Simulation of laser-induced ionization in wide bandgap solid dielectrics with a particle-in-cell code.

Modeling the laser-plasma interaction within solids is crucial in controlling ultrafast laser processing of dielectrics, where the pulse propagation and plasma formation dynamics are highly intricate. This is especially important when dealing with nano-scale plasmas where specific phenomena of plasma physics, such as resonance absorption, can significantly impact the energy deposition process. In this article, we report on adapting of a Particle-In-Cell code, EPOCH, to model the laser-plasma interaction within solids. This is performed by implementing a background permittivity and by developing and validating adapted field ionization and impact ionization modules. They are based on the Keldysh ionization theory and enable the modeling of ionization processes within solids. The implementation of these modules was validated through comparisons with a hydrodynamic code and existing literature. We investigate the necessary number of super-particles per cell to model realistic ionization dynamics. Finally, we apply the code to explore the dynamics of plasma formation in the regime of of quantized structuring of transparent films. Our study elucidates how a stack of nano-plasma layers can be formed by the interference of a pulse with its reflection on the exit surface of a high refractive index material.

Short- and long-chain natural polyamines play specific roles in adult cricket neuroblast proliferation and neuron differentiation in vitro.

In the house cricket (Acheta domesticus) mushroom bodies, neurogenesis still occurs during adulthood. Using in vitro approaches, the respective roles of natural polyamines in neurogenesis were examined. Mushroom body neuroblast proliferation was assayed in organotypic culture using 5-bromo, 2'-deoxyuridine labeling. The number of labeled cells was significantly increased when putrescine was added to culture medium, whereas spermidine and spermine supplementation did not alter cell proliferation. Conversely, in vitro morphometric studies on mushroom body neurons cultured in a defined medium showed that putrescine addition failed to alter any morphological character of these interneurons, whereas addition of the long-chain polyamines, spermidine and spermine, stimulated neuron differentiation. These two polyamines significantly increased total neurite length; moreover, spermidine-treated cells exhibited more branches than the controls. The present data demonstrate that putrescine has a mitogenic effect on mushroom body neuronal precursors, and that spermidine and spermine, which failed to induce neuroblast proliferation, act on neuronal differentiation, inducing neurite outgrowth. Our results indicate that short- and long-chain polyamines play specific roles during neurogenesis, and provide a basis for further studies on neuronal precursor proliferation and differentiation.

Influence of environmental stimulation on neurogenesis in the adult insect brain.

Mushroom bodies are the main integrative structures of insect brain. They receive sensory information from the eyes, the palps, and the antennae. In the house cricket, Acheta domesticus, a cluster of mushroom body neuroblasts keeps producing new interneurons during an insect's life span. The aim of the present work is to study the impact of environmental stimuli on mushroom body neurogenesis during adulthood. Crickets were reared either in an enriched environment, where they received complex environmental and congeneric stimulations or isolated in small cages and deprived of most visual, auditory, and olfactory stimuli. They then were injected with a S-phase marker, 5-bromo, 2'-deoxyuridine (BrdU) and sacrificed at different periods of their life. Neurogenesis and cell survival were estimated by counting the number of BrdU-labeled cells in the mushroom bodies. Environmentally enriched crickets were found to have an increased number of newborn cells in their mushroom bodies compared with crickets housed in cages with an impoverished environment. This effect of external factors on neurogenesis seems to be limited to the beginning of imaginal life. Furthermore, no cell loss could be detected among the newborn neurons in either environmental situation, suggesting that cell survival was not affected by the quality of the environment. Considering vertebrate studies which showed that enriched environment increases hippocampal cell survival and improves animal performances in spatial learning tests, we suggest that the increased number of interneurons produced in an integrative brain structure after exposure to enriched environment could contribute to adaptive behavioral performances in adult insects.

Dual effect of ecdysone on adult cricket mushroom bodies.

Mushroom bodies, which are the main integrative centre for insect sensorial information, play a critical role in associative olfactory learning and memory. This paired brain structure contains interneurons grouped in a cortex, sending their axons into organized neuropiles. In the house cricket (Acheta domesticus) brain, persistent neuroblasts proliferate throughout adult life. Juvenile hormone (JH) has been shown to stimulate this proliferation [Cayre, M., Strambi, C. & Strambi, A. (1994) Nature, 368, 57-59]. In the present study, the effect of morphogenetic hormones on mushroom body cells maintained in primary culture was examined. Whereas JH did not significantly affect neurite growth, ecdysone significantly stimulated neurite elongation. Moreover, ecdysone also acted on neuroblast proliferation, as demonstrated by the reduced number of cells labelled with 5-bromodeoxyuridine following ecdysone application. Heterospecific antibodies raised against ecdysone receptor protein and ultraspiracle protein, the two heterodimers of ecdysteroid receptors, showed positive immunoreactivity in nervous tissue extracts and in nuclei of mushroom body cells, indicating the occurrence of putative ecdysteroid receptors in cricket mushroom body cells. These data indicate a dual role for ecdysone in adult cricket mushroom bodies: this hormone inhibits neuroblast proliferation and stimulates interneuron differentiation. These results suggest that a constant remodelling of mushroom body structure could result from physiological changes in hormone titres during adult life.

c-Fos-related antigens in the central nervous system of an insect, Acheta domesticus.

Fos-related antigens (Fra) were detected in the nuclei of neurones in young adult Acheta domesticus female crickets by immunohistochemical analysis, using an antibody that recognizes the amino-acid sequence 127-152 of c-Fos protein. Specificity of Fra immunoreactivity was confirmed by Western blot analysis of nuclear extracts from neural tissues. A major immunoreactive doublet with an apparent molecular mass of 52,000/54,000 Da was detected in nuclear extracts. Immunostaining of the 52,000/54,000 Da doublet showed variations in intensity during the first 5 days following the imaginal molt. Staining was more intense between day 2 and day 4 when ecdysteroid titers were high. Expression of Fra was low in allatectomized (i.e., deprived of juvenile hormone and ecdysteroids) and ovariectomized (i.e., deprived of ecdysteroids) females as compared to control females. These results show the involvement of hormone-regulated process in expression of Fra. The effect of nociceptive stimulation on Fra expression was tested. Twenty minutes after removal of the ovipositor, a supplementary band with an apparent molecular mass of 70,000 Da appeared in the nuclear extracts, then decreased and disappeared totally after 45 min. Several other Fos-related antigens with different temporal patterns of expression were also detected.

Fate of neuroblast progeny during postembryonic development of mushroom bodies in the house cricket, Acheta domesticus.

Mushroom bodies represent the main sensory integrative center of the insect brain and probably play a major role in the adaptation of behavioral responses to the environment. Taking into account the continuous neurogenesis of cricket mushroom bodies, we investigated ontogenesis of this brain structure. Using BrdU labeling, we examined the fate of neuroblast progeny during the postembryonic development. Preimaginal Kenyon cells survived throughout larval and imaginal moults and persisted during adulthood. Our results indicate that the location of labelled Kenyon cells in the cortex of the adult cricket mainly depends upon the period when they were produced during development. The present data demonstrate that cricket mushroom bodies grow from the inside out and that, at any developmental stage, the center of the cortex contains the youngest Kenyon cells. This study also allowed us to observe the occurrence of quiescent neuroblasts. Kenyon cell death during postembryonic and adult life seems to be reduced. Although preimaginal Kenyon cells largely contribute to adult mushroom body structure, a permanent remodeling of the mushroom body occurs throughout the whole insect life due to the persistence of neurogenesis in the house cricket. Further studies are needed to understand the functional significance of these findings.

Immunocytochemical mapping of an RDL-like GABA receptor subunit and of GABA in brain structures related to learning and memory in the cricket Acheta domesticus.

The distribution of putative RDL-like GABA receptors and of gamma-aminobutyric acid (GABA) in the brain of the adult house cricket Acheta domesticus was studied using specific antisera. Special attention was given to brain structures known to be related to learning and memory. The main immunostaining for the RDL-like GABA receptor was observed in mushroom bodies, in particular the upper part of mushroom body peduncle and the two arms of the posterior calyx. Weaker immunostaining was detected in the distal part of the peduncle and in the alpha and beta lobes. The dorso- and ventrolateral protocerebrum neuropils appeared rich in RDL-like GABA receptors. Staining was also detected in the glomeruli of the antennal lobe, as well as in the ellipsoid body of the central complex. Many neurons clustered in groups exhibit GABA-like immunoreactivity. Tracts that were strongly immunostained innervated both the calyces and the lobes of mushroom bodies. The glomeruli of the antennal lobe, the ellipsoid body, as well as neuropils of the dorso- and ventrolateral protocerebrum were also rich in GABA-like immunoreactivity. The data demonstrated a good correlation between the distribution of the GABA-like and of the RDL-like GABA receptor immunoreactivity. The prominent distribution of RDL-like GABA receptor subunits, in particular areas of mushroom bodies and antennal lobes, underlines the importance of inhibitory signals in information processing in these major integrative centers of the insect brain.

Specific requirement of putrescine for the mitogenic action of juvenile hormone on adult insect neuroblasts.

Persistent neurogenesis in an adult insect brain was recently shown to be stimulated by juvenile hormone (JH). This morphogenetic hormone was also shown to act on polyamine biosynthesis. To analyze the possible involvement of polyamines in the neurogenic action of JH, two series of experiments were carried out with adult female crickets, Acheta domesticus: (i) inhibition of the first key enzyme in polyamine biosynthesis, ornithine decarboxylase, with alpha-difluoromethylornithine (alpha-DFMO), and examination of the effects of this treatment on the neuroblast proliferation response to JH; and (ii) examination of the effects of putrescine supplementation on the mitotic index of JH-deprived and alpha-DFMO-treated females. In control females, alpha-DFMO treatment, as well as JH deprivation, greatly reduced neuroblast proliferation. Putrescine supplementation in alpha-DFMO-treated insects overcame the effects of alpha-DFMO, and allowed for detection of putrescine in the neural tissue and stimulation of brain neurogenesis. In JH-deprived females, alpha-DFMO treatment completely prevented the stimulatory action of JH on neuroblast proliferation and on brain putrescine levels. By contrast, putrescine feeding of JH-deprived animals was able to mimic the stimulatory effect of JH: brain putrescine levels increased and neuroblast proliferation was restored. To our knowledge, this report demonstrates for the first time that in vivo administration of putrescine can mimic the effects of a morphogenetic hormone on adult neuroblast proliferation, and shows the importance of polyamines, especially putrescine, in the transduction of JH message in neural tissue.

Casein kinase II activity in the brain of an insect, Acheta domesticus: characterization and hormonal regulation.

This study documented casein kinase II (CK II) activity in Acheta domesticus brain using specific antibodies and its regulation by polyamines. In control animals a transient decrease in CK II activity at day 3 after imaginal moult was observed in the brain but not in the fat body. If deprived of ecdysone by ovariectomy a different pattern was observed, with CK II activity being significantly higher on days 3 and 4 after emergence. After ecdysone injection in ovariectomized females, CK II activity decreased to levels similar to those in controls. The implications of ecdysone regulation of brain CK II activity are discussed.

Inhibition of polyamine biosynthesis alters oviposition behavior in female crickets.

The role of polyamines in the expression of cricket oviposition, a juvenile hormone-dependent behavior, was investigated using a specific inhibitor of ornithine decarboxylase, alpha-difluoromethylornithine (alpha-DFMO). The fat body of treated female house crickets (Acheta domesticus) did not show any putrescine and presented reduced levels of spermidine, whereas spermine titres were significantly enhanced. In nervous tissue, alpha-DFMO did not affect spermine titres but induced a severe drop in spermidine levels. In polyamine depleted females, the expression of egg-laying behavior was delayed and was expressed less frequently compared with controls. As drug treatment did not seem to affect juvenile hormone titres, the data suggest that juvenile hormone might act on behavior by way of polyamine metabolism. These results support the view that, in insects, as in vertebrates, the ornithine decarboxylase-polyamine system is involved in the maturation of complex behaviors.

Neurogenesis in adult insect mushroom bodies.

The occurrence of neurogenesis in mushroom bodies of adult insects belonging to several orthopteroid and coleopteran families is described. Using injections of 5-bromo, T2'-deoxyuridine, we showed that neuroblasts, which are progenitors of Kenyon cells during preimaginal instars, continue to divide in adult Acheta domesticus. Their progeny constitute a central column in mushroom body cortices of 3-week-old females. Other Gryllidae, Gryllus bimaculatus and Gryllomorpha dalmatina, show the same pattern of neuroblast activity and migration of their progeny. Immunocytochemical staining of glial cells failed to reveal any immunoreactivity, either in proliferating regions or in the resulting cells. In another orthopteran, Locusta migratoria, discrete clusters of cells, located dorsolateral to the Kenyon cells, incorporated 5-bromo, 2'-deoxyuridine, but we could not detect any neuronal progeny migrating to the mushroom body cortices. These cells were strongly labeled with an antiglial antibody, indicating that the replicating cells are glioblasts rather than neuroblasts. In Periplaneta americana (Dictyoptera), cells replicating their DNA were similarly shown to immunoreact with glial antibodies. In contrast, three coleopterans (Tenebrio molitor, Zophobas species, Harmonia axyridis) have two large neuroblasts located in the middle of the mushroom body cortices. These produce cells which migrate within the group of Kenyon cells, their nuclei having the same shape and size as those of surrounding Kenyon cells. In adult insects, neurogenesis in mushroom bodies occurs in Gryllidae and several coleopteran families, but could not be demonstrated in Dictyoptera and Acrididae. Its occurrence and distribution raise the issue of unexpected plasticity in the adult insect brain.

Ovaries and regulation of juvenile hormone titer in Acheta domesticus L. (Orthoptera).

A study was performed on females Acheta domesticus to examine the effects of various experimental conditions on the ovarian physiology. Using a radioimmunoassay to determine juvenile hormone (JH) titers as well as in vitro JH biosynthesis, we observed that retention of mature follicles in egg-retaining females, i.e., virgins or mated females not provided an egg-laying substrate, inhibits JH production and consequently oocyte development. Mating in intact as well as ovariectomized females does not affect corpora allata activity. It is only when mating is associated with egg laying that JH biosynthesis and hemolymph titers increased and oocyte development and fecundity are stimulated. Despite lower JH biosynthesis, ovariectomized females present enlarged corpora allata and the levels of JH observed in their hemolymph were intermediate between those of intact egg-laying and virgin females. In intact females, the hemolymph JH titers as well as the JH esterase activities were related to ovarian development. JH esterase activity was very high in ovariectomized animals. Several factors involved in ovarian development of A. domesticus are discussed.

In vivo application of HRP crystals: a new and efficient method for tracing neural connections in insects.

Solid HRP pellets prepared with a 2.5% Triton X-100 aqueous solution were implanted either into corpora allata or applied onto neurohemal organs of a cricket. The method presents two advantages: it allows one to perform "in vivo" instead of "in vitro" experiments, and detergent HRP pellets are easy to manipulate. Thus, this method combines simplicity with accuracy and appears to be very useful in tracing neural connections in the insect nervous system.

Ultrastructural dynamics of the corpus allatum of Choleva angustata Fab. (Coleoptera, Catopidae).

The ultrastructure of the corpora allata (CA) during postembryonic stages of Choleva angustata Fab. shows cyclic changes, in particular regarding the endoplasmic reticulum and the mitochondria. During the last larval instar, at the short transitory period which follows the cessation of alimentation, we observed a highly unusual appearance of the CA, characterized by intensive vacuolization. After this transitory period, the smooth endoplasmic reticulum (ser) disappears until the imago emerges. These observations along with our experimental data permit a physiological interpretation.

[Ultrastructural development of the corpora allata during the last larval stage of Choleva angustata Fab. (Coleoptera Catopidae of the subfamily Catopinae)]. / Evolution ultrastructurale des corps allates au cours du dernier stade larvaire chez Choleva angustata Fab. (Coléoptéres Catopidae de la sous-famille des Catopinae)

The ultrastructure of the corpora allata of the last larvar instar of Choleva angustata Fab. was studied. In normal development a complex pattern of changes in the ultrastructure of the corpora allata occurs in relation to the larval age in the active and inactive stages. Towards the end of the active stage, a drastic change occurs, involving an increase in number and volume of vacuoles which fill up the cells. During the fasting stage (in the nymphal clay case) the smooth endoplasmic reticulum disappears while the rough endoplasmic reticulum forms long and sinuous cisternae. Towards the end of the instar, numerous vacuoles appear.

[Ultrastructural changes in the corpora allata of the last larval instar of Choleva angustata Fab. (Catopidae). The effect of fasting, with or without ecdysterone]. / Evolution ultrastructurale des corps allates de la larve du dernier stade de Choleva angustata Fab. (Catopidae). Action du jeûne, en présence ou non d'ecdystérone

Fasting experiments show striking changes at the ultrastructural level in the corpora allata of the last larval instar of Choleva angustata Fab: increase of the glandular volume and development of the smooth reticulum. These findings suggest that fasting leads to a continuous activity of the corpora allata.