Organization of motoneurones in the prothoracic ganglion of the cockroach Periplaneta americana (L.).

The location within the prothoracic ganglion of neurone somata with axons in identified peripheral nerves is examined by the cobalt iontophoresis technique. Axons are filled with cobalt by diffusion through their cut ends and the cobalt is then precipitated as the black sulphide inside the neurone. It is assumed that neurones with axons in peripheral nerves and somata in central ganglia are either motor or neuro-secretory. Fifteen nerves are examined and maps of the location of somata with axons in each nerve are presented. The axon distribution in peripheral nerves of three common inhibitory neurones is described. Dendritic morphology of one common inhibitory neurone and two coxal depressor motoneurones is illustrated. It is proposed that some individual neurones can be reliably identified from their soma dimensions and location within the ganglion. The number of motoneurones with somata in the prothoracic ganglion and their homology with cells in the other thoracic ganglia are discussed.     

Localization of corazonin in the nervous system of the cockroach Periplaneta americana

Antisera raised to the cardioactive peptide corazonin were used to localize immunoreactive cells in the nervous system of the American cockroach. Sera obtained after the seventh booster injection were sufficiently specific to be used for immunocytology. They recognized a subset of 10 lateral neurosecretory cells in the protocerebrum that project to, and arborize and terminate in the ipsilateral corpus cardiacum. They also reacted with bilateral neurons in each of the thoracic and abdominal neuromeres, a single dorsal unpaired median neuron in the suboesophageal ganglion, an interneuron in each optic lobe, and other neurons at the base of the optic lobe, in the tritocerebrum and deutocerebrum. The presence of corazonin in the abdominal neurons and the lateral neurosecretory cells was confirmed by HPLC fractionation of extracts of the abdominal ganglia, brains and retrocerebral complexes, followed by determination of corazonin by ELISA, which revealed in each tissue a single immunoreactive peak co-eluting with corazonin in two different HPLC systems. Antisera obtained after the first three booster injections recognized a large number of neuroendocrine cells and neurons in the brain and the abdominal nerve cord. However, the sera from the two rabbits reacted largely with different cells, indicating that the majority of this immunoreactivity was due to cross-reactivity. These results indicate that the production of highly specific antisera to some neuropeptides may require a considerable number of booster injections.     

Submicroscopic characterization of proctolin-like immunoreactivity in the nervous system of the cockroach Periplaneta americana L.

Summary Proctolin-like immunoreactivity (PLI) was found in different parts of the arthropod central nervous system and in nerve fibers of muscles. In order to examine whether this PLI is related to a uniform type of secretory vesicle, hindgut musculature and frontal and hypocerebral ganglia were examined with the immunogold technique. PLI occurs exclusively within membran-bounded secretory granules. Neither granular ER nor Golgi stacks show PLI. In some cases close relationships between PLI-bearing granules and lysosomes were observed. In presynaptic areas, PLI-reactive granules are associated with numerous clear synaptic vesicles and restricted to an area distinctly separate from the presynaptic membrane. Three types of granules were found, differing in diameter and electron density: (1) dense, 80 nm; (2) dense, 150 nm; (3) low density, 150 nm. The results demonstrate that: (1) the PLI of the produced peptide occurs shortly after its separation from the Golgi stack; (2) the occurrence of PLI in three different granule types could be the morphological expression of the common occurrence of proctolin with other neuroactive substances. However, a possible cross-reactivity with other, hitherto unknown substances must be considered as well.     

The uptake of L-glutamate by the central nervous system of the cockroach, Periplaneta americana.

A concentrative uptake mechanism for L-glutamate with the following characteristics has been identified in the abdominal nerve cord: 1. The uptake can be divided into Na+-sensitive and Na-plus-insensitive components. 2. The Na-plus-sensitive component showed the typical saturation kinetics of a carrier mediate process. It had a V of 15.9 x 10(6) times 10(6) muM/mg wet weight/min and a Km of 0-33 mm. Its magnitude was proportional to the first power of the Na-plus concentration of the medium. The uptake was specific for L-dicarboxylic amino acids and was sensitive to the presence of metabolic inhibitors. 3. The Na-plus-insensitive component was linearly related to the glutamate concentration of the medium. An isosmotic saline is described for use with the isolated intact abdominal nerve cord of P. americana.     

Serotonin-immunoreactive visceral nerves and neurohemal system in the cockroach Periplaneta americana (L.)

Summary By use of the indirect immunofluorescent technique applied to whole mounts of tissues and organs of the American cockroach, Periplaneta americana, serotonin-like reactivity has been demonstrated in an extensive meshwork of fibers on the surface of nerves of the subesophageal ganglion and the tritocerebrum. This meshwork appears to serve the neurohemal release of serotonin. In addition, several of these nerves contain two or more centrally located axons that appear to be serotonergic. The corpora cardica show immunoreactivity but do not appear to be a principal release site for serotonin. The nerves and ganglia of the stomadeal nervous system show immunoreactivity for serotonin and provide serotonin-positive innervation to the salivary glands and the visceral muscles of the stomodeum and midgut. The heart, hindgut, Malpighian tubules, fat body, and skeletal muscles all appear to lack serotonin immunoreactivity.     

Actions of deltamethrin and tralomethrin on cholinergic synaptic transmission in the central nervous system of the cockroach (Periplaneta americana)

Deltamethrin (RU 22974) and tralomethrin isomers (RU 24784 and RU 24785) block transmission at the cercal-afferent giant-interneuron synapses of the cockroach, when bath-applied to the desheathed ganglion at micromolar concentrations. The time-course of the events leading to the blockage suggests two possible target sites: one located presynaptically and the other situated on postsynaptic membranes.     

[N-Methyl-3H]Scopolamine binding sites in the central nervous system of the cockroach Periplaneta americana

The binding of [N-methyl-³H]scopolamine to a cockroach nerve cord preparation has been investigated. Specific [N-methyl-³H]scopolamine binding was found to be saturable and of high affinity (K_d = 13.9 nM). Muscarinic ligands were found to displace [N-methyl-³H]scopolamine binding more effectively than nicotinic ligands. The distribution of these [N-methyl-³H]scopolamine binding sites was examined in the metathoracic ganglion at the light microscope level by autoradiographical techniques. Specific binding was found to be localized to distinct regions of the neuropile. This pattern showed certain similarities to that seen when the ganglion was stained for acetylcholinesterase, suggesting a functional role for these insect muscarinic acetylcholine receptors.     

Cercal sensory regulation of acetylcholinesterase in nervous system of the cockroach, Periplaneta americana

Cercal ablation caused a significant loss in acetylcholinesterase (AChE) activity of the cercal nerves and terminal ganglion within 12 hr while a similar reduction in enzyme activity of connectives was noticed at least one day after cercectomy. The decrease in AChE activity of the nervous tissues showed a recovery toward control levels from 20 days of unilateral cercectomy whereas the bilateral cercectomy produced a continuous and irreversible decline in enzyme activity. These localized changes in AChE activity of the abdominal nervous system of the cockroach were attributed to be regulated by the cercal sensory innervation.     

Identification of neuronal pathways between the stomatogastric nervous system and the retrocerebral complex of the cockroach Periplaneta americana (L.)

Summary The neuronal pathways connecting the stomatogastric nervous system with the retrocerebral complex of the cockroach, Periplaneta americana, were investigated by means of axonal cobalt chloride iontophoresis. Somata in the hypocerebral ganglion and in the nervus recurrens sending their axons to different parts of the stomatogastric nervous system were traced. Some axons in the oesophageal nerve arise from large perikarya in the anterior part of the pars intercerebralis and pass via the NCCI to the corpora cardiaca and the oesophageal nerve. They form a profuse dendritic tree in the protocerebrum. Fibers of the NCC I and NCC II as well as the NCA I and NCA II enter the stomatogastric nervous system via the hypocerebral ganglion.     

The spread of topically-applied pyrethrin I from the cuticle to the central nervous system of the cockroach Periplaneta americana

Penetration of topically-applied pyrethrin I into adult male American cockroaches (Periplaneta americana L.) and its subsequent distribution within the insects was studied microchemically and biologically. After applying an LD95 (0.5 g per insect), pyrethrin I was lost from the surface of the insects at a rate diminishing with time; 20% of the dose applied penetrated during the first hour, but elimination limited the amount found inside the insects to a maximum of 13%. The amount inside the insects increased for an hour after dosing, when symptoms of poisoning had become severe, but then remained steady until the end of the test, 48 hrs after dosing.Pyrethrin I sorbs strongly but reversibly on insect solids from aqueous solution, and at equilibrium is distributed between solids and solution in the ratio 3 × 10⁴: 1. This ratio would give a concentration of only 4 × 10^–11 M of pyrethrin I in the haemolymph of insects poisoned with an LD95 of the insecticide, but solids in the haemolymph would increase its pyrethrin I content and speed the spread of the insecticide from cuticle to nervous system. Chemical tests able to detect pyrethrin I at concentrations of 2×10^–8 M failed to show its presence in the haemolymph of poisoned cockroaches, but the haemolymph caused symptoms resembling those of pyrethroid poisoning when applied to nerve preparations from normal cockroaches.Although pyrethrin I at 4 × 10^–11 M in the haemolymph of cockroaches treated topically with LD95s of the insecticide seems too dilute to have produced the symptoms observed in their nervous systems, the results of additional tests did not support the alternative suggestion that the insecticide reached the nervous system by spreading over the cuticle and through the tracheal system. Larger concentrations of pyrethrin I occurring locally in the haemolymph near the nervous system during the early stages of poisoning may explain the anomaly.

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Résumé La pénétration d'une application topique de pyrethrine I chez le mâle adulte de Periplaneta americana et sa distribution ultérieure dans l'organisme est étudiée microchimiquement et biologiquement. Après application d'une dose égale à la DL 95 (0,5 g par insecte), la pyrethrine I est absorbée à la surface de l'insecte à un taux décroissant avec le temps; 20% de la dose appliquée pénètre durant la 1ère heure, mais l'élimination limite la quantité trouvée dans l'organisme à un maximum de 13%. La quantité trouvée dans l'organisme s'accroît pendant la 1ère heure après l'intoxication, quand les symptômes de l'intoxication deviennent importants, puis reste stationnaire jusqu'à la fin du test, soit 48 heures après l'intoxication.La pyrethrine I s'absorbe fortement mais réversiblement sur la phase solide de l'insecte, à partir de la solution aqueuse et le coefficient de partition est de 3×10⁴: 1. Ce coefficient donnarait une concentration de pyrethrine I de seulement 4 × 10^–11 M dans l'hémolymphe des insectes intoxiqués à la DL 95, mais la phase solide de l'hémolymphe augmenterait sa teneur en pyrethrine I et accélérerait la vitesse de pénétration de l'insecticide, de la cuticule vers le système nerveux. Des tests chimiques capables de déceler la pyrethrine I à la concentration de 2×10^–8 M n'ont pu établir sa présence dans l'hémolymphe des blattes intoxiquées, mais cette hémolymphe provoque des symptômes analogues à ceux de l'intoxication par les pyrethrines, quand elle est appliquée à des préparations nerveuses de blattes nonintoxiquées.Quoique la pyrethrine I à la concentration de 4 × 10^–11 M dans l'hémolymphe de blattes traitées localement à la DL 95 de cet insecticide, semble trop diluée pour produire les symptômes observés dans le système nerveux, des résultats supplémentaires n'ont pas autorisé une autre hypothèse, à savoir que l'insecticide atteindrait le système nerveux via la cuticule et le système trachéen. Des concentrations plus fortes de pyrethrine I, localisées près du système nerveux pendant les premiers stades de l'intoxication pourraient expliquer cette anomalie.

     

美洲大蠊中枢DUM神经元的分离和电压门控Na+电流的记录

【目的】建立美洲大蠊Periplaneta americana中枢神经系统背侧不成对中间神经元（dorsal unpaired median neurons, DUM neurons）的分离方法和DUM神经元电生理实验模型。【方法】IA型胶原酶法消化美洲大蠊末端腹神经节, 机械吹打得到DUM神经元细胞, 运用膜片钳技术记录DUM神经元细胞电压门控Na+电流。【结果】分离得到的DUM神经元细胞状态良好, 具有DUN神经元典型的梨状形态和表面特征。以膜片钳全细胞方式记录到的Na+电流符合钠通道电流特征。【结论】IA型胶原酶消化得到美洲大蠊DUM神经元细胞的方法可靠, 能稳定地记录到Na+电流。本文描述的方法为昆虫神经细胞的电生理机制研究提供一个可用的实验模型。