ABSTRACT
Sensory and motor systems in insects with hemimetabolous development must be ready to mediate adaptive behavior directly on hatching from the egg. For the desert locust S. gregaria, cholinergic transmission from antennal sensillae to olfactory or mechanosensory centers in the brain requires that choline acetyltransferase (ChAT) and the vesicular acetylcholine transporter (vAChT) already be present in sensory cells in the first instar. In this study, we used immunolabeling to demonstrate that ChAT and vAChT are both expressed in sensory cells from identifiable sensilla types in the immature antennal nervous system. We observed ChAT expression in dendrites, neurites and somata of putative basiconic-type sensillae at the first instar stage. We also detected vAChT in the sensory axons of these sensillae in a major antennal nerve tract. We then examined whether evidence for cholinergic transmission is present during embryogenesis. Immunolabeling confirms that vAChT is expressed in somata typical of campaniform sensillae, as well as in small sensory cell clusters typically associated with either a large basiconic or coeloconic sensilla, at 99% of embryogenesis. The vAChT is also expressed in the somata of these sensilla types in multiple antennal regions at 90% of embryogenesis, but not at earlier (70%) embryonic stages. Neuromodulators are known to appear late in embryogenesis in neurons of the locust central complex, and the cholinergic system of the antenna may also only reach maturity shortly before hatching.
Subject(s)
Choline O-Acetyltransferase/metabolism , Grasshoppers/cytology , Nervous System/cytology , Sensilla/cytology , Vesicular Acetylcholine Transport Proteins/metabolism , AnimalsABSTRACT
Juvenile hormone (JH) is known to promote cell polyploidization for insect vitellogenesis and egg production, but the underlying mechanisms remain poorly understood. Using the migratory locust Locusta migratoria as a model system, we report here that the expression of cyclin-dependent kinase 6 (Cdk6) and adenovirus E2 factor-1 (E2f1), the core mediators in cell cycle progression is regulated by JH and its receptor Methoprene-tolerant (Met). JH acts through its receptor complex comprised of Met and Taiman to directly activate the transcription of Cdk6 and E2f1. Depletion of Cdk6 or E2f1 results in significantly decreased ploidy, precocious mitotic entry and increased cell numbers in the fat body, accompanied by substantial reduction of Vitellogenin gene expression, blocked ovarian growth and arrested oocyte maturation. These findings indicate a crucial role of Cdk6 and E2f1 in JH-regulated polyploidization and vitellogenesis as well as a novel regulatory machinery for endocycling in insects.
Subject(s)
Cyclin-Dependent Kinase 6/blood , E2F1 Transcription Factor/biosynthesis , Fat Body/metabolism , Grasshoppers/metabolism , Insect Proteins/biosynthesis , Juvenile Hormones/metabolism , Polyploidy , Transcription, Genetic/physiology , Vitellogenesis/physiology , Animals , Fat Body/cytology , Female , Grasshoppers/cytologyABSTRACT
To understand the role of hyperthermia (HT) in adaptive response, methyl methanesulfonate (MMS) adapted meiotic cells of Poecilocerus pictus were used. Poecilocerus pictus were treated with conditioning (L) or challenging (H) dose of MMS and 2-h time lag (TL) between these doses (L-2h-H) (combined) was employed. Different treatment schedules were used to analyse the influence of HT on MMS-induced adaptive response namely pre; inter; post-treatment and cross-adaptation. After each treatment schedules, chromosomal anomalies were analysed. The frequencies of chromosomal anomalies induced by conditioning and challenging doses of MMS were significantly higher (P < 0.0001) compared to that of the control or HT groups. The combined treatments resulted in significant reduction of chromosomal anomalies compared to additive effect of MMS (P < 0.0001). The pre, inter, post and cross-adaptation treatments with HT reduced the frequencies of chromosomal anomalies compared to the challenge and combined treatments with MMS. There is a protection against MMS-induced chromosomal anomalies by HT in in vivo P.pictus. This is the first report to demonstrate that HT enhances the MMS-induced adaptive response in in vivo meiotic cells.
Subject(s)
Adaptation, Physiological/genetics , DNA Repair/drug effects , Methyl Methanesulfonate/toxicity , Mutagens/toxicity , Adaptation, Physiological/drug effects , Animals , Bone Marrow Cells/drug effects , Chromosome Aberrations/drug effects , DNA Repair/genetics , Grasshoppers/cytology , Meiosis/drug effectsABSTRACT
The tritocerebral commissure giant (TCG) of the grasshopper Schistocerca gregaria is one of the best anatomically and physiologically described arthropod brain neurons. A member of the so-called Ventral Giant cluster of cells, it integrates sensory information from visual, antennal and hair receptors, and synapses with thoracic motor neurons in order to initiate and regulate flight behavior. Its ontogeny, however, remains unclear. In this study, we use bromodeoxyuridine incorporation and cyclin labeling to reveal proliferative neuroblasts in the region of the embryonic brain where the ventral giant cluster is located. Engrailed labeling confirms the deutocerebral identity of this cluster. Comparison of soma locations and initial neurite projections into tracts of the striate deutocerebrum help identify the cells of the ventral cluster in both the embryonic and adult brain. Reconstructions of embryonic cell lineages suggest deutocerebral NB1 as being the putative neuroblast of origin. Intracellular dye injection coupled with immunolabeling against neuron-specific horseradish peroxidase is used to identify the VG1 (TCG) and VG3 neurons from the ventral cluster in embryonic brain slices. Dye injection and backfilling are used to document axogenesis and the progressive expansion of the dendritic arbor of the TCG from mid-embryogenesis up to hatching. Comparative maps of embryonic neuroblasts from several orthopteroid insects suggest equivalent deutocerebral neuroblasts from which the homologous TCG neurons already identified in the adult brain could originate. Our data offer the prospect of identifying further lineage-related neurons from the cluster and so understand a brain connectome from both a developmental and evolutionary perspective.
Subject(s)
Embryo, Nonmammalian/cytology , Grasshoppers/cytology , Grasshoppers/growth & development , Neurons/cytology , Animals , Brain/growth & development , Brain/physiology , Embryo, Nonmammalian/physiology , Grasshoppers/physiology , Neurons/physiologyABSTRACT
We have investigated the pattern of apoptosis in the antennal epithelium during embryonic development of the grasshopper Schistocerca gregaria. The molecular labels lachesin and annulin reveal that the antennal epithelium becomes subdivided into segment-like meristal annuli within which sensory cell clusters later differentiate. To determine whether apoptosis is involved in the development of such sensory cell clusters, we examined the expression pattern of the cell death labels acridine orange and TUNEL in the epithelium. We found stereotypic, age-dependent, wave-like patterns of cell death in the antenna. Early in embryogenesis, apoptosis is restricted to the most basal meristal annuli but subsequently spreads to encompass almost the entire antenna. Cell death then declines in more basal annuli and is only found in the tip region later in embryogenesis. Apoptosis is restricted throughout to the midregion of a given annulus and away from its border with neighboring annuli, arguing against a causal role in annular formation. Double-labeling for cell death and sensory cell differentiation reveals apoptosis occurring within bands of differentiating sensory cell clusters, matching the meristal organization of the apical antenna. Examination of the individual epithelial lineages which generate sensory cells reveals that apoptosis begins peripherally within a lineage and with age expands to encompass the differentiated sensory cell at the base. We conclude that complete lineages can undergo apoptosis and that the youngest cells in these lineages appear to die first, with the sensory neuron dying last. Lineage-based death in combination with cell death patterns in different regions of the antenna may contribute to odor-mediated behaviors in the grasshopper.
Subject(s)
Arthropod Antennae/physiology , Grasshoppers/cytology , Grasshoppers/physiology , Animals , Apoptosis , Arthropod Antennae/embryology , Cell Differentiation , Cell Lineage , Cells, Cultured , Embryonic Development , Epithelial Cells/cytology , Grasshoppers/embryology , Neurons/cytologyABSTRACT
A detailed account is given by the octopaminergic innervation of the antennal heart in Schistocerca gregaria using various immunohistochemical methods. Anterograde axonal filling illustrates the unilateral innervation on the medial ventral surface of the pumping muscle of the antennal heart via the paired corpora cardiaca nerve III. In addition, antibody staining revealed that ascending axons of this nerve terminate at the ampullae of the antennal heart forming synaptoid structures and extensive neurohaemal release sites. Due to the innervation by two dorsal unpaired median neurons, the presence of the biogenic amines octopamine and tyramine could be visualized by immunocytochemistry in an insect antennal heart for the first time. The data suggest that tyramine acts as a precursor and not purely as an independent transmitter. While the octopaminergic fibers innervating the pumping muscle of the antennal heart indicate a cardioregulatory role, we conclude that octopamine released from the neurohaemal area is pumped into the antennae and an involvement in the modulation of the antennal sensory sensitivity is discussed.
Subject(s)
Arthropod Antennae/innervation , Arthropod Antennae/metabolism , Grasshoppers/cytology , Grasshoppers/metabolism , Octopamine/metabolism , Animals , Biotin/analogs & derivatives , Biotin/metabolism , Cardiovascular System/anatomy & histology , Cardiovascular System/metabolism , Female , Insect Proteins/metabolism , Male , Muscles/innervation , Muscles/metabolism , Neurons/cytology , Neurons/metabolism , Streptavidin/metabolism , Synapsins/metabolism , Tyramine/metabolismABSTRACT
The central complex comprises an elaborate system of modular neuropils which mediate spatial orientation and sensory-motor integration in insects such as the grasshopper and Drosophila. The neuroarchitecture of the largest of these modules, the fan-shaped body, is characterized by its stereotypic set of decussating fiber bundles. These are generated during development by axons from four homologous protocerebral lineages which enter the commissural system and subsequently decussate at stereotypic locations across the brain midline. Since the commissural organization prior to fan-shaped body formation has not been previously analyzed in either species, it was not clear how the decussating bundles relate to individual lineages, or if the projection pattern is conserved across species. In this study, we trace the axonal projections from the homologous central complex lineages into the commissural system of the embryonic and larval brains of both the grasshopper and Drosophila. Projections into the primordial commissures of both species are found to be lineage-specific and allow putatively equivalent fascicles to be identified. Comparison of the projection pattern before and after the commencement of axon decussation in both species reveals that equivalent commissural fascicles are involved in generating the columnar neuroarchitecture of the fan-shaped body. Further, the tract-specific columns in both the grasshopper and Drosophila can be shown to contain axons from identical combinations of central complex lineages, suggesting that this columnar neuroarchitecture is also conserved.
Subject(s)
Drosophila/growth & development , Grasshoppers/growth & development , Animals , Axons/metabolism , Biological Evolution , Body Patterning , Brain/cytology , Drosophila/cytology , Drosophila/genetics , Drosophila/metabolism , Grasshoppers/cytology , Grasshoppers/genetics , Grasshoppers/metabolism , Larva/growth & development , Larva/metabolism , Neurons , Neuropil/cytologyABSTRACT
The nervous system of the antenna of the grasshopper Schistocerca gregaria consists of two nerve tracts in which sensory cells project their axons to the brain. Each tract is pioneered early in embryogenesis by a pair of identified cells located apically in the antennal lumen. The pioneers are thought to originate in the epithelium of the antenna and then delaminate into the lumen where they commence axogenesis. However, unambiguous molecular identification of these cells in the epithelium, of an identifiable precursor, and of their mode of generation has been lacking. In this study, we have used immunolabeling against neuron-specific horseradish peroxidase and against Lachesin, a marker for differentiating epithelial cells, in combination with the nuclear stain DAPI, to identify the pioneers within the epithelium of the early embryonic antenna. We then track their delamination into the lumen as differentiated neurons. The pioneers are not labeled by the mesodermal/mesectodermal marker Mes3, consistent with an epithelial (ectodermal) origin. Intracellular dye injection, as well as labeling against the mitosis marker phospho-histone 3, identifies precursor cells in the epithelium, each associated with a column of cells. Culturing with the S-phase label 5-ethynyl-2'-deoxyuridine (EdU) shows that both a precursor and its column have incorporated the label, confirming a lineage relationship. Each set of pioneers can be shown to belong to a separate lineage of such epithelial cells, and the precursors remain and are proliferative after generating the pioneers. Analyses of mitotic spindle orientation then enable us to propose a model in which a precursor generates its pioneers asymmetrically via self-renewal.
Subject(s)
Grasshoppers/growth & development , Animals , Arthropod Antennae/growth & development , Axons/physiology , Epithelial Cells/cytology , Epithelial Cells/physiology , Grasshoppers/cytology , Grasshoppers/physiology , Mitosis , Nervous System/cytology , Nervous System/growth & development , Neurons/cytology , Spindle Apparatus , Stem Cells/cytology , Stem Cells/physiologyABSTRACT
Desert locusts (Schistocerca gregaria) sense the plane of dorsally presented polarized light through specialized dorsal eye regions that are likely adapted to exploit the polarization pattern of the blue sky for spatial orientation. Receptive fields of these dorsal rim photoreceptors and polarization-sensitive interneurons are directed toward the upper sky but may extend to elevations below 30°. Behavioral data, however, suggests that S. gregaria is even able to detect polarized light from ventral directions but physiological evidence for this is still lacking. In this study we characterized neurons in the locust brain showing polarization sensitivity at low elevations down to the horizon. In most neurons polarization sensitivity was absent or weak when stimulating from the zenith. All neurons, including projection and commissural neurons of the optic lobe and local interneurons of the central brain, are novel cell types, distinct from polarization-sensitive neurons studied so far. Painting dorsal rim areas in both eyes black to block visual input had no effect on the polarization sensitivity of these neurons, suggesting that they receive polarized light input from the main eye. A possible role of these neurons in flight stabilization or the perception of polarized light reflected from bodies of water or vegetation is discussed.
Subject(s)
Grasshoppers/physiology , Grasshoppers/radiation effects , Light , Neurons/physiology , Neurons/radiation effects , Sensation/physiology , Action Potentials , Animals , Brain/cytology , Brain/physiology , Brain/radiation effects , Grasshoppers/cytology , Microelectrodes , Microscopy, Confocal , Microscopy, Fluorescence , Neurons/cytology , Photic StimulationABSTRACT
The central complex of the insect brain comprises a group of neuropils involved in spatial orientation and memory. In fruit flies it mediates place learning based on visual landmarks and houses neurons that encode the orientation for goal-directed locomotion, based on landmarks and self-motion cues for angular path-integration. In desert locusts, the central complex holds a compass-like representation of head directions, based on the polarization pattern of skylight. Through intracellular recordings from immobilized locusts, we investigated whether sky compass neurons of the central complex also represent the position or any salient feature of possible landmarks, in analogy to the observations in flies. Neurons showed strongest responses to the novel appearance of a small moving square, but we found no evidence for a topographic representation of object positions. Responses to an individual square were independent of direction of motion and trajectory, but showed rapid adaptation to successive stimulation, unaffected by changing the direction of motion. Responses reappeared, however, if the moving object changed its trajectory or if it suddenly reversed moving direction against the movement of similar objects that make up a coherent background-flow as induced by ego-motion. Response amplitudes co-varied with the precedent state of dynamic background activity, a phenomenon that has been related to attention-dependent saliency coding in neurons of the mammalian primary visual cortex. The data show that neurons of the central complex of the locust brain are visually bimodal, signaling sky compass direction and the novelty character of moving objects. These response properties might serve to attune compass-aided locomotor control to unexpected events in the environment. The difference to data obtained in fruit flies might relate to differences in the lifestyle of landmark learners (fly) and compass navigators (locust), point to the existence of parallel networks for the two orientation strategies, or reflect differences in experimental conditions.
Subject(s)
Brain/cytology , Brain/physiology , Grasshoppers/cytology , Grasshoppers/physiology , Vision, Ocular/physiology , Animals , Drosophila , MotionABSTRACT
The twin nerve tracts of the antenna of the grasshopper Schistocerca gregaria are established early in embryogenesis by sibling pairs of pioneers which delaminate from the epithelium into the lumen at the antennal tip. These cells can be uniquely identified via their co-expression of the neuronal labels horseradish peroxidase and the lipocalin Lazarillo. The apical pioneers direct axons toward the antennal base where they encounter guidepost-like cells called base pioneers which transiently express the same molecular labels as the apical pioneers. To what extent the pioneer growth cones then progress into the brain neuropil proper, and what their targets there might be, has remained unclear. In this study, we show that the apical antennal pioneers project centrally beyond the antennal base first into the deutocerebral, and then into the protocerebral brain neuropils. In the protocerebrum, we identify their target circuitry as being identified Lazarillo-positive cells which themselves pioneer the primary axon scaffold of the brain. The apical and base antennal pioneers therefore form part of a molecularly contiguous pathway from the periphery to an identified central circuit of the embryonic grasshopper brain.
Subject(s)
Arthropod Antennae/embryology , Grasshoppers/embryology , Nervous System/embryology , Neurons/physiology , Animals , Arthropod Antennae/cytology , Axons/physiology , Brain/cytology , Brain/embryology , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/embryology , Grasshoppers/cytology , Models, Neurological , Nervous System/cytology , Neuropil/cytology , Neuropil/physiologyABSTRACT
Small-diameter nerves were found that are associated with the lateral peripheral nerves of the unfused abdominal ganglia of locusts. Such small nerves were observed in about 30% of all cases in Locusta migratoria, more than 60% in Schistocerca gregaria. Retrograde staining of these small nerves showed two somata in the posterior, lateral, and ventral region of an abdominal ganglion. These cells give rise to the small nerves that accompany the big lateral nerves and, on their surface, form putative neurohaemal release sites. Astonishingly the cells do not form any dendritic ramifications within the neuropile of the ganglia.
Subject(s)
Grasshoppers/cytology , Animals , Dendrites , Female , Locusta migratoria/cytology , Male , Microscopy, Confocal , Neurons/cytology , Neurosecretory Systems/cytologyABSTRACT
The small size of some insects, and the crystalline regularity of their eyes, have made them ideal for large-scale reconstructions of visual circuits. In phylogenetically recent muscomorph flies, like Drosophila, precisely coordinated output to different motion-processing pathways is delivered by photoreceptors (R cells), targeting four different postsynaptic cells at each synapse (tetrad). Tetrads were linked to the evolution of aerial agility. To reconstruct circuits for vision in the larger brain of a locust, a phylogenetically old, flying insect, we adapted serial block-face scanning electron microscopy (SBEM). Locust lamina monopolar cells, L1 and L2, were the main targets of the R cell pathway, L1 and L2 each fed a different circuit, only L1 providing feedback onto R cells. Unexpectedly, 40% of all locust R cell synapses onto both L1 and L2 were tetrads, revealing the emergence of tetrads in an arthropod group present 200 million years before muscomorph flies appeared, coinciding with the early evolution of flight.
Subject(s)
Biological Evolution , Grasshoppers/cytology , Photoreceptor Cells, Invertebrate/cytology , Synapses/ultrastructure , Visual Pathways/cytology , Animals , Brain/cytology , Brain/metabolism , Grasshoppers/metabolism , Imaging, Three-Dimensional , Insect Proteins/metabolism , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Photoreceptor Cells, Invertebrate/metabolism , Synapses/metabolism , Taurine/metabolism , Visual Pathways/metabolismABSTRACT
The antennal nervous system of the grasshopper Schistocerca gregaria comprises two parallel pathways projecting to the brain, each pioneered early in embryogenesis by a pair of sibling cells located at the antennal tip. En route, the growth cones of pioneers from one pathway have been shown to contact a guidepost-like cell called the base pioneer. Its role in axon guidance remains unclear as do the cellular guidance cues regulating axogenesis in the other pathway supposedly without a base pioneer. Further, while the tip pioneers are known to delaminate from the antennal epithelium into the lumen, the origin of this base pioneer is unknown. Here, we use immunolabeling and immunoblocking methods to clarify these issues. Co-labeling against the neuron-specific marker horseradish peroxidase and the pioneer-specific cell surface glycoprotein Lazarillo identifies not only the tip pioneers but also a base pioneer associated with each of the developing antennal pathways. Both base pioneers co-express the mesodermal label Mes3, consistent with a lumenal origin, whereas the tip pioneers proved Mes3-negative confirming their affiliation with the ectodermal epithelium. Lazarillo antigen expression in the antennal pioneers followed a different temporal dynamic: continuous in the tip pioneers, but in the base pioneers, only at the time their filopodia and those of the tip pioneers first recognize one another. Immunoblocking of Lazarillo expression in cultured embryos disrupts this recognition resulting in misguided axogenesis in both antennal pathways.
Subject(s)
Arthropod Antennae/growth & development , Grasshoppers/growth & development , Animals , Axons/metabolism , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/metabolism , Grasshoppers/cytology , Grasshoppers/metabolism , Nervous System/cytology , Nervous System/growth & developmentABSTRACT
The single-copy gene Period was located in five grasshopper species belonging to the Acridomorpha group through permanent in situ hybridization (PISH). The mapping revealed one copy of this gene in the L1 chromosome pair in Ommexecha virens, Xyleus discoideus angulatus, Tropidacris collaris, Schistocerca pallens, and Stiphra robusta. A possible second copy was mapped on the L2 chromosome pair in S. robusta, which should be confirmed by further studies. Except for the latter case, the chromosomal position of the Period gene was highly conserved among the four families studied. The S. robusta karyotype also differs from the others both in chromosome number and morphology. The position conservation of the single-copy gene Period contrasts with the location diversification of multigene families in these species. The localization of single-copy genes by PISH can provide new insights about the genomic content and chromosomal evolution of grasshoppers and others insects.
Subject(s)
Grasshoppers/classification , Grasshoppers/genetics , In Situ Hybridization, Fluorescence/methods , Period Circadian Proteins/genetics , Physical Chromosome Mapping/methods , Animals , Chromosomes, Insect , Conserved Sequence , Evolution, Molecular , Gene Dosage , Grasshoppers/cytology , Karyotype , MeiosisABSTRACT
In vitro primary cultures of dissociated invertebrate neurons from locust ganglia are used to experimentally investigate the morphological evolution of assemblies of living neurons, as they self-organize from collections of separated cells into elaborated, clustered, networks. At all the different stages of the culture's development, identification of neurons' and neurites' location by means of a dedicated software allows to ultimately extract an adjacency matrix from each image of the culture. In turn, a systematic statistical analysis of a group of topological observables grants us the possibility of quantifying and tracking the progression of the main network's characteristics during the self-organization process of the culture. Our results point to the existence of a particular state corresponding to a small-world network configuration, in which several relevant graph's micro- and meso-scale properties emerge. Finally, we identify the main physical processes ruling the culture's morphological transformations, and embed them into a simplified growth model qualitatively reproducing the overall set of experimental observations.
Subject(s)
Grasshoppers/cytology , Neurons/cytology , Animals , Cells, Cultured , Models, Neurological , Nerve Net/cytology , Nerve Net/physiologyABSTRACT
This study employs labels for cell proliferation and cell death, as well as classical histology to examine the fates of all eight neural stem cells (neuroblasts) whose progeny generate the central complex of the grasshopper brain during embryogenesis. These neuroblasts delaminate from the neuroectoderm between 25 and 30 % of embryogenesis and form a linear array running from ventral (neuroblasts Z, Y, X, and W) to dorsal (neuroblasts 1-2, 1-3, 1-4, and 1-5) along the medial border of each protocerebral hemisphere. Their stereotypic location within the array, characteristic size, and nuclear morphologies, identify these neuroblasts up to about 70 % of embryogenesis after which cell shrinkage and shape changes render progressively more cells histologically unrecognizable. Molecular labels show all neuroblasts in the array are proliferative up to 70 % of embryogenesis, but subsequently first the more ventral cells (72-75 %), and then the dorsal ones (77-80 %), cease proliferation. By contrast, neuroblasts elsewhere in the brain and optic lobe remain proliferative. Apoptosis markers label the more ventral neuroblasts first (70-72 %), then the dorsal cells (77 %), and the absence of any labeling thereafter confirms that central complex neuroblasts have exited the cell cycle via programmed cell death. Our data reveal appearance, proliferation, and cell death proceeding as successive waves from ventral to dorsal along the array of neuroblasts. The resulting timelines offer a temporal blueprint for building the neuroarchitecture of the various modules of the central complex.
Subject(s)
Grasshoppers/cytology , Grasshoppers/growth & development , Animals , Apoptosis , Brain/cytology , Brain/metabolism , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/metabolism , Grasshoppers/metabolism , Mitosis , Neural Stem Cells/cytology , Neural Stem Cells/metabolismABSTRACT
It is well established that the brains of adult malacostracan crustaceans and winged insects display distinct homologies down to the level of single neuropils such as the central complex and the optic neuropils. We wanted to know if developing insect and crustacean brains also share similarities and therefore have explored how neurotransmitter systems arise during arthropod embryogenesis. Previously, Sintoni et al. (2007) had already reported a homology of an individually identified cluster of neurons in the embryonic crayfish and insect brain, the secondary head spot cells that express the Engrailed protein. In the present study, we have documented the ontogeny of the serotonergic system in embryonic brains of the Marbled Crayfish in comparison to Migratory Locust embryos using immunohistochemical methods combined with confocal laser-scan microscopy. In both species, we found a cluster of early emerging serotonin-immunoreactive neurons in the protocerebrum with neurites that cross to the contralateral brain hemisphere in a characteristic commissure suggesting a homology of this cell cluster. Our study is a first step towards a phylogenetic analysis of neurotransmitter system development and shows that, as for the ventral nerve cord, traits related to neurogenesis in the brain can provide valuable hints for resolving the much debated question of arthropod phylogeny.
Subject(s)
Astacoidea/embryology , Brain/embryology , Grasshoppers/embryology , Nerve Net/cytology , Animal Migration , Animals , Astacoidea/cytology , Astacoidea/growth & development , Brain/cytology , Grasshoppers/cytology , Grasshoppers/growth & development , Immunohistochemistry , Nerve Net/drug effects , Nerve Net/ultrastructure , Neurotransmitter Agents , Serotonin/metabolism , Serotonin/pharmacologyABSTRACT
I investigate the pattern of gliogenesis in the brain of the grasshopper Schistocerca gregaria prior to mid-embryogenesis, with particular focus on the protocerebrum. Using the glia-specific marker Repo and the neuron-specific marker HRP, I identify three types of glia with respect to their respective positions in the brain: surface glia form the outmost cell layer ensheathing the brain; cortex glia are intermingled with neuronal somata forming the brain cortex; and neuropil glia are associated with brain neuropils. The ontogeny of each glial type has also been studied. At 24% of embryogenesis, a few glia are observed in each hemisphere of the proto-, deuto- and tritocerebrum. In each protocerebral hemisphere, such glia form a cluster that expands rapidly during later development. Closer examination reveals proliferative glia in such clusters at ages spanning from 24 to 36% of embryogenesis, indicating that glial proliferation may account for the expansion of the clusters. Data derived from 33-39% of embryogenesis suggest that, in the protocerebrum, each type of glia is likely to be generated by its respective progenitor-forming clusters. Moreover, the glial cluster located at the anterior end of the brain can give rise to both surface glia and cortex glia that populate the protocerebrum via subsequent migration. Proliferation is observed for all three glial types, indicating a possible source for the glia.
Subject(s)
Grasshoppers/embryology , Animals , Brain/cytology , Brain/embryology , Brain/growth & development , Cell Growth Processes/physiology , Embryonic Development , Grasshoppers/cytology , Grasshoppers/metabolism , Immunohistochemistry , Neuroglia/cytology , Neuroglia/metabolism , Neuroglia/physiologyABSTRACT
Supernumerary chromosomes (B chromosomes) occur in approximately 15% of eukaryote species. Although these chromosomes have been extensively studied, knowledge concerning their specific molecular composition is lacking in most cases. The accumulation of repetitive DNAs is one remarkable characteristic of B chromosomes, and the occurrence of distinct types of multigene families, satellite DNAs and some transposable elements have been reported. Here, we describe the organization of repetitive DNAs in the A complement and B chromosome system in the grasshopper species Abracris flavolineata using classical cytogenetic techniques and FISH analysis using probes for five multigene families, telomeric repeats and repetitive C0t-1 DNA fractions. The 18S rRNA and H3 histone multigene families are highly variable and well distributed in A. flavolineata chromosomes, which contrasts with the conservation of U snRNA genes and less variable distribution of 5S rDNA sequences. The H3 histone gene was an extensively distributed with clusters occurring in all chromosomes. Repetitive DNAs were concentrated in C-positive regions, including the pericentromeric region and small chromosomal arms, with some occurrence in C-negative regions, but abundance was low in the B chromosome. Finally, the first demonstration of the U2 snRNA gene in B chromosomes in A. flavolineata may shed light on its possible origin. These results provide new information regarding chromosomal variability for repetitive DNAs in grasshoppers and the specific molecular composition of B chromosomes.