Fine structure of the first optic ganglion (lamina) of the cockroach, Periplaneta americana

The structural organization of the first optic ganglion (lamina) of the cockroach (Periplaneta americana) was investigated by the use of light and electron microscopy. Each compound eye of the cockroach is composed of up to 2000 visual units (ommatidia) of the fused rhabdom type. The ommatidia themselves consist of eight receptor cells which terminate as axons in either the first or second optic ganglion. Three different short visual fibre types end in two separate strata in the lamina, and one long fibre type ends in the second optic ganglion. Monopolar second-order neurons with wide field branching patterns in the middle stratum of the first synaptic region have postsynaptic contacts with short visual fibres. Horizontal fibre elements with branching patterns at different levels of the lamina apparently form three horizontal plexuses with presynaptic and/or postsynaptic connections to first- and secondorder neurons. The lack of well-organized fibre cartridges containing a constant number of first and second order neurons in each fascicle and the presence of only unistratified wide field monopolar cells could represent, as compared to other insect orders, a primitive stage in the development of the first optic ganglion.     

One trial learning in the American cockroach, Periplaneta americana

A new and simple method of training intact cockroaches was designed. (1) Cockroaches were maintained before and during the experiment with a high motivation to look for sugar. They were kept in a chamber with water and yeast extract ad libitum but without any sugar. (2) Cockroaches were individually trained to associate two artificial scent signals, i.e. menthol and vanilla, with sugar and salt solutions. Discriminatory learning performance was measured by testing the scent preference before and after training. Care was taken to exclude other than olfactory cues in the discriminatory paradigm. (3) Cockroaches exhibited a spontaneous initial preference for vanilla over menthol. This preference could be modified by training. The insect showed fast learning and long retention. One training trial was enough to reverse the initial preference with a significant retention after 7 days. Reversal of trained preference was accomplished by retraining. (4) The method provides an opportunity to study sensory performance and memory consolidation in cockroaches.     

Haemolymph lipase activity in the American cockroach, Periplaneta americana

The haemolymph of the cockroach, Periplaneta americana, has been shown to possess pronounced lipolytic activity. The enzyme may be considered a ‘true’ lipase because of its ability to hydrolyse long chain fatty acid glycerides. The enzyme exhibits dual optima at pH 4·9 and 7·0 and is inhibited by fluoride ion. Lipase activity in the intact cockroach is unaffected by the corpus cardiacum hypolipemic factor; however, prolonged periods of starvation result in a significant decline in enzyme activity.     

Quantitative analysis of male responses released by female sex pheromone in Periplaneta Americana

Serial dilutions of purified female extracts of sex pheromone were analysed for their activity in releasing courting behaviour in males. Running activity and courting displays by males increased with the higher concentrations of sex pheromone. Males responded to sex pheromone in three distinct levels: arousal, running and courting displays. Individual male responses to equal dilutions were variable in that certain males had higher thresholds and required higher concentrations to initiate similar responses. Each level, respectively, was initiated by increasing concentrations of sex pheromone. When exposed to the highest concentration, there was a minimal temporal response time prior to the terminal courting displays. Secondarily purified extracts, as low as 4·5×10^?14 g, elicited a response from males.     

An ultrastructural study and stereological analysis of the colon wall in the cockroach Periplaneta americana

The colonic epithelium has been examined for ultrastructural evidence of physiological activity. The cells show extensive folding of the apical plasma membrane, associated with mitochondria and an internal coating of particles about 120 Å diameter. Anteriorly many apical infoldings are dilated at the tip to form substantial extracellular spaces up to 0.8 μm wide. Narrow intercellular channels are present, opening to the haemolymph side of the epithelium. Pinocytosis is seen frequently at the basal surface. The surface densities of apical plasma membranes were not significantly different in the posterior mid-gut and colon. Similarly the volume densities of mitochondria were equal in the colonic epithelium and rectal pads, but the surface density of outer mitochondrial membranes was greater in the colon. It is suggested that the colon may absorb organic solutes from the gut lumen.     

Circadian rhythmicity in the nervous system of the cockroach, Periplaneta americana

Diurnal rhythmicity of nervous activity in Periplaneta americana was investigated, using acetylcholine (ACh) content, acetylcholinesterase (AChE) and spontaneous electrical activities as indices. AChE and electrical activities were maximum at 0 hr and minimum at 12 hr, while ACh showed an opposite rhythm. Central nervous system extract from cockroaches at 12 hr elevated the electrical activity while 0 hr-extract exerted inhibition. Lower concentrations of ACh had an elevatory influence while higher concentrations inhibited the electrical activity. A hypothesis is proposed, suggesting synthetic and releasing phases of ACh in a regular diurnal cycle, to explain the results obtained.     

Serotonin in the central nervous system of the cockroach, Periplaneta americana

Studies on serotonin in the insect nervous system has long been neglected, although serotonin is a putative neurotransmitter. During the course of this study the serotonin content was found to be significantly higher than that found in mammalian midbrain. Parachlorophenylalanine was found to inhibit the first step of the biosynthetic pathway by inhibiting tryptophan-hydroxylase enzyme and leading to alterations in the concentrations of metabolites such as 5-hydroxy tryptophan, 5-hydroxy indole acetic acid and tryptophan. Using a dose of 15 μg/g the inhibitory effect was not long lasting and recovery was observed to restore the normal levels. Higher trytophan levels were observed after a certain period of P-chlorophenylalanine treatment because there was a block in the biosynthetic path and tryptophan could not be utilized for 5-HT synthesis. A negative correlation between brain tryptophan and protein content was observed in both the cases of P-chlorophenylalanine and reserpine treatments.     

Autolysis in axon terminals of a new neurohaemal organ in the cockroach Periplaneta americana

An ultrastructural investigation showed that there was a neurohaemal organ in the wall of the ampulla of the antennal pulsatile organ. The neurosecretory axon terminals occurred singly or in small groups rather than closely packed together as in other neurohaemal organs. All axons contained the same type of neurosecretory granule. The granules had varying electron density and a diameter in the range 1000–2500 Å. Some terminals contained small, elliptical, electron-transparent vesicles and the axolemma was apposed to the stroma. Other terminals were large and enveloped by glial tissue and the contents of the terminals exhibited varying degrees of autolytic degeneration. Autolysis was characterized by the occurrence of dense bodies and multilaminate bodies which enclosed mitochondria and neurosecretory granules. It was suggested that the neurosecretory material affects antennal function.     

α-Glucosidase activity in haemolymph of the American cockroach, Periplaneta americana

α-Glucosidase activity of whole haemolymph has been investigated in adult males of the American cockroach, Periplaneta americana. Two electrophoretically distinguishable enzymes capable of hydrolysing α-glucosidic linkages are present in the serum component of the haemolymph, and one of these hydrolyses trehalose. Trehalase activity is also present in haemocytes, and the haemocyte enzyme shares an identical electrophoretic mobility and similar pH sensitivity with the serum trehalase. Furthermore, both enzymes are inhibited to the same extent by sodium ethylene diamine tetracetate (EDTA); thus it is suggested that the same enzyme may be responsible for trehalase activity in the two components. The K_m of EDTA-inhibited trehalase is 3·3 mM and this value is reduced to 1·8 mM upon activation of the enzyme by calcium ions. The properties of the trehalase are discussed in light of the possible rôle of the enzyme in regulating haemolymph trehalose and glucose concentrations.     

Fate of glucose in haemolymph of the American cockroach, Periplaneta americana

The rate of removal of high concentrations of glucose (10 μg/μl haemolymph) from haemolymph of adult male cockroaches, Periplaneta americana, was studied in normal and ligated insects. More than 50% of the injected glucose is removed from the haemolymph of normal insects within 20 min of injection. A period of rapid trehalose synthesis occurs during the initial 10 min following injection of glucose into the haemocoele, and this is succeeded by a period of glycogen synthesis. The results are discussed in terms of earlier observations on ‘stress-induced hypertrehalosemia’ and the possible involvement of a glycogenic agent.     

Functional analysis of Scr during embryonic and post-embryonic development in the cockroach, Periplaneta americana

The cockroach, Periplaneta americana represents a basal insect lineage that undergoes the ancestral hemimetabolous mode of development. Here, we examine the embryonic and post-embryonic functions of the hox gene Scr in Periplaneta as a way of better understanding the roles of this gene in the evolution of insect body plans. During embryogenesis, Scr function is strictly limited to the head with no role in the prothorax. This indicates that the ancestral embryonic function of Scr was likely restricted to the head, and that the posterior expansion of expression in the T1 legs may have preceded any apparent gain of function during evolution. In addition, Scr plays a pivotal role in the formation of the dorsal ridge, a structure that separates the head and thorax in all insects. This is evidenced by the presence of a supernumerary segment that occurs between the labial and T1 segments of RNAiScr first nymphs and is attributed to an alteration in engrailed (en) expression. The fact that similar Scr phenotypes are observed in Tribolium but not in Drosophila or Oncopeltus reveals the presence of lineage-specific variation in the genetic architecture that controls the formation of the dorsal ridge. In direct contrast to the embryonic roles, Scr has no function in the head region during post-embryogenesis in Periplaneta, and instead, strictly acts to provide identity to the T1 segment. Furthermore, the strongest Periplaneta RNAiScr phenotypes develop ectopic wing-like tissue that originates from the posterior region of the prothoracic segment. This finding provides a novel insight into the current debate on the morphological origin of insect wings.     

Water and ion balance in the tissues of the dehydrated cockroach, Periplaneta americana

Cockroaches dehydrated for 8 days lost nearly 50% of their haemolymph volume and approx 25% of their tissue water. Haemolymph osmolality and sodium, potassium, and chloride concentrations in the haemolymph and tissue water were all regulated within narrow limits. It is confirmed that sodium and potassium ions are sequestered within the fat body during periods of dehydration. The increase in sodium and potassium ions in the fat body is shown to arise from ionic regulation of haemolymph and other tissues. During periods of rehydration, sodium and potassium concentrations decrease in the fat body and haemolymph volume and ionic concentrations return to near original levels. A small proportion of the surplus haemolymph chloride ions is shown to be associated with the cuticle during times of water deprivation.     

Locomotory activity in the extensor and flexor tibiae of the cockroach, Periplaneta americana

Electromyograms (EMGs) were recorded from the metathoracic extensor and flexor tibiae of cockroaches when the animals were: walking on a level surface, walking on a ball, or producing rhythmic leg movements while being restrained ventral surface upward. In the rapidly walking (> 2 steps/s) and restrained animals, there was reciprocity between EMGs from the extensor and flexor tibiae. In slowly walking (<-2 steps/s) animals there was a conspicuous overlap in the flexor and extensor EMGs. The overlap was due to an increase in duration in the activity of the flexor. In experiments in which distal portions of a limb were amputated, the overlap observed in slow walking was either reduced greatly or lost entirely. These results are in agreement with recent locomotory models which state that the motor output is produced by a central pattern generator but can be modified by peripheral sensory inputs.     

Autoradiographic localization of [3H]-GABA accumulation in the sixth abdominal ganglion of the cockroach, Periplaneta americana L.

Localization of [³H]-GABA accumulation in the sixth abdominal ganglion of the cockroach was achieved using autoradiography.Light and electron microscope autoradiographic results provide direct evidence for glial accumulation of [³H]-GABA via a Na⁺-dependent carrier system. The functional role of the glial uptake of GABA is discussed.     

Presynaptic inhibition in the cercal-afferent giant-interneurone synapses of the cockroach, Periplaneta americana L.

The occurrence of presynaptic control of synaptic transmission in the cercal-afferent giant-interneurone system of the cockroach was investigated. Reduction in amplitude (up to 50%, lasting about 200–250 ms) of the compound evoked EPSP followed repetitive (300 to 500 Hz) supra-threshold stimulation of cercal nerves XI. A similar but weaker depressive effect was detected on the unitary EPSP resulting from stimulation of an ipsilateral cercal mechanoreceptor.This inhibition is attributed to multisynaptic inhibitory pathways impinging upon presynaptic excitatory neurones. The involvement of chloride ions is suggested by the observation that both picrotoxin and chloride-deficient salines abolished the inhibitory phenomenon. Presynaptic mannitolgap recording from cercal nerve XI revealed a chloride-dependent hyperpolarization in response to repetitive conditioning stimulation. The time course of this response was similar to that of presynaptic inhibition. Bath-application of GABA (20 mM) produced a chloride-dependent hyperpolarization followed by a depolarization of the intraganglionic part of the cerca-afferents. GABA-induced hyperpolarization and electrically-induced presynaptic hyperpolarization were both reversed in low chloride saline (166 mM chloride). It is proposed that presynaptic modulation of acetyl-choline release occurs at the cercal-afferent giant-interneurone synapses. The role played by GABA is duscussed.     

Induction of egg diapause in Bombyx mori by some cephalo-thoracic organs of the cockroach, Periplaneta americana

Induction of egg diapause in the silkworm, Bombyx mori by some cephalo-thoracic organs of the cockroach, Periplaneta americana was examined. All tissues tested such as brain, corpora cardiaca, corpora allata, suboesophageal and thoracic ganglia and nerve cords between thoracic ganglia were able to produce diapause eggs in non-diapause egg producers both by transplantation and injection of their crude homogenates. The homogenate of thoracic ganglia was effective even in pharate adults with the suboesophageal ganglion removed or in isolated abdomens of pharate adults.From these results, it was surmised that some endocrine organs, as well as the central nervous system in the cephalo-thorax of Periplaneta americana, contained the active principle responsible for egg diapause in Bombyx mori.     

Effect of opioid compounds on feeding and activity of the cockroach, Periplaneta americana

Opioid peptides have been implicated in regulation of feeding in invertebrates. Studies have suggested that receptors for opioids are present in cockroaches and that these receptors play roles in affecting both behaviour and feeding. We examined the effect of µ, δ, and κ opioid receptor agonists and antagonists on feeding, mass changes and activity in the cockroach, Periplaneta americana. The κ antagonist, nor-binaltorphimine, significantly increased food intake, while naltrexone (general antagonist) and naloxonazine (µ antagonist) both reduced feeding. A large mass loss was observed in cockroaches treated with nor-binaltorphimine, despite the increased food intake. Males did not lose as much mass during the 3 h as females, although drug treatment did have some effect on the loss. Time of activity (%) was not influenced by any drug. Water loss experiments suggested that nor-binaltorphimine increased water loss, accounting for the mass loss despite the increased feeding. We suggest that two populations of opioid receptors are present as previously reported, with one affecting feeding and the other involved with evaporative water loss.     

Ionophoretic application of acetylcholine on to the dendrites of an identified giant interneurone (GI 1) in the cockroach Periplaneta americana

Acetylcholine was ionophoretically applied within the neuropile of the sixth abdominal ganglion of the cockroach Periplaneta americana and depolarizing responses were recorded from an identified giant interneurone (GI 1) using the oil-gap, single-fibre technique. Estimated peak concentrations in the range 1.0 × 10^?11M ? 1.0 × 10^?8 M acetylcholine were present at the receptors, when this putative neurotransmitter was ionophoretically-applied, at threshold doses, to regions of the neuropile containing dendritic branches of GI 1. Using a stepping micro-drive unit for positioning the ionophoretic micropipette, it was shown that profiles of acetylcholine sensitivity correlated with the distribution of dendritic branches of GI 1 revealed by cobalt staining of the cell under test. It is concluded that acetylcholine receptors are widely distributed throughout the dendritic branches of GI 1 in the sixth abdominal ganglion. The results demonstrate the possibility of investigating the effects of locally-applied putative transmitter molecules and pharmacological agents on synaptic membranes of an identified neurone in the central nervous system of an insect.     

Neurophysiological changes associated with paralysis arising from body stress in the cockroach, Periplaneta americana

Two hours after physical stress, Periplaneta americana could be separated into three behavioural categories: normal to hyperactive; torpid with ataxia; and paralysed. At 2 hr, 68 per cent were either torpid or paralysed, at 20 hr, 83 per cent were paralysed. Weight loss was a distinct physiological symptom of stress paralysis: The calculated mean loss was 9·5 mg/hr for torpid insects and 12·4 mg/hr for paralysed cockroaches, losses were four to six times larger than those occurring in starved cockroaches. However, the haemolymph osrnolarities of the three categories showed no appreciable differences. Only starved and paralysed cockroaches showed a noticeable reduction in muscle fibre membrane potentials of the flexor tibia—a mean value below 40 mV for starved insects and a mean value below 50 mV for paralysed insects. Both of these categories consistently showed a lower amplitude for junctional potentials, but paralysed cockroaches showed a much higher incidence of complete failure to neural stimulation. Most muscle fibres of completely paralysed insects lost their sensitivity to direct extracellular stimulation while the loss in sensitivity was less evident in starved cockroaches. Axonal conduction on the crural nerve was not changed by stress, and the spontaneous efferent activity of completely paralysed insects was similar to the pattern of activity for normal cockroaches. Stress seemed to alter the volume and content of the intermyofibral spaces of muscles.     

The glycosaminoglycans of the thoracic ganglia of the nymphal stages of the cockroach, Periplaneta americana, and the locust, Locusta migratoria: A histochemical study

The glycosaminoglycans of the connective tissue matrices of the developing meso- and metathoracic ganglia of locusts and cockroach nymphs have been characterized. The neural lamella contains only chondroitin sulphate in the early nymphs, but gradually keratan sulphate accumulates in the later nymphs. The glial lacunar system cannot be detected histochemically in first instar locust nymphs, but it can be seen in the youngest cockroach nymphs; it is clearly visible in the older nymphs of both species. It contains only hyaluronate.A stereological analysis of the developing meso- and meta-thoracic ganglia of the cockroach shows that the relative volumes occupied by the neurones, neuropile, glial cells and glial lacunar system change during post-embryonic development.The physiological functions of the glycosaminoglycans in the neural lamella and glial system are discussed.