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Pedal peptide (PP) and orcokinin (OK) are related neuropeptides that were discovered in protostomian invertebrates (mollusks, arthropods). However, analysis of genome/transcriptome sequence data has revealed that PP/OK-type neuropeptides also occur in a deuterostomian phylum-the echinoderms. Furthermore, a PP/OK-type neuropeptide (starfish myorelaxant peptide, SMP) was recently identified as a muscle relaxant in the starfish Patiria pectinifera. Here mass spectrometry was used to identify five neuropeptides (ArPPLN1a-e) derived from the SMP precursor (PP-like neuropeptide precursor 1; ArPPLNP1) in the starfish Asterias rubens. Analysis of the expression of ArPPLNP1 and neuropeptides derived from this precursor in A. rubens using mRNA in situ hybridization and immunohistochemistry revealed a widespread pattern of expression, with labeled cells and/or processes present in the radial nerve cords, circumoral nerve ring, digestive system (e.g., cardiac stomach) and body wall-associated muscles (e.g., apical muscle) and appendages (e.g., tube feet and papulae). Furthermore, our data provide the first evidence that neuropeptides are present in the lateral motor nerves and in nerve processes innervating interossicular muscles. In vitro pharmacological tests with SMP (ArPPLN1b) revealed that it causes dose-dependent relaxation of apical muscle, tube foot and cardiac stomach preparations from A. rubens. Collectively, these anatomical and pharmacological data indicate that neuropeptides derived from ArPPLNP1 act as inhibitory neuromuscular transmitters in starfish, which contrasts with the myoexcitatory actions of PP/OK-type neuropeptides in protostomian invertebrates. Thus, the divergence of deuterostomes and protostomes may have been accompanied by an inhibitory-excitatory transition in the roles of PP/OK-type neuropeptides as regulators of muscle activity., (© 2017 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc.)

Titania nanoparticles are used in food, cosmetic, medicine, paint and many more domestic items. Its extensive use has raised the threat to the physiological system and thus the functioning of the body. In the current study, the toxicity of TiO 2 is checked by adding it in food and using Drosophila melanogaster as a model organism. Various concentrations of TiO 2 (50, 100, 200, 250mg·L -1 ) toxicity was assessed via oral route exposure. Survivability, life-cycle, mechanosensory behaviour and structure of various mechanosensory organs were monitored as a read out of nanoparticle toxicity. TiO 2 NPs generate reactive oxygen species which can modify multiple signalling pathways and thus can alter the development and behavioural pattern of the fly., (Copyright © 2016 Elsevier Inc. All rights reserved.)

A new method that can improve gas-chromatography-electroantennographic detection (GC-EAD) by orders of magnitude through a technique known as chopper stabilization combined with matched filtering in colored noise is presented. The EAD is a physiological recording from the antenna of an insect which can be used to find compounds in the GC effluent that the antenna is able to detect, having important applications for pest control and understanding of chemical communication in nature. The new method is demonstrated with whole-animal male Helicoverpa zea antennal preparations for detection of major pheromone component (cis-11-hexadecenal) and compared to results obtained using traditional EAD recording techniques. Results indicate that chopper stabilization under these circumstances can increase odorant detection performance by a factor of approximately 10(4) over traditional methods. The time course of the response of the antenna is also better resolved under chopped conditions. Although the degree of improvement is expected to vary with insect species, odor, and insect preparation, under most circumstances a more sensitive and robust GC-EAD instrument will result from the application of this technique., (Copyright © 2011 Elsevier B.V. All rights reserved.)

All scorpions have two mid-ventral organs called pectines. Each pecten has thousands of pore-tipped sensilla sensitive to a variety of volatile organic and water-based stimulants. However, it was previously unknown whether individual sensilla were functionally identical or different. The information enhancement hypothesis predicts that all sensilla have similar chemosensitivities such that each is a unit of a parallel processing system. The information segmentation hypothesis states that sensilla differ in their chemosensitivities, a functional arrangement akin to the glomeruli-specific chemical detection system in the moth or human olfactory sense. In this study, we tested these hypotheses by extracellularly tip-recording sensillar responses to three aqueous tastants: 0.01 M KCl, 0.1 M citric acid, and 40% ethanol by volume. We isolated stimulation to one sensillum at a time and compared the chemoresponses. Sensilla appeared to respond similarly to the same stimulant (i.e., sensillar tip-recordings revealed activity of the same cell types), although sometimes a few sensilla responded with higher spike rates than the others. We conclude that our data primarily support the information enhancement hypothesis but for future tests of sensillar function we suggest a new hybrid model, which proposes that a few specialized sensilla exist among a mostly uniform field of identical sensilla.

Preplacodal ectoderm arises near the end of gastrulation as a narrow band of cells surrounding the anterior neural plate. This domain later resolves into discrete cranial placodes that, together with neural crest, produce paired sensory structures of the head. Unlike the better-characterized neural crest, little is known about early regulation of preplacodal development. Classical models of ectodermal patterning posit that preplacodal identity is specified by readout of a discrete level of Bmp signaling along a DV gradient. More recent studies indicate that Bmp-antagonists are critical for promoting preplacodal development. However, it is unclear whether Bmp-antagonists establish the proper level of Bmp signaling within a morphogen gradient or, alternatively, block Bmp altogether. To begin addressing these issues, we treated zebrafish embryos with a pharmacological inhibitor of Bmp, sometimes combined with heat shock-induction of Chordin and dominant-negative Bmp receptor, to fully block Bmp signaling at various developmental stages. We find that preplacodal development occurs in two phases with opposing Bmp requirements. Initially, Bmp is required before gastrulation to co-induce four transcription factors, Tfap2a, Tfap2c, Foxi1, and Gata3, which establish preplacodal competence throughout the nonneural ectoderm. Subsequently, Bmp must be fully blocked in late gastrulation by dorsally expressed Bmp-antagonists, together with dorsally expressed Fgf and Pdgf, to specify preplacodal identity within competent cells abutting the neural plate. Localized ventral misexpression of Fgf8 and Chordin can activate ectopic preplacodal development anywhere within the zone of competence, whereas dorsal misexpression of one or more competence factors can activate ectopic preplacodal development in the neural plate. Conversely, morpholino-knockdown of competence factors specifically ablates preplacodal development. Our work supports a relatively simple two-step model that traces regulation of preplacodal development to late blastula stage, resolves two distinct phases of Bmp dependence, and identifies the main factors required for preplacodal competence and specification., Competing Interests: The authors have declared that no competing interests exist.

The design principles and specific proteins of the dynein-tubulin motor, which powers the flagella and cilia of eukaryotes, have been conserved throughout the evolution of life from algae to humans. Cilia and flagella can support both motile and sensory functions independently, or sometimes in parallel to each other. In this paper we show that this dual sensory-motile role of eukaryotic cilia is preserved in the most sensitive of all invertebrate hearing organs, the Johnston's organ of the mosquito. The Johnston's organ displays spontaneous oscillations, which have been identified as being a characteristic of amplification in the ears of mosquitoes and Drosophila. In the auditory organs of Drosophila and vertebrates, the molecular basis of amplification has been attributed to the gating and adaptation of the mechanoelectrical transducer channels themselves. On the basis of their temperature-dependence and sensitivity to colchicine, we attribute the molecular basis of spontaneous oscillations by the Johnston's organ of the mosquito Culex quinquefasciatus, to the dynein-tubulin motor of the ciliated sensillae. If, as has been claimed for insect and vertebrate hearing organs, spontaneous oscillations epitomize amplification, then in the mosquito ear, this process is independent of mechanotransduction.

Olfactory perception of myristic, palmitic, stearic and oleic acids and their corresponding methyl esters by Asian corn borer moths, Ostrinia furnacalis (Guenée) (Lepidoptera: Crambidae) was investigated. It was found that mated females with both antennae amputated, in contrast to intact females and females with one antenna removed, could not discriminate between simultaneously provided control filter papers and filters treated with a blend of oviposition-deterring fatty acids. Oviposition by mated females exhibited a very marked periodicity, with all egg masses deposited during the scotophase and most egg masses laid before midnight. According to the peak and trough period of oviposition, electroantennogram (EAG) responses from both mated females and males to the four fatty acids and four methyl esters were tested within two two-hour periods from 3 to 5 hours after the start of darkness and from 1 to 3 hours after light onset, respectively. Significant EAG responses above solvent and background were elicited by all test chemicals from females, and by most of the test compounds from males. EAG values of all test chemicals from mated females were not statistically different between the two test periods except for methyl myristate. Conversely, EAG responses from mated males to myristic acid, stearic acid and their methyl esters significantly differed between the two test periods.

An electrogenic K(+) transport in the tormogen cell of insect chemosensilla is involved in the generation and maintenance of the transepithelial potential (TEP). To gain more information about the K(+) transport system underlying the TEP generation and the location of its components in the plasma membrane of the tormogen cell, we studied the effects of inhibitors of K(+)/H(+) P-ATPase (bafilomycin A1, omeprazole and Na-orthovanadate), of K(+)/Cl(-) co-transport (bumetanide), of Cl(-) channels (NPPB) and of a K(+) channel blocker (BaCl(2)). The relationship between TEP amplitude and spike firing activity was also studied. Experiments were performed on the labellar chemosensilla of the blowfly Protophormia terraenovae using a modified tip-recording technique. Results show that: (a) K(+)/H(+) P-ATPase inhibitors significantly decrease the TEP, when properly applied to the labellum for 20 min, so as to reach the basolateral side of the plasma membrane, while no effect was detected when applied to the apical side, (b) bumetanide, NPPB and BaCl(2) decrease the TEP value only when administered to the apical side, (c) spike activity is positively correlated with the TEP. A model is proposed of the active and passive K(+) transports sustaining the TEP associated with the blowfly chemosensilla.

The electrophysiological response of labellar and tarsal chemosensilla in the blowfly Phormia regina was studied in response to a complex stimulus naturally encountered by flies such as sheep faeces, and to beef liver, a proteinaceous feeding source. Responses were investigated both before or after injection of clonidine, an octopamine agonist previously shown to enhance sucrose ingestion, while decreasing that of proteins. As assessed by single sensillum recordings, the four different chemosensory - "salt", "sugar", "deterrent" and "water" - cells were all activated by both stimuli, regardless of sex and sensillum type, the "sugar" one being in all cases the most sensitive to beef liver before clonidine injection. Clonidine treatment affected neither labellar nor tarsal sensitivity to sucrose. Conversely, clonidine-injected flies showed a significant increase in the activity of the "deterrent" cell to beef liver, thus accounting for a decrease in protein ingestion. This study for the first time provides evidence of a key role of a clonidine-sensitive peripheral taste sensitivity in down-regulation of protein ingestion in blowflies. Correlation between peripheral sensitivity and behavioural output is discussed.

The nervous system copes with variability in the external and internal environment by using neuromodulators to adjust the efficacy of neural circuits. The role of serotonin (5HT) as a neuromodulator of olfactory information processing in the antennal lobe (AL) of Manduca sexta was examined using multichannel extracellular electrodes to record the responses of ensembles of AL neurons to olfactory stimuli. In one experiment, the effects of 5HT on the concentration-response functions for two essential plant oils across a range of stimulus intensities were examined. In a second experiment, the effect of 5HT on the ability of ensembles to discriminate odorants from different chemical classes was examined. Bath application of 5HT enhanced AL unit responses by increasing response duration and firing rate, which in turn increased the amount of spike time cross-correlation and -covariance between pairs of units. 5HT had the greatest effect on overall ensemble activation at higher odorant concentrations, resulting in an increase in the gain of the dose-response function of individual units. Additionally, response thresholds shifted to lower odorant concentrations for some units, suggesting that 5HT increased their sensitivity. Serotonin enhanced ensemble discrimination of different concentrations of individual odorants as well as discrimination of structurally dissimilar odors at the same concentration. Given the known circadian fluctuations of 5HT in the AL of this species, these findings support the hypothesis that 5HT periodically enhances sensitivity and responsiveness in the AL of Manduca to maximize efficiency when the requirement for olfactory acuity is the greatest.

It is known that human individuals show different levels of attractiveness to mosquitoes. In this study, we investigated the chemical basis for low attractiveness. We recorded behaviors of Aedes aegypti toward the hands of human volunteers and toward the volatile chemicals produced by their bodies. Some individuals, and their corresponding volatiles, elicited low upwind flight, relative attraction, and probing activity. Analyzing the components by gas chromatography coupled to electrophysiological recordings from the antennae of Aedes aegypti, enabled the location of 33 physiologically relevant compounds. The results indicated that higher levels of specific compounds may be responsible for decreased "attractiveness." In behavioral experiments, five of the compounds caused a significant reduction in upwind flight of Aedes aegypti to attractive human hands. Thus, unattractiveness of individuals may result from a repellent, or attractant "masking," mechanism.

Pheromones are known to be important to the innate behavior of marine animals. Attraction in Aplysia involves the long-distance water-borne protein pheromones attractin, enticin, temptin and seductin, which are released from the albumen gland during egg laying. Other pheromones are predicted to act in concert with these pheromones, but their identities are unknown. To identify additional pheromone candidates, we employed differential library screening of an albumen gland cDNA library, RT-PCR, recombinant protein expression, rhinophore contraction bioassays and immunocytochemistry. Alb-1 is expressed in the Aplysia californica albumen gland and encodes a novel protein that does not share significant sequence identity with any proteins in the database. RT-PCR analysis detected Alb-1 transcripts in the albumen gland, exocrine atrial gland and ovotestis. The Alb-1 precursor has a signal peptide sequence followed by a predicted 101-residue protein sequence containing eight cysteine residues. Recombinant protein expression, RP-HPLC, microsequence analysis and MALDI mass spectrometry analyses demonstrated that mature recombinant Alb-1 was processed at a paired basic residue site to generate an N-terminal and C-terminal protein fragment; this was consistent with immunoblot observations on purified albumen gland extracts. In rhinophore contraction (twitch) bioassays, the recombinant N-terminal protein induced rhinophore contractions whereas the C-terminal protein did not. An antibody generated to the N-terminal protein was used for immunocytochemical and immunoblot analyses and demonstrated that this protein is present in albumen gland secretory cells, egg cordons and egg eluates. Overall, the data suggest that Alb-1 may be processed in the albumen gland and that the Alb-1 1-56 protein released during egg laying may serve a pheromonal function in concert with attractin, enticin, temptin and seductin.

In the caddisfly Molanna angustata, females produce a sex pheromone in glands with openings on the fifth sternite. Gas chromatographic analyses of pheromone gland extracts with electroantennographic detection revealed four major compounds that stimulated male antennae. These compounds were identified by means of gas chromatography-mass spectrometry and enantioselective gas chromatography as heptan-2-one, (S)-heptan-2-ol, nonan-2-one, and (S)-nonan-2-ol in the approximate ratio of 1:1:4:10, respectively. Field tests showed that the mixture of the two alcohols was attractive to males whereas addition of the corresponding ketones reduced trap catches. The sex pheromone of M. angustata, a species in the family Molannidae within the suborder Integripalpia, is similar to the pheromones or pheromone-like compounds previously reported from six other trichopteran families, including members of the basal suborder Annulipalpia. This suggests that minimal evolutionary change of the pheromone chemistry has taken place within the leptoceroid branch of integripalpian Trichoptera compared to the ancestral character state.

Inflorescence scent samples from nine populations of dioecious Silene otites, a plant pollinated by moths and mosquitoes, were collected by dynamic headspace extraction. Sixty-three scent samples were analyzed by gas chromatography-mass spectrometry. Out of 38 found, 35 compounds were identified, most of which were monoterpenoids, fatty acid derivatives, and benzenoids. Phenyl acetaldehyde was the most dominant compound in the majority of samples. The variability in scent composition was high, and population and sex differences were found. Nevertheless, wind tunnel experiments proved similar attraction of Culex pipiens pipiens biotype molestus mosquitoes to the inflorescence odor of S. otites of different populations, indicating that different blends are similarly attractive to mosquitoes. The electrophysiological responses of mosquitoes to the 12 most common and abundant odor compounds of S. otites differed. Linalool oxide (furanoid) and linalool evoked the strongest responses in male and female mosquitoes, and (Z)-3-hexenyl acetate was strongly active in females. Medium responses were evoked in males by (Z)-3-hexenyl acetate, in females by benzaldehyde and methyl salicylate, and in both sexes by lilac aldehyde, lilac alcohol, and linalool oxide (pyranoid).

While testing traps baited with a blend of geranyl octanoate and geranyl butanoate (pheromone components previously identified for Agriotes lineatus, Coleoptera, Elateridae) in Portugal and Bulgaria, large numbers of the closely related Agriotes proximus were captured. In the literature, two different compounds, (E,E)-farnesyl acetate and neryl isovalerate had previously been identified as pheromone components of A. proximus. Subsequent field tests, conducted in several European countries, revealed that A. proximus was weakly attracted to geranyl butanoate on its own, while A. lineatus was weakly attracted to geranyl octanoate on its own. However, the largest catches for both species were observed with a blend of both compounds. No A. proximus was caught in traps baited with the blend of (E,E)-farnesyl acetate and neryl isovalerate at any of the test sites. In electroantennographic studies, antennae of male A. proximus and A. lineatus both gave greater responses to geranyl butanoate than to geranyl octanoate, suggesting that the perception of these two compounds was similar for both species. A 1:1 blend of geranyl octanoate and geranyl butanoate can be used as a bait in traps for the detection and monitoring of both A. lineatus and A. proximus in many European countries.

To identify the sensory organs that are sensitive to water stimuli in the cricket Gryllus bimaculatus, cuticular structures on the legs and the number of sensory neurons innervating them were studied. Some small hair sensilla on the legs were innervated by 2-5 sensory neurons. All such sensilla had a tiny pore at the tip of their hairs. The diameter of the pore was approximately 0.2 mum. These findings suggest that these are chemosensitive hairs (LCS: leg chemosensillum). Of the three pairs of legs, the anterior legs (forelegs) possessed the largest number of LCSs. Of the five leg segments (i.e., coxa, trochanter, femur, tibia and tarsus), the tarsus possessed the largest number of LCSs on each leg. Electrophysiological investigation by tip recording revealed that some of the LCSs contained water-receptor cells. Because the basitarsus possessed a larger number of LCSs than the other tarsomeres, the distribution of water-receptor-containing LCSs in the basitarsus of a foreleg was investigated morphologically and electrophysiologically. LCSs that contained water-receptor cells were mainly distributed on the ventral surface of the basitarsus. There were two types of water receptor that showed different response patterns to a stimulus, that is, phasic- and tonic-type water receptors. From the distribution of LCSs on the legs, the roles of these different types of water receptors in behavioral selection, that is, the initiation of swimming and the inhibition of flying, will be discussed.

Taste is an essential sense for detection of nutrient-rich food and avoidance of toxic substances. The Drosophila melanogaster gustatory system provides an excellent model to study taste perception and taste-elicited behaviors. "The fly" is unique in the animal kingdom with regard to available experimental tools, which include a wide repertoire of molecular-genetic analyses (i.e., efficient production of transgenics and gene knockouts), elegant behavioral assays, and the possibility to conduct electrophysiological investigations. In addition, fruit flies, like humans, recognize sugars as a food source, but avoid bitter tasting substances that are often toxic to insects and mammals alike. This paper will present recent research progress in the field of taste and contact pheromone perception in the fruit fly. First, we shall describe the anatomical properties of the Drosophila gustatory system and survey the family of taste receptors to provide an appropriate background. We shall then review taste and pheromone perception mainly from a molecular genetic perspective that includes behavioral, electrophysiological and imaging analyses of wild type flies and flies with genetically manipulated taste cells. Finally, we shall provide an outlook of taste research in this elegant model system for the next few years.

Background: Insects detect a multitude of odors using a broad array of phenotypically distinct olfactory organs referred to as olfactory sensilla. Each sensillum contains one to several sensory neurons and at least three support cells; these cells arise from mitotic activities from one or a small group of defined precursor cells. Sensilla phenotypes are defined by distinct morphologies, and specificities to specific odors; these are the consequence of developmental programs expressed by associated neurons and support cells, and by selection and expression of subpopulations of olfactory genes encoding such proteins as odor receptors, odorant binding proteins, and odor degrading enzymes., Methodology/principal Findings: We are investigating development of the olfactory epithelium of adult M. sexta, identifying events which might establish sensilla phenotypes. In the present study, antennal tissue was examined during the first three days of an 18 day development, a period when sensory mitotic activity was previously reported to occur. Each antenna develops as a cylinder with an outward facing sensory epithelium divided into approximately 80 repeat units or annuli. Mitotic proliferation of sensory cells initiated about 20-24 hrs after pupation (a.p.), in pre-existing zones of high density cells lining the proximal and distal borders of each annulus. These high density zones were observed as early as two hr. a.p., and expanded with mitotic activity to fill the mid-annular regions by about 72 hrs a.p. Mitotic activity initiated at a low rate, increasing dramatically after 40-48 hrs a.p.; this activity was enhanced by ecdysteroids, but did not occur in animals entering pupal diapause (which is also ecdysteroid sensitive)., Conclusions/significance: Sensory proliferation initiates in narrow zones along the proximal and distal borders of each annulus; these zones rapidly expand to fill the mid-annular regions. These zones exist prior to any mitotic activity as regions of high density cells which form either at or prior to pupation. Mitotic sensitivity to ecdysteroids may be a regulatory mechanism coordinating olfactory development with the developmental choice of diapause entry.

The response threshold hypothesis of division of labour in honey bees assumes that individuals differ in their responsiveness to different stimulus modalities. However, previous experiments have shown that responsiveness to gustatory stimuli correlates with responsiveness to odours, pollen and tactile stimuli. Evaluation of these stimuli involves sensory receptors on the antenna. We tested whether responsiveness to gustatory stimuli correlates with responsiveness to visual stimuli in a phototaxis experiment, which is independent of antennal input. Gustatory responsiveness was measured using the proboscis extension response to antennal stimulation with water and different sucrose concentrations. Phototaxis was quantified by measuring the walking times a bee needed to reach light sources of different intensities. Walking behaviour in the darkness was measured to test for differences in locomotor behaviour. The walking time towards a light stimulus, the path length, and the walking speed depended on the intensity of the light stimulus. Responsiveness to visual stimuli correlated significantly with gustatory responsiveness. Bees displaying a high gustatory responsiveness were also very sensitive to light. Locomotor activity did not correlate with gustatory responsiveness. This shows that gustatory responsiveness is a good indicator of sensitivity for visual stimuli, which are not perceived by the antenna.

Two sibling species, Helicoverpa assulta and Helicoverpa armigera both use (Z)-9-hexadecenal and (Z)-11-hexadecenal as their sex pheromone components but in almost reversed ratios, 93:7 and 3:97, respectively. H. assulta and H. armigera males performed upwind flight in response to the H. assulta sex pheromone blend (93:7). H. armigera responded strongly to the H. armigera blend (3:97), whereas H. assulta males remained inactive upon exposure to this blend. Both species gave clear dose-dependent electrophysiological responses to (Z)-11-hexadecenal. However, (Z)-9-hexadecenal evoked strong dose-dependent electrophysiological responses in H. assulta males but not in H. armigera. The two male F1 hybrids exhibited similar behavioral responses to two sex pheromone blends and electrophysiological responses to two pheromone components as H. armigera males. This indicated that H. armigera genes appear dominant in determining the behavioral response and electrophysiological responses. Behavioral and electrophysiological responses of backcrosses of male F1 hybrids (H. armigera female x H. assulta male) with female H. assulta and H. armigera were close to that of H. assulta and H. armigera, respectively. However, backcrosses of female F1 hybrids (H. assulta female x H. armigera male) with male H. assulta and H. armigera showed reduced behavioral responses but normal electrophysiological responses compared to males of the respective parental line.

The primordial functional role of honey bee males (drones) is to mate with virgin queens, a behaviour relying heavily on the olfactory detection of queen pheromone. In the present work I studied olfactory processing in the drone antennal lobe (AL), the primary olfactory centre of the insect brain. In drones, the AL consists of about 103 ordinary glomeruli and four enlarged glomeruli, the macroglomeruli (MG). Two macroglomeruli (MG1 and MG2) and approximately 20 ordinary glomeruli occupy the anterior surface of the antennal lobe and are thus accessible to optical recordings. Calcium imaging was used to measure odour-evoked responses to queen pheromonal components and plant odours. MG2 responded specifically to the main component of the queen mandibular pheromone, 9-ODA. The secondary components HOB and HVA each triggered activity in one, but not the same, ordinary glomerulus. MG1 did not respond to any of the tested stimuli. Plant odours induced signals only in ordinary glomeruli in a combinatorial manner, as in workers. This study thus shows that the major queen pheromonal component is processed in the most voluminous macroglomerulus of the drone antennal lobe, and that plant odours, as well as some queen pheromonal components, are processed in ordinary glomeruli.

The aim of this study was to detect the role of some mosquito organs in their sensation of repellent materials. A total of 250 females (15 days old) of the target species Aedes aegypti and Anopheles stephensi were prepared and divided into five groups: group 1, without antenna; group 2, without maxillary bulbs; group 3, without proboscis; group 4, without frontal tarsus; and group 5, normal females as control. A mixture of five oils containing Litsea cubeba 1%, Melaleuca leucadendron 1%, Melaleuca quinquenervia 1%, Viola odorata 1%, and Nepeta cataria 1% was included in a complex solvent containing 20% genapol, 10% polyethylene glycol, 20% ethanol, and 50% water. Furthermore, Bayrepel was used in this experiment at a 20% concentration in the same solvent. Pure water was used as control in this study. The test was carried out by spreading 100 microl of the repellent material or water on a 30-cm2 exposure area of a human volunteer's arm. In A. aegypti, the biting and landing percentages increased significantly in those mosquito groups that lacked some organs (especially maxillary bulbs), while in A. stephensi, it became not clear which organ is responsible for perception of repellents.

The effects of anesthesia with etomidate on the cellular mechanisms of sensory processing and sensorimotor coordination have been studied in the active electric sense of the mormyrid fish Gnathonemus petersii. Like many anesthetics, etomidate is known to potentiate GABA(A) receptors, but little is known about the effects on sensory processing at the systems level. A better understanding is necessary for experimental studies of sensory processing, in particular regarding possible effects on the dynamic structure of excitatory and inhibitory receptive fields and to improve the knowledge of the mechanisms of anesthesia in general. Etomidate slowed the electromotor discharge rhythm, probably because of feedback inhibition at the premotor level, but did not alter the structure of the electromotor command. Sensory translation through primary afferents projecting to the cerebellum-like electrosensory lobe (ELL) was not changed. However, central interneurons and projection neurons were hyperpolarized under etomidate, and their spiking activity was reduced. Although the spatial extent and the center/surround organization of sensory receptive fields were not changed, initial excitatory responses were followed by prolonged inhibition. Corollary discharge input to ELL was maintained, and the temporal sequence of excitatory and inhibitory components of this descending signal remained intact. Later inhibitory corollary discharge responses were prolonged by several hundred milliseconds. The result was that excitatory reafferent sensory input was conserved with enhanced precision of timing, whereas background activity was greatly reduced. Anti-Hebbian synaptic plasticity evoked by association of sensory and corollary discharge input was still present under anesthesia, and differences compared with the nonanesthetized condition are discussed.

Analyses of sediment and water indicate the presence of heavy metal pollutants like lead, zinc, copper, mercury and cadmium of the river Damodar of India. These metals are responsible for causing morphological deformities of antennae and other parts of chironomid larvae. Percentage of deformity correlated positively with the concentrations of Pb in water and sediment (r > 0.6) at the confluence point. A new severity index, SISS((antenna)) has been proposed here to assess deformity at the family or subfamily level.

The sex pheromone of the German cockroach, Blattella germanica, has been characterized as gentisyl quinone isovalerate. This cockroach is a major cause of allergic disease and serves as a mechanical vector of pathogens, making it one of the most important residential and food-associated pests worldwide. The sex pheromone-producing gland in adult females was identified in 1993, but thermal instability of the pheromone made characterization difficult. Now, using a new preparative gas chromatography approach coupled with electroantennographic detection, we have isolated and characterized the pheromone, which we term blattellaquinone, and confirmed the identification by chemical synthesis. The synthetic pheromone was active in behavioral assays and highly effective in field trapping tests, which suggest that it may provide a new tool in cockroach population detection, monitoring, and control.

Larvae of the cabbage white Pieris rapae are specialists on plants belonging to the family Brassicaceae (Cruciferae). Adult females have been shown to use the glucosinolate gluconasturtiin (phenylethylglucosinolate) as a recognition cue for cruciferous plants, so they can identify an appropriate host for oviposition (Huang and Renwick in J Chem Ecol 20:1025-1037, 1994). Here, we report our results from a study of the role of this glucosinolate in feeding preferences of P. rapae larvae. The larvae were allowed to choose between leaf disks from the non-host cowpea Vigna sinensis (Fabaceae) that were treated with pure gluconasturtiin in solvent, or solvent alone. Our results showed that gluconasturtiin is a feeding stimulant for P. rapae larvae. A series of chemosensory ablations revealed that this response is mediated by one set of taste sensilla, the sensilla styloconica. Electrophysiological tip recordings revealed two neurons in the lateral sensillum styloconicum that are sensitive to gluconasturtiin. These neurons show significantly higher firing frequencies with 4 mM gluconasturtiin added to the recording pipette than for recording solution alone. We propose that the sensitivity to gluconasturtiin shown by these two taste neurons is an important contributor to the animals' behavioral preference for this compound.

The first recorded electroantennographic preliminary studies on stingless bees have been performed using two species of Frieseomelitta from Brazil. Experiments with F. silvestrii and F. varia showed that antennae respond to hexane extracts of heads and abdomens of both species and posterior tibia of F. silvestrii (which carry plant resin), as well as to the pure compounds 2-heptanol and 2-nonanol, which occur in the mandibular glands of both species, and to the terpenes alpha-cubebene, humulene, and beta-caryophyllene found on their tibia and in the cerumen of their nests.

The results of experiments on regeneration of the Haller's sensory organ in the metastriate ixodid tick Haemaphysalis longicornis Neumann in the presence of the juvenoid methoprene and retinoic acid and of studies of its structural changes by SEM confirmed the similarity of prostriate and metastriate ixodids as concerns the juvenalizing effect of the above morphogens on regenerative processes during nymphal-imaginal metamorphosis. However, the metastriate ticks (Haemaphysalis and earlier studied Hyalomma) are well behind the prostriate ticks (Ixodes) as concerns the extent of changes induced by juvenoids and retinoids in the sensillar sets of Haller's organ regenerates.

The primary olfactory centre, the antennal lobe of Heliothis virescens moths, contains 62 glomeruli which process plant odour information and four male-specific glomeruli which form the macroglomerular complex, involved in processing information about pheromone and interspecific signals. Using calcium imaging, we recorded the spatio-temporal activity pattern of the glomeruli in the anterior antennal lobe during stimulation with odorants produced by plants or insects. Each odorant elicited specific excitatory responses in one or a few glomeruli: the major pheromone component did so exclusively in the large glomerulus of the macroglomerular complex and the plant odours exclusively in the ordinary glomeruli. Eight glomeruli, with corresponding plant odour responses and positions, were identified within each sex. Glomeruli responded specifically to linalool, beta-ocimene/beta-myrcene or germacrene D/alpha-farnesene. Responses to two essential plant oils covered the response areas of their major constituents, as well as activating additional glomeruli. Stronger activation in the AL due to increased odour concentration was expressed as increased response strength within the odorant-specific glomeruli as well as recruitment of less sensitive glomeruli.

In the attempt to gain more information on the mechanisms underlying bitter and/or sweet taste reception, we have investigated the responses of labellar chemosensilla in the blowfly Protophormia terraenovae to Na-saccharin, as compared to sweet stimuli (sucrose or fructose) and bitter stimuli (denatonium benzoate or amiloride). Electrophysiological and behavioral results indicate that the sweetener Na-saccharin inhibits the "sugar" cell in the labellar taste sensilla of the blowfly P. terraenovae. In multichoice preference tests, flies ingested more of the solutions containing sugar to those with sugar+Na-saccharin. This finding is in good agreement with the spike frequency reduction observed for the "sugar" cell activity. Analysis of the spike discharges also shows a positive dose-response for the "deterrent" cell following stimulation with Na-saccharin and denatonium benzoate. Flies drank any of the Na-saccharin solutions, regardless of their concentration, less than water, thus indicating a weak deterring effect on water drinking. The prevailing activation of the "deterrent" cell by stimulation with Na-saccharin is not directly coupled with a coherent behavioral output. Cross adaptation was found to occur between responses to Na-saccharin and denatonium benzoate or amiloride regardless of the order of adapting stimuli. In the case of sweet stimuli, cross adaptation occurred when the adapting stimulus was Na-saccharin, but it did not when the adapting stimuli were sucrose or fructose. Addition of Na-saccharin to both sugars significantly depressed the spike firing frequency, while an increase was observed with denatonium benzoate or amiloride.

We investigated strategies involved in odour intensity coding by the primary olfactory centre of insects, the antennal lobe (AL), the structural and functional analogue of the olfactory bulb. Using calcium imaging in the honeybee, we simultaneously measured the projection neuron output responses and a compound signal dominated by receptor neuron input in identified olfactory glomeruli to odours spanning seven log units of concentration. A comparison of the two processing levels indicates that the intercellular computation within the AL modulates and contrast-enhances the primary olfactory signals. As a result the AL network optimizes the olfactory code: odour representation is improved at lower concentrations, the relative activity of olfactory glomeruli allows encoding odour quality over up to four log-unit concentrations, and odour-intensity is reliably represented in the overall excitation across AL.

Specificity of olfactory receptor neurones plays an important role in food and host preferences of a species, and may have become conserved or changed in the evolution of polyphagy and oligophagy. We have identified a major type of plant odour receptor neurones responding to the sesquiterpene germacrene D in three species of heliothine moths, the polyphagous Heliothis virescens and Helicoverpa armigera and the oligophagous Helicoverpa assulta. The neurones respond with high sensitivity and selectivity to (-)-germacrene D, as demonstrated by screening via gas chromatography with numerous mixtures of plant volatiles. Germacrene D was present in both host and non-host plants, but only in half of the tested species. The specificity of the neurones was similar in the three species, as shown by the "secondary" responses to a few other sesquiterpenes. The effect of (-)-germacrene D was about ten times stronger than that of the (+)-enantiomer, which again was about ten times stronger than that of (-)-alpha-ylangene. Weaker effects were obtained for (+)-beta-ylangene, (+)-alpha-copaene, beta-copaene and two unidentified sesquiterpenes. The structure-activity relationship shows that the important properties of (-)-germacrene D in activating the neurones are the ten-membered ring system and the three double bonds acting as electron-rich centres, in addition to the direction of the isopropyl-group responsible for the different effects of the germacrene D enantiomers.

Like many other complex biological phenomena, sensory processing is starting to be studied at the level of the gene. Whereas the mouse is the standard organism for molecular neurobiology, somatosensation and pain are mainly investigated in the rat. The in-vitro electrophysiological technique is well established in mice. The effect of manipulations of the mouse genome on neuronal systems and networks has to be investigated under in-vivo conditions. Thus, the step from the rat to the smaller mouse seems to be important in terms of progress in molecular neurobiology. So far, there is no published study of sensory processing by electrophysiological means in mice under controlled general anesthesia. In this paper we describe a technique for electrophysiological recordings of sensory and sensorimotor processing in anesthetized mice. The mouse is anesthetized by continuous administration of the short-acting general anesthetic methohexital sodium salt via the jugular vein and artificially ventilated via a tracheotomy. The electrocardiogram and the carotid artery blood pressure are monitored. The sensorimotor jaw-opening reflex and the hindpaw withdrawal reflex are elicited by electric stimulation of the tongue and the sole of the hindpaw, respectively. The jaw-opening reflex is inhibited by deep brain stimulation of the rostral ventromedial medulla. On- and off-cells of the rostral ventromedial medulla and sensory neurons of the spinal trigeminal nucleus are extracellularly recorded by tungsten electrodes. General anesthesia can be reliably induced, maintained and sufficiently controlled in mice. Under this condition, studies can be performed on sensory processing in mice.

Odours are represented as unique combinations of activated glomeruli in the antennal lobes of insects. Receptor neurons arborizing in the glomeruli are not only qualitatively selective, but in addition respond to variations in stimulus concentration. As each glomerulus likely represents a single receptor neuron type, optical recordings of calcium changes in insect antennal lobes show how concentration variations affect a large population of afferents. We measured the glomerular responses in the moth Spodoptera littoralis to different concentrations of plant-related odorants. Localized calcium responses were shown to correspond to individual glomeruli. We found that the dynamic range of glomerular responses spanned 3-4 log units of concentration and the most strongly responding glomeruli often reached a plateau at high stimulus doses. Further, we showed that the single most active glomerulus was often not the same across concentrations. However, if the principal glomerulus moved, it was generally to an adjacent or proximal glomerulus. As concentration increased, a higher number of glomeruli became activated. Correlations of glomerular representations of the same compound at different doses decreased as the difference in concentration increased. Moreover, representations evoked by different odorants were more correlated at high than at low doses, which means that the uniqueness of activity patterns decreased with increasing concentration. Thus, if odours are coded as spatial patterns of glomerular activity, as has been suggested, these olfactory codes are not persistent across concentrations.

Serotonin (5-HT) is a neurotransmitter which is supposed to play a key role during development. In the last few years 5-HT receptors have been cloned in many animal species, and there is evidence that different 5-HT receptors are also present in ascidians. Ascidians and vertebrates are both members of the phylum Chordata and both have a dorsal tubular central nervous system. Embryos of the ascidian Phallusia mammillata have been treated with WAY-100635, a potent and selective 5-HT(1A) receptor antagonist. The larvae developed from treated embryos showed a dramatic reduction of their anterior sensory vesicles and the pigment of two sensory organs, the ocellus and the otolith. Immunofluorescence experiments with an anti beta-tubulin monoclonal antibody specific for the neural system showed that the anterior neural system of treated animals was radically altered by the action of the drug in a dose-dependent way. These results suggest that 5-HT plays a role in the development of the neural system in ascidians and its action is mediated by receptors similar to the members of the 5-HT(1A) receptor subtype of mammals.

Background: Hemorrhagic shock produces an immediate activation of the autonomic nervous system and endogenous opioid pathways. Our studies have demonstrated that endogenous opioid activation aggravates the hemodynamic and inflammatory responses to shock. However, it is unclear whether endogenous opioid activation is triggered by noxious stimuli and furthermore whether it produces analgesia., Methods: Experiments were conducted in chronically catheterized, conscious, unrestrained, nonheparinized, male, Sprague-Dawley rats subjected to fixed pressure hemorrhage. Blood samples were obtained for determinations of circulating beta-endorphin and substance P. Analgesia was measured using the tail-flick response to a noxious stimulus before and during hemorrhage. The contribution of sensory neurons to eliciting the neuroendocrine, opioid, and inflammatory responses to hemorrhage was investigated in capsaicin-treated animals., Results: Hemorrhagic shock produced marked naltrexone-sensitive analgesia without significant modulation of substance P. Peripheral sensory denervation did not alter the hemodynamic, neuroendocrine, or inflammatory responses to shock., Conclusion: Endogenous opioid activation during shock produces analgesia. Sensory neuron activation appears to have limited effect on shock-induced hemodynamic and proinflammatory responses. Furthermore, these results suggest that the activation of neuroendocrine and opioid pathways during shock is not likely to be a response to noxious stimuli.

The sphinx moth Manduca sexta is a well-studied insect with regard to central olfactory functions. Until now, the innervation patterns of olfactory receptor neurons into the array of olfactory glomeruli in the antennal lobe have, however, been unclear. Using optical imaging to visualize calcium dynamics within the antennal lobe we demonstrate specific patterns elicited by sex pheromone components and plant-derived odours. These patterns mainly reflect receptor neuron activity. Within the male-specific macroglomerular complex the two major pheromone components evoke stereotyped activity in either of two macroglomerular complex glomeruli. Based on previous knowledge of output neuron specificity, our results suggest a matching of information between input and output in the macroglomerular complex. Plant odours evoked activity in the sexually isomorphic glomeruli. Two major results were obtained: (1). terpenes and aromatic compounds activate different clusters of glomeruli with only minor overlapping, and (2). the position of certain key glomeruli is fixed in both males and females, which suggests that host-plant related odorants are processed in a similar way in both sexes.

Effects of 20-hydroxyecdysone and serotonin on the morphological development and the survival of antennal lobe neurons from day-2 pupal brains of the silk moth Bombyx mori were investigated in vitro. Four morphologically distinct neuronal types could be identified in the cultured antennal lobe neurons: unipolar, bipolar, multi-polar and projection neurons. Antennal lobe neurons in culture with 20-hydroxyecdysone and serotonin showed different patterns of the morphological development from those described in Manduca sexta. Projection neurons extend their neurites remarkably by 20-hydroxyecdysone in B. mori, but there is no extension from antennal lobe neurons in M. sexta. Multi-polar neurons conspicuously increase only formation of new branches from their primary neurites by serotonin in B. mori, but there are both extension and branching of the neurites in M. sexta. On day-5, antennal lobe neurons in lower titers of 20-hydroxyecdysone had significantly higher survival rates than those in higher titers. Neurons cultured for 7 days at different levels of 20-hydroxyecdysone generally showed significantly lower survival rates than neurons cultured for 5 days under the same conditions.

Proteins isolated from the host cocoon of Acrolepiopsis assectella (Lepidoptera: Yponomeutoidea) act as kairomones for host acceptance by the endoparasitoid wasp Diadromus pulchellus Wesmael (Hymenoptera: Ichneumonidae). In this study, morphological, ultrastructural and electrophysiological studies were carried out in order to identify the contact chemoreceptive sensilla on the parasitoid antennae that perceive the protein kairomones. Three types of sensillum on the antennae of the females were found to have a chemosensory function. The receptor cell(s) of one sensillar type were shown to give a positive electrophysiological response to protein kairomones. This sensillar type is apically multiporous and female specific. Consequently, this sensillum could be the one implicated in the perception of the protein kairomone that triggers the host-acceptance behaviour of D. pulchellus females.

Many smokers choose menthol-flavored cigarettes, however, the influence of menthol on the effects of smoke-delivered nicotine is unknown. Research and commercial cigarettes, menthol and non-menthol, that delivered a wide range of nicotine were evaluated. Menthol (n=18) and non-menthol (n=18) cigarette smokers participated in a single session during which three cigarettes were smoked 45 min apart, in random order. Federal Trade Commission (FTC) nicotine yields of the three cigarettes were: research, low yield, 0.2 mg, commercial cigarettes (average), 1.2 mg; research, high yield, 2.5 mg. Commercial and high-yield cigarettes increased heart rate (HR) and blood pressure more than low-yield cigarettes; although, no differences in exhaled carbon monoxide (CO) occurred. Participants smoked commercial cigarettes faster and with fewer puffs than either of the research cigarette indicating production differences can affect topography. There was a significant group by cigarette interaction on satisfaction, and relief from cigarette craving. High-yield non-menthol cigarettes reduced craving and were rated as more satisfying than high-yield menthol cigarettes. No differences between menthol and non-menthol cigarettes on other subjective measures (strength, psychological reward, negative effects) were observed. Our findings indicate that nicotine delivery, but not mentholation, influences cardiovascular and most subjective measures. These results illustrate the importance of threshold levels of nicotine on subjective responses to cigarette smoking.

csp, a gene encoding a protein with high sequence identity to trypsinlike serine protease and CUB domains, was identified from a cDNA library from the olfactory organ (antennular lateral flagellum) of the spiny lobster Panulirus argus. The full-length cDNA sequence of csp is 1801 bp, encoding a protein of 50.25 kD, with three domains: signal peptide, trypsinlike serine protease, and CUB (named for a class of compounds including Complement subcomponents Clr/Cls, Uegf, and Bone morphogenic protein-1). RT-PCR, Northern blots, and immunoblots showed that csp is predominantly expressed in the lateral flagellum and eyestalk. Immunocytochemistry showed that Csp is present in olfactory (aesthetasc) sensilla around auxiliary cells (glia that surround the inner dendrites of olfactory receptor neurons, ORNs) and ORN outer dendrites. We propose that Csp is expressed and secreted by auxiliary cells, associates with ORN cell membranes or extracellular matrix via the CUB domain, and has trypsinlike activity. In the eyestalk, Csp is associated with cells surrounding axons between neuropils of the eyestalk ganglia. Possible functions in the olfactory organ and eyestalk are discussed. To our knowledge, this is the first report from any olfactory system of a gene encoding a protein with serine protease and CUB domains., (Copyright 2001 John Wiley & Sons, Inc.)

Reliable retention of olfactory learning following a 1-trial classical conditioning of the proboscis extension reflex (PER) is not achieved in honeybees until they are 6-7 days old. Here we show that treatment of newly emerged honeybees with juvenile hormone (JH) has a profound effect on the maturation of short-term olfactory memory. JH-treated individuals display excellent short-term (1 h) memory of associative learning at times as early as 3 days of age and perform consistently better than untreated bees for at least the first week of their lives. By contrast, the retention of long-term (24 h) memory following a 3-trial conditioning of the PER is not significantly improved in JH-treated bees. Our study also shows that experience and (or) chemosensory activation are not essential to improve learning performance in olfactory tasks. The lack of accelerated development of long-term retention of olfactory memories in JH-treated honeybees is discussed in the context of neural circuits suspected to mediate memory formation and retrieval in the honeybee brain., (Copyright 2001 Academic Press.)

Disorders of taste are prevalent in patients undergoing chemotherapy and often have a negative impact on quality of life and nutrition. We now present a review of the physiology of taste and smell and a discussion of the multiple causes of disturbances in these senses in patients undergoing standard- and high-dose chemotherapy. A better understanding of the complex and often multifactorial etiology of taste dysfunction should enable the clinician to institute measures to minimize the impact of these disturbing changes.

Afrotropical malaria vectors of the Anopheles gambiae complex (Diptera: Culicidae), particularly An. gambiae sensu stricto, are attracted mainly to human hosts. A major source of human volatile emissions is sweat, from which key human-specific components are the carboxylic acids (E)- and (Z)-3-methyl-2-hexenoic acid and 7-octenoic acid. Electrophysiological studies on the antennae of An. gambiae s.s. showed selective sensitivity to these compounds, with a threshold at 10(-6) g comparable to that of known olfactory stimulants 1-octen-3-ol, p-cresol, isovaleric acid, and lower than threshold sensitivity to L-lactic acid and the synthetic mosquito repellent N,N-diethyltoluamide (DEET). A combination of the acids released at concentrations > 10(-5) g in wind tunnel bioassays significantly reduced the response to CO2, the major attractant released by human hosts, for strains of An. gambiae s.s. originating from East and West Africa. Field trials with odour-baited entry traps (OBETs) in Burkina Faso showed that 7-octenoic acid significantly increased (by 1.7-fold) the catch of females of An. gambiae sensu lato (comprising two sibling species: An. arabiensis Patton and An. gambiae s.s.) in OBETs baited with CO2, whereas combinations of the acids significantly reduced the catch in CO2-baited traps (by 2.1-fold) and in whole human odour-baited traps (by 1.5-fold). The pure (E) and (Z) geometric isomers of 3-methyl-2-hexenoic acid gave comparable results to the (EIZ) isomer mixture. These results provide the first experimental evidence that human-specific compounds affect the behaviour of highly anthropophilic An. gambiae s.l. mosquitoes. The compounds appear to inhibit the upwind flight' response to known long-range attractants, and may serve either to mask' the attractants present or, more probably, to 'arrest' upwind flight when mosquitoes arrive at a host under natural conditions. In the final approach to hosts, vectors are known to reduce their flight speed and increase their turning rate, to avoid overshooting the source. In our experimental apparatus, these changes in flight behaviour would reduce the number of mosquitoes entering the ports of the collection devices.

The biogenic amine octopamine is known to enhance the sensitivity of male moths to their species-specific pheromones in flight-tunnel experiments. This sensitization of pheromone-guided upwind flight is at least partly due to octopamine-dependent increases in the peak nerve impulse frequency of the pheromone response of olfactory receptor neurons. It is not known, however, whether octopamine exerts its effects directly on the electrical properties of the olfactory receptor neurons or indirectly, via modulation of the accessory cells of the sensillum. In extracellular tip recordings of pheromone-dependent trichoid sensilla on the antennae of male Manduca sexta moths, we investigated the effects of octopamine and serotonin on the transepithelial potential, which is generated by the activity of V-ATPases in sensillar accessory cells. In addition, the action potential activity of unstimulated olfactory receptor neurons was examined in the presence of biogenic amines. Under constant environmental conditions, the transepithelial potential oscillated regularly with periods of 2-8 min and with a 1-25mV peak-to-peak amplitude over periods of several hours. These oscillatory intervals were interrupted by periods of relatively stable transepithelial potential, correlated with flight activity by the moth. Octopamine reduced the amplitude of the transepithelial potential oscillation and decreased the resistance of the sensillum preparation in a dose-dependent manner. Serotonin altered the waveform of the transepithelial potential, but did not change the resistance of the preparation. Thus, both amines affect the accessory cells, but have different targets in the regulation of the transepithelial potential. Neither amine significantly influenced the spontaneous action potential activity of the olfactory receptor neurons.

Lake Michigan mottled sculpin, Cottus bairdi, exhibit a naturally occurring and unconditioned orienting response that can be triggered by both live prey and chemically inert vibrating spheres, even in blinded animals. CoCl(2)-induced reductions of the orienting response demonstrate that the lateral line is required for this behavior in the absence of non-mechanosensory cues (such as vision), but shed no light on the relative contributions of superficial and canal neuromasts to this behavior. To determine the relative roles of these two subsystems, we measured the frequency with which mottled sculpin oriented towards a small vibrating sphere before and after two treatments: (i) immersion of fish in a solution of gentamicin, an aminoglycoside antibiotic that damages hair cells in canal, but not superficial, neuromasts; and (ii) scraping the skin of the fish, which damages the superficial, but not the canal, neuromasts. To ensure that both superficial and canal neuromasts were adequately stimulated, we tested at different vibration frequencies (10 and 50 Hz) near or at the best frequency for each type of neuromast. At both test frequencies, response rates before treatment were greater than 70 % and were significantly greater than 'spontaneous' response frequencies measured in the absence of sphere vibration. Response rates fell to spontaneous levels after 1 day of gentamicin treatment and did not return to pre-treatment levels for 10-15 days. In contrast, response rates stayed approximately the same after superficial neuromasts had been damaged by skin abrasion. Scanning electron microscopy confirmed hair cell damage (loss of apical cilia) in canal, but not superficial, neuromasts of gentamicin-treated animals after as little as 24 h of treatment. The sensory epithelium of canal neuromasts gradually returned to normal, following a time course similar to behavioral loss and recovery of the orienting response, whereas that of superficial neuromasts appeared normal throughout the entire period. This study shows that the orienting response of the mottled sculpin is mediated by canal neuromasts.

Sawa-J, CSJ02 and N601 x C601 are selected mutant strains of Bombyx mori, which grow on various artificial diets or temporarily ingest various plant leaves. To examine the mechanisms mediating diet breadth of caterpillars, gustatory spike responses of the silkworms, called 'polyphagous" strains, were compared with normal strains, N137 x C146 and C02. There were notable differences in their feeding habits and in their sensitivity to salicin in deterrent cells in the maxillary medial styloconic sensilla and the epipharyngeal sensilla. By contrast, the deterrent cells of all strains responded similarly to strychnine nitrate in a dose-dependent manner. In additional comparisons of Sawa-J and N137 x C146, Sawa-J maxillary deterrent cells were significantly less sensitive to phloridzin, amygdalin and arbutin, but responded to some alkaloids and 20-hydroxyecdysone with similar or even higher firing rate. These results suggest that the deficiency of the sensitivity to some deterrent compounds on the deterrent cell of the polyphagous strains may be caused by mutant genes and affects the diet breadth of caterpillars.

The effects of exo- and endogenous cGMP on the resting activity (RA) of afferent crista fibers were studied in isolated preparations of the statocysts of the cuttlefish Sepia officinalis and the squid Sepioteuthis lessoniana. Bath application of the membrane-permeable cGMP analogs 8-bromo-cGMP (B-cGMP) and N(2),2'-o-dibutyryl 3', 5'-cyclic guanosine monophosphate (dB-cGMP), and of the selective inhibitor of cGMP-phosphodiesterase zaprinast (ZAP), caused an inhibition of RA. The inhibitory effects of B-cGMP and dB-cGMP remained when the preparation was pre-treated with: (i) the guanylate cyclase inhibitors 1H-[1,2,4]oxadiazolo[4,3, -a]quinoxalin-1-one (ODQ) or cystamine (CYS); (ii) the adenylate cyclase inhibitors nicotinic acid (NIC-A), 2',3'dideoxyadenosine (DDA), or MDL-12330A (MDL); (iii) the guanylate cyclase inhibitor methylene blue (M-BLU) and the adenylate cyclase inhibitor MDL combined; or (iv) the nitric oxide (NO) synthase inhibitors N(G)-nitric-L-arginine methyl ester HCl (L-NAME) or N(G)-nitro-L-arginine (L-NOARG). These data indicate that cGMP, as an intracellular messenger, has a tonic inhibitory effect on the RA of afferent crista fibers in cephalopod statocysts.