A histochemical and electrophysiological study of the action of diazoxon on cholinesterase activity and nerve conduction in ganglia of the cockroach Periplaneta americana L

When sixth abdominal or metathoracic ganglia of the cockroach Periplaneta americana L. were irrigated continuously with diazoxon (O, O-diethyl O-(2-isopropyl-6-methyl-4-pyrimidinyl) phosphate) solution in situ, the log. of the time required to block conduction in certain nerve pathways in the ganglia was directly proportional to the log. of the concentration of diazoxon applied. Inhibition of cholinesterase began peripherally before function was affected, and had begun to affect the neuropile by the time that conduction was first blocked. Longer exposure to diazoxon disrupted nerve function even more, especially in the sixth abdominal ganglion, and inhibited more cholinesterase, but much longer exposure was needed to inhibit nearly all the cholinesterase. Irrigation with saline, begun when block first occurred, failed to restore completely either nerve function or cholinesterase activity. The cholinesterase activity of ganglia from cockroaches treated topically with an LD90 of diazoxon and examined at intervals after treatment decreased steadily to a level similar to that of ganglia treated directly with diazoxon until conduction was just blocked, but rarely became less, even in moribund insects. Nerve function in metathoracic ganglia became badly affected and remained so in all cockroaches that failed to recover, but sixth abdominal ganglia, though usually badly affected for a time, always recovered normal function, even in prostrate cockroaches. The condition of a poisoned insect, therefore, corresponded much more closely to the functional condition of the metathoracic ganglion than to that of the sixth abdominal ganglion. Applying the insecticide close to a ganglion advanced the time of onset of symptoms but affected the final outcome very little. It was calculated that the highest concentration of diazoxon in the haemo-lymph in contact with the nervous systems of cockroaches treated topically with LDgo's of diazoxon was about 10^-5M.     

The mode of action of pyrethnxm on the cockroach, Periplaneta americana L

Pyrethrin, the active principle of pyrethrum powder, is insoluble in water but is soluble in, the body fluid of the cockroach. It has a selective action on nerve ganglia, and the destruction of their cells is responsible for the death of the insect. Whereas pyrethrum, when used -either in the powdered form or in a fluid state mixed with kerosene and introduced directly into the body cavity reaches the ganglion with the circulation, its mode of action when it acts through the spiracles is different. Kerosene-pyrethrum mixture when introduced into the tracheal trunks through the spiracles is quickly diffused into the haemocoele. When the dry powder is inserted into the trachea, its mode of action is analogous to a fluid preparation. As the conversion of pyrethrum from a dry into a fluid state is alone possible in the interior of the trachea, the natural conclusion is that a fluid analogous to the body fluid is present in the same situation. As soon as the pyrethrin is dissolved, it is quickly diffused into the haemocoele and thus reaches the nearest ganglion.     

Alkaline phosphatase activity in the brain of the American cockroach Periplaneta americana L

Summary The supra- and suboesophageal ganglia of the American cockroach contain material which catalyses the alkaline hydrolysis (pH 9.5) of 5-bromo-4-chloro-3-indolyl phosphate in the presence of Nitro blue tetrazolium. Histochemical studies on unfixed cryostat sections indicate that this type of alkaline phosphatase is restricted to discrete regions in the cockroach brain. Highest enzyme activity is encountered in the mushroom bodies, central body, antennal glomeruli and specific parts of some distinct neural connections including the optic nerve, antennal nerve, circumoesophageal connectives and nerves leaving the suboesophageal ganglion. Tissue fixation by use of formaldehyde-type fixatives, as well as routine paraffin-embedding, completely destroy all histochemically detectable enzyme activity.Native polyacrylamide gradient electrophoresis suggests that the alkaline phosphatase activity is present as multiple isozymic forms, which show up in the 120–130 kD range of standard proteins. Enzyme activity becomes undetectable after fixation (trichloroacetic acid, formaldehyde containing fixatives) of electrophoretically separated native proteins, as well as after electrophoresis in denaturing conditions (SDS and -mercapto-ethanol, boiling). However, the enzyme activity remains virtually unaffected after storage of the sample for prolonged periods at –20 to –80°C.     

Organization of giant interneuron projections in thoracic ganglia of the cockroach Periplaneta americana

We have investigated the structural organization of the wind-sensitive giant interneurons in the thoracic ganglia of adult American cockroaches. These seven bilaterally paired interneurons have long been thought to play a role in directing the wind-elicited escape response of the animal. Each of the giant interneurons was labeled individually by intracellular injection of cobaltic hexamine chloride. An individual giant interneuron could be reliably identified from animal to animal based on its branching pattern in thoracic ganglia. The axons of the giant interneurons are situated on each side of the nerve cord in two recognizable subgroups. Comparisons of the axonal arbors of the dorsal and ventral subgroups showed that they project into distinct but partly overlapping regions of thoracic ganglia. Three of the giant interneurons were found to have axonal arbors that cross the longitudinal midline of thoracic and abdominal ganglia. Bilateral pairs of these giant interneurons were labeled concomitantly, and many of the individual neurites from these cells appeared to be closely apposed. All these morphological characteristics are discussed in relation to the connectivity and functional significance of the giant interneurons.     

Characterization of tactile-sensitive interneurons in the abdominal ganglia of the cockroach,Periplaneta americana

Tactile stimulation of an insect's abdomen evokes various behaviors including grooming and vigorous escape responses. We tested a sample of 37 tactile-sensitive abdominal interneurons for various morphological and physiological characteristics, including their ability to excite thoracic interneurons that are known to integrate wind information conducted by giant interneurons in the classical escape response. The results suggest that abdominal tactile-sensitive interneurons are heterogeneous both in anatomical and physiological properties. In general, these cells are very small interganglionic interneurons that respond to tactile stimulation at more than one abdominal segment. However, the larger population contained virtually all types of cells. Some projected anteriorly, others posteriorly, and still others projected in both directions. For most cells, the soma was on the side opposite to their axons, but in 24% of the cells it was on the same side. Patterns of dendritic arbors also varied among cells. However, tactile sensitivity was in general consistent with the morphological bias noted in dendritic branch patterns. We were able to document the existence of tactile abdominal interneurons that connect directly to thoracic interneurons involved in escape (TI_As). However, instances of demonstrated connectivity were rare. One cell that did show connectivity (AI652) was characterized in detail, and its properties were appropriate for conducting tactile signals in a directional escape system. The dendritic arbors were biased to the side that was ipsilateral to the cell's soma and axon. As a result, this cell's abdominal inputs and thoracic outputs are on the same side. This pattern is appropriate for generating the sensory fields recorded previously in TI_As. Its axon was located in the ventral median tract, which should bring it close to the integrating region of the TI_As. © 1998 John Wiley & Sons, Inc. J Neurobiol 34: 227–241, 1998     

Contractile activation in the retractor unguis muscle of the cockroach Periplaneta americana

1. The K⁺-induced contracture consists of a phasic and a sustained component. Both were eliminated in Ca²⁺-free saline, but the sustained component recovered on the addition of Ca²⁺ to the muscle.2. Procaine mainly inhibited the phasic component. 3. Unlike the sustained component, the phasic component was inhibited by nifedipine in a concentration dependent manner.4. Divalent cations such as Mn²⁺, Co²⁺ and Ni²⁺ markedly increased the sustained component at low concentrations, but decreased it at high concentrations. The cations also modified the phasic component differentially, but to a lesser extent. High concentration abolished the phasic component.5. Ouabain markedly enhanced the sustained component.6. Caffeine contracture was a phasic type. Its duration and amplitude were augmented by pre-soaking the muscle in Na⁺-reduced salines. Immediate pre-treatment with caffeine eliminated the phasic component of the 160mM K⁺-induced contracture.7. These results suggest that a Na-Ca exchange mechanism may play a role in excitation-contraction coupling in insect muscle. Calcium ions flowing into the cell upon membrane depolarization may specifically activate the phasic component by way of a calcium-induced calcium releasing mechanism.     

Intestinal microbial flora of the of the American cockroach, Periplaneta americana L.

A morphological study employing scanning and transmission electron microscopy was made by the alimentary tract of the American cockroach, Periplaneta americana L. A. complex microbiota of diverse morphology, which could not be readily dislodged, was observed and found to be restricted to the hindgut, particularly the colon. Numerous filamentous forms were noted, and some are described, including the morphologically distinct Methanospirillum. Flora was noted attached to the cuticular lining and cuticular filaments of the colon, and several spiral forms were observed in the luminal contents from the colon.     

The action of tetrodotoxin on the cockroach, Periplaneta americana: a toxicological and neurophysiological study

ABSTRACT. Some sublethal effects of tetrodotoxin (TTX) on the cockroach were investigated using chronically implanted extracellular electrodes. Prostration and subsequent paralysis were correlated with blockage of peripheral nerves, the CNS being apparently unaffected at the dose employed. Over several days following treatment many cockroaches showed a reversal of the symptoms and finally recovered fully. This coincided with recovery of the blocked nerves. It is suggested that TTX can be used to distinguish pharmacologically the peripheral and central actions of drugs and insecticides. Furthermore, TTX treatment could also be useful in the analysis of the subcellular nature of the sodium channels in nerve membrane.     

Tyrosine hydroxylase in the cerebral ganglia of the American cockroach (Periplaneta americana L.): an immunohistochemical study

We have investigated the distribution of tyrosine-hydroxylase-like immunoreactivity in the cerebral ganglia of the American cockroach, Periplaneta americana. Groups of tyrosine-hydroxylase-immunoreactive cell bodies occur in various parts of the three regions of the cerebral ganglia. In the protocerebrum, single large neurons or small groups of neurons are located in the lateral neuropil, adjacent to the calyces, and in the dorsal portion of the pars intercerebralis. Small scattered cell bodies are found in the outer layers of the optic lobe, and clusters of larger cell bodies can be found in the deutocerebrum, medial and lateral to the antennal glomeruli. Thick bundles of tyrosine-hydroxylase-positive nerve fibers traverse the neuropil in the proto- and deutocerebrum and innervate the glomerular and the nonglomerular neuropil with fine varicose terminals. Dense terminal patterns are present in the medulla and lobula of the optic lobe, the pars intercerebralis, the medial tritocerebrum, and the area surrounding the antennal glomeruli, the central body and the mushroom bodies. The pattern of tyrosine-hydroxylase-like immunoreactivity is similar to that previously described for catecholaminergic neurons, but it is distinctly different from the distribution of histaminergic and serotonergic neurons.     

Biosynthesis of 3-methylpentacosane in the cockroach Periplaneta americana.

The biosynthesis of 3-methylalkanes was investigated in the cockroach $\text{Periplaneta americana}$. Between 0.2 and 0.3 percent of the labelled acetate and propionate injected into the insect was incorporated into the cuticular hydrocarbons, compared to 0.01 percent for labelled isoleucine. Twenty-three ± four percent of the [2-¹⁴C]acetate, 42 ± 3 and 44 ± 4 percent of the [2-¹⁴C] and [3-¹⁴C]propionate, and 75 ± 5 percent of the [1-¹⁴C]propionate incorporated into the cuticular hydrocarbons was found in 3-methylpentacosane. These results indicate that propionate serves as the source of the branching methyl group, suggesting a pathway in which this precursor is incorporated during the penultimate step in 3-methylalkane biosynthesis in insects.     

Nicotinic acetylcholine receptors on a cholinergic nerve terminal in the cockroach,Periplaneta americana

Summary Intracellular microelectrode recording and ionophoretic application of carbamylcholine (CCh) were used to compare the cholinergic sensitivity of postsynaptic dendrites of an identified neurone with that of an identified presynaptic cholinergic axon.The axon of the lateral filiform hair sensory neurone (LFHSN) in the first-instar cockroachPeriplaneta americana was found to be as sensitive to CCh as the dendritic regions of giant interneurone 3 (GI 3). The CCh response of both neurones was unaffected by replacing Ca²⁺ with Mg²⁺, confirming that the ACh receptors are present on the neurones under test. The CCh response of both neurones was mimicked by ionophoretic application of nicotine. The responses were blocked by 10^–5 M mecamylamine and 10^–6 M d-tubocurarine and were not affected by muscarinic antagonists, suggesting that the ACh receptors present on GI 3 and LFHSN are predominantly nicotinic.The muscarinic agonist oxotremorine and the antagonists atropine and quinuclidinyl benzilate had no modulatory effect on LFHSN-GI 3 synaptic transmission.The latency of the LFHSN response to CCh was consistent with the hypothesis that ACh receptors are situated on the main axon/terminal within the neuropil of the ganglion. It has previously been shown that this region of the axon does not form output synapses (Blagburn et al. 1985a). This indirect evidence indicates that presynaptic or extrasynaptic ACh receptors are present in the membrane of a cholinergic axon.LFHSN was depolarized by synaptically-released ACh after normal or evoked spike bursts, suggesting that the nicotinic ACh receptors act as autoreceptors. However, it was not possible to obtain direct evidence to support the hypothesis that these receptors modulate ACh release.Abbreviations CCh carbamylcholine - GI giant interneurone - FHSN filiform hair sensory neurone - LFHSN lateral filiform hair sensory neurone - R _in input resistance - V depolarization - V _m resting potential