells or trophocytes actively synthesize ribosomes which are transported to and accumulated by the oocyte. In the telotrophic ovary a morphological separation exists, the nurse cells being localized at the apical end of each ovariole and communicating with the ooocytes via nutritive cords. In order to determine whether the genes coding for ribosomal RNA (rRNA) are amplified in the telotrophic ovary of the milkweed bug Oncopeltus fasciatus, the percentages of the genome coding for ribosomal RNA in somatic cells, spermatogenic cells, ovarian follicles, and nurse cells were compared. The oocytes and most of the nurse cells of O. fasciatus are uninucleolate. DNA hybridizing with ribosomal RNA is localized in a satellite DNA, the density of which is 1.712 g/cm(-3). The density of main-band DNA is 1.694 g/cm(-3). The ribosomal DNA satellite accounts for approximately 0.2% of the DNA in somatic and gametogenic tissues of both males and females. RNA-DNA hybridization analysis demonstrates that approximately 0.03% of the DNA in somatic tissues, testis, ovarian follicles, and isolated nurse cells hybridizes with ribosomal RNA. The fact that the percentage of DNA hybridizing with rRNA is the same in somatic and in male and female gametogenic tissues indicates that amplification of ribosomal DNA does not occur in nurse cells and that if it occurs in oocytes, it represents less than a 50-fold increase in ribosomal DNA. An increase in total genome DNA accounted by polyploidization appears to provide for increasing the amount of ribosomal DNA in the nurse cells.     

Snakefly diversity in Early Cretaceous amber from Spain (Neuropterida, Raphidioptera)

The Albian amber from Spain presently harbors the greatest number and diversity of amber adult fossil snakeflies (Raphidioptera). Within Baissopteridae, Baissoptera? cretaceoelectra sp. n., from the Peñacerrada I outcrop (Moraza, Burgos), is the first amber inclusion belonging to the family and described from western Eurasia, thus substantially expanding the paleogeographical range of the family formerly known from the Cretaceous of Brazil and eastern Asia. Within the family Mesoraphidiidae, Necroraphidia arcuata gen. et sp. n. and Amarantoraphidia ventolina gen. et sp. n. are described from the El Soplao outcrop (Rábago, Cantabria), whereas Styporaphidia? hispanica sp. n. and Alavaraphidia imperterrita gen. et sp. n. are describedfrom Peñacerrada I. In addition, three morphospecies are recognized from fragmentary remains. The following combinations are restored: Yanoraphidia gaoi Ren, 1995, stat. rest., Mesoraphidia durlstonensis Jepson, Coram and Jarzembowski, 2009, stat. rest., and Mesoraphidia heteroneura Ren, 1997, stat. rest. The singularity of this rich paleodiversity could be due to the paleogeographic isolation of the Iberian territory and also the prevalence of wildfires during the Cretaceous.     

The Larval Eye of Nannochoristid Scorpionflies (Insecta,Mecoptera)

Abstract The stemmata of last–instar Nannochoristalarvae are compound eyes composed of 10 or more ommatidia. Each ommatidium has four Semper cells, four distal and four proximal retinula cells which form a cruciform and layered rhabdom. The ommatidia are separated by epidermal cells (possibly rudimentary pigment cells). Corneal lenses are lacking. At the posterior edge, aberrant stemma units may be present which lack a dioptric apparatus and have a star–shaped rhabdom composed of at least six retinula cells. The stemmata of Nannochoristaappear to be derived from stemmata of the Panorpa-type (Mecoptera-Panorpidae). Differences between the stemmata of Nannochoristaand Panorpacan be explained as adaptations to aquatic life (flat cornea) or as regression. A compound larval eye is ascribed to the ground plan of the Mecoptera sensu latoand is considered a genuine plesiomorphy. The identical basic number (seven) of stemmata in the Neuropteroid/Coleoptera assemblage, Amphiesmenoptera and some Mecoptera (Bittacidae, Boreidae) is attributed to parallel evolution.     

The phylogeny and classification of Embioptera (Insecta)

A phylogenetic analysis of the order Embioptera is presented with a revised classification based on results of the analysis. Eighty‐two species of Embioptera are included from all families except Paedembiidae Ross and Embonychidae Navás. Monophyly of each of the eight remaining currently recognized families is tested except Andesembiidae Ross, for which only a single species was included. Nine outgroup taxa are included from Blattaria, Grylloblattaria, Mantodea, Mantophasmatodea, Orthoptera, Phasmida and Plecoptera. Ninety‐six morphological characters were analysed along with DNA sequence data from the five genes 16S rRNA, 18S rRNA, 28S rRNA, cytochrome c oxidase I and histone III. Data were analysed in combined analyses of all data using parsimony and Bayesian optimality criteria, and combined molecular data were analysed using maximum likelihood. Several major conclusions about Embioptera relationships and classification are based on interpretation of these analyses. Of eight families for which monophyly was tested, four were found to be monophyletic under each optimality criterion: Clothodidae Davis, Anisembiidae Davis, Oligotomidae Enderlein and Teratembiidae Krauss. Australembiidae Ross was not recovered as monophyletic in the likelihood analysis in which one Australembia Ross species was recovered in a position distant from other australembiids. This analysis included only molecular data and the topology was not strongly supported. Given this, and because parsimony and the Bayesian analyses recovered a strongly supported clade including all Australembiidae, we regard this family also as monophyletic. Three other families – Notoligotomidae Davis, Archembiidae Ross and Embiidae Burmeister, as historically delimited – were not found to be monophyletic under any optimality criterion. Notoligotomidae is restricted here to include only the genus Notoligotoma Davis with a new family, Ptilocerembiidae Miller and Edgerly, new family, erected to include the genus Ptilocerembia Friederichs. Archembiidae is restricted here to include only the genera Archembia Ross and Calamoclostes Enderlein. The family group name Scelembiidae Ross is resurrected from synonymy with Archembiidae (new status) to include all other genera recently placed in Archembiidae. Embiidae is not demonstrably monophyletic with species currently placed in the family resolved in three separate clades under each optimality criterion. Because taxon sampling is not extensive within this family in this analysis, no changes are made to Embiidae classification. Relationships between families delimited herein are not strongly supported under any optimality criterion with a few exceptions. Either Clothodidae Davis (parsimony) or Australembiidae Ross (Bayesian) is the sister to the remaining Embioptera taxa. The Bayesian analysis includes Australembiidae as the sister to all other Embioptera except Clothididae, suggesting that each of these taxa is a relatively plesiomorphic representatative of the order. Oligotomidae and Teratembiidae are sister groups, and Archembiidae (sensu novum), Ptilocerembiidae, Andesembiidae and Anisembiidae form a monophyletic group under each optimality criterion. Each family is discussed in reference to this analysis, diagnostic combinations and taxon compositions are provided, and a key to families of Embioptera is included.     

Heptageniidae (Insecta,Ephemeroptera) of Thailand

Nine genera and twenty-two species of heptageniid mayflies from Thailand are defined in this present work as well as one suggested further subgenus, Compsoneuria (Siamoneuria) kovaci (species “incertae sedis”) including some particular characters. Taxonomic remarks, diagnoses, line drawings of key characters, distribution, habitat and biological data, and a larval key to the genera and species are provided. The chorionic eggs of eight genera and eight species were observed and shown using a scanning electron microscope.