Dopamine stimulates salivary duct cells in the cockroach Periplaneta americana.
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This study examines whether the salivary duct cells of the cockroach Periplaneta americana can be stimulated by the neurotransmitters dopamine and serotonin. We have carried out digital Ca2+-imaging experiments using the Ca2+-sensitive dye fura-2 and conventional intracellular recordings from isolated salivary glands. Dopamine evokes a slow, almost tonic, and reversible dose-dependent elevation in [Ca2+]i in the duct cells. Upon stimulation with 10(-)6 mol l-1 dopamine, [Ca2+]i rises from 48+/-4 nmol l-1 to 311+/-43 nmol l-1 (mean +/- s.e.m., N=18) within 200-300 s. The dopamine-induced elevation in [Ca2+]i is absent in Ca2+-free saline and is blocked by 10(-)4 mol l-1 La3+, indicating that dopamine induces an influx of Ca2+ across the basolateral membrane of the duct cells. Stimulation with 10(-)6 mol l-1 dopamine causes the basolateral membrane to depolarize from -67+/-1 to -41+/-2 mV (N=10). This depolarization is also blocked by La3+ and is abolished when Na+ in the bath solution is reduced to 10 mmol l-1. Serotonin affects neither [Ca2+]i nor the basolateral membrane potential of the duct cells. These data indicate that the neurotransmitter dopamine, which has previously been shown to stimulate fluid secretion from the glands, also stimulates the salivary duct cells, suggesting that dopamine controls their most probable function, the modification of primary saliva. (+info)
Active signaling of leg loading and unloading in the cockroach.
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The ability to detect changes in load is important for effective use of a leg in posture and locomotion. While a number of limb receptors have been shown to encode increases in load, few afferents have been demonstrated to signal leg unloading, which occurs cyclically during walking and is indicative of slipping or perturbations. We applied mechanical forces to the cockroach leg at controlled rates and recorded activities of the tibial group of campaniform sensilla, mechanoreceptors that encode forces through the strains they produce in the exoskeleton. Discrete responses were elicited from the group to decreasing as well as increasing levels of leg loading. Discharges of individual afferents depended on the direction of force application, and unit responses were correlated morphologically with the orientation of the receptor's cuticular cap. No units responded bidirectionally. Although discharges to decreasing levels of load were phasic, we found that these bursts could effectively encode the rate of force decreases. These discharges may be important in indicating leg unloading in the step cycle during walking and could rapidly signal force decreases during perturbations or loss of ground support. (+info)
Molecular characterization of American cockroach tropomyosin (Periplaneta americana allergen 7), a cross-reactive allergen.
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Inhalation of allergens produced by the American cockroach (Periplaneta americana) induces IgE Ab production and the development of asthma in genetically predisposed individuals. The cloning and expression in Escherichia coli of P. americana tropomyosin allergen have been achieved. The protein shares high homology with other arthropod tropomyosins (80% identity) but less homology with vertebrate ones (50% identity). The recombinant allergen was produced in E. coli as a nonfusion protein with a yield of 9 mg/l of bacterial culture. Both natural and recombinant tropomyosins were purified by isoelectric precipitation. P. americana allergen 1 (Per a 1) and Per a 7 (tropomyosin) are to date the only cross-reacting allergens found in cockroaches. ELISA and Western blot inhibition experiments, using natural and recombinant purified tropomyosins from shrimp and cockroach, showed that tropomyosin induced cross-reactivity of IgE from patients allergic to these allergens, suggesting that this molecule could be a common allergen among invertebrates. (+info)
Chemo-anemotaxis: a behavioral response to sex pheromone in nonflying insects.
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The cockroach, Periplaneta americana, running on a Y-ring globe, moves downwind if an air current is directed toward the head. However, if the air current carries sex pheromone, then upwind movement is elicited. This orientation behavior is apparently a mechanism to facilitate the orientation of males searching for pheromone-secreting females. (+info)
Double-stranded RNA interference shows that Engrailed controls the synaptic specificity of identified sensory neurons.
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The transcription factor Engrailed (En) controls the topography of axonal projections by regulating the expression of cell-adhesion molecules [1] [2] [3] [4] but it is not known whether it also controls the choice of individual synaptic target cells. In the cercal sensory system of the larval cockroach (Periplaneta americana), small numbers of identified wind-sensitive sensory neurons form highly specific synaptic connections with 14 identified giant interneurons [5] [6], and target-cell choice is independent of the pattern of axonal projections [6]. En is a putative positional determinant in the array of cercal sensory neurons [7]. In the present study, double-stranded RNA (dsRNA) interference [8] was used to abolish En expression. This treatment changed the axonal arborisation and synaptic outputs of an identified En-positive sensory neuron so that it came to resemble a nearby En-negative cell, which was itself unaffected. We thus demonstrate directly that En controls synaptic choice, as well as axon projections. (+info)
Preparation of (-)-periplanone D and its physical and spectroscopic properties.
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(-)-Periplanones C and D were obtained in reproducible yields by modifying reported procedures. Our synthetic sample of (-)-periplanone D showed somewhat different physical and spectroscopic properties from those reported in the literature. (+info)
Peptide-induced Ca(2+) movements in a tonic insect muscle: effects of proctolin and periviscerokinin-2.
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Although most of the characterized insect neuropeptides have been detected by their actions on muscle contractions, not much is known about the mechanisms underlying excitation-contraction coupling. Thus we initiated a pharmacological study on the myotropic action of the peptides periviscerokinin-2 (PVK-2) and proctolin on the hyperneural muscle of the cockroach Periplaneta americana. Both peptides required extracellular Ca(2+) to induce muscle contraction, and a blockage of sarcolemmal Ca(2+) channels by Mn(2+) or La(3+) inhibited myotropic effects. The peptides were able to induce contractions in dependence on the extracellular Ca(2+) concentration in muscles depolarized with high K(+) saline. A reduction of extracellular Na(+), K(+), or Cl(-) did not effect peptide action. Nifedipine, an L-type Ca(2+)-channel blocker, partially blocked the response to both peptides but to a much lesser extent than contractions evoked by elevated K(+). Using calcium imaging with fluo-3, we show that proctolin induces an increase of the intracellular Ca(2+) concentration. In calcium-free saline, no increase of the intracellular Ca(2+) concentration could be detected. The inhibiting effect of ryanodine, thapsigargin, and TMB-8 on peptide-induced contractions suggests that Ca(2+) release from the sarcoplasmic reticulum plays a major role during peptide-induced contractions. Preliminary experiments suggest that the peptides do not employ cyclic nucleotides as second messengers, but may activate protein kinase C. Our results indicate that the peptides induce Ca(2+) influx by an activation or modulation of dihydropyridine-sensitive and voltage-independent sarcolemmal Ca(2+) channels. Ca(2+)-induced Ca(2+) release from intracellular stores, but not inositol trisphosphate-induced Ca(2+) release, seems to account for most of the observed increase in intracellular Ca(2+). Additionally, both peptides were able to potentiate glutamate-induced contractions at threshold concentrations. (+info)
Peptidergic modulation of an insect Na(+) current: role of protein kinase A and protein kinase C.
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The modulation of voltage-gated Na(+) currents in isolated somata of dorsal unpaired median (DUM) neurons of the cockroach Periplaneta americana was investigated using the patch-clamp technique. The neuropeptide Neurohormone D (NHD), which belongs to the family of adipokinetic hormones, reversibly reduced the Na(+) current in concentration-dependent manner (1 pM to 10 nM). At 10 nM, NHD caused an attenuation of the maximum of current-voltage (I-V) relation for peak currents by 23 +/- 6%. An analysis of NHD action on current kinetics in terms of the Hodgkin-Huxley formalism revealed that NHD reduces the time constant of inactivation, whereas steady-state activation and inactivation as well as the time constant of activation were not affected. In addition, NHD prolonged the recovery from inactivation. The cAMP analogue 8-bromo-cAMP, forskolin, and the catalytic subunit of protein kinase A mimicked the action of NHD. Furthermore, preincubation of cells with the protein kinase A inhibitor KT 5720 abolished the action of NHD. Thus NHD seems to modify the Na(+) current via channel phosphorylation by protein kinase A. Activation of protein kinase C by oleoylacetylglycerol (OAG) also reduced the Na(+) current, but it did not occlude the action of NHD. On the other hand, inhibition of protein kinase C by chelerythrine or Go 6976 did not essentially impair the NHD effects. (+info)