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Inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase lower fecundity in the German cockroach: correlation between the effects on fecundity in vivo with the inhibition of enzymatic activity in embryo cells.

The enzyme 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase is crucial to insect development and reproduction, as revealed by the sterilising properties of some specific inhibitors of it. In the present paper, we study the sterilising effects of a number of HMG-CoA reductase inhibitors on the German cockroach, Blattella germanica (L). The inhibitors tested were naringenin, lovastatin, mevastatin, simvastatin, atorvastatin and fluvastatin. The first two compounds were ineffective or scarcely effective as HMG-CoA reductase inhibitors. The most active compounds in vivo were fluvastatin and atorvastatin, followed by simvastatin and mevastatin. They were equally ranked when tested as HMG-CoA reductase inhibitors in the B germanica embryonic derived cell line UM-BGE-1. This suggests that this cell line may be an appropriate tool for testing HMG-CoA reductase inhibitors and so to predict their properties as insect sterilising agents with insecticide potential.

Animals↗

Transient appearance and localization of a 26-kDa lectin, a novel member of the Periplaneta lectin family, in regenerating cockroach leg.

A lectin consisting of a 26-kDa subunit was purified from the hemolymph of adult American cockroaches (Periplaneta americana) and shown to be a member of the Periplaneta lectin family. Immunoblotting analysis revealed that this lectin appeared transiently in regenerating legs after nymphal legs were amputated. Thus, its mode of appearance was similar to that of regenectin, another member of the Periplaneta lectin family. However, its time of appearance and localization in the regenerates were different from those of regenectin: the 26-kDa lectin appeared in regenerates in the middle stage of regeneration and was localized in the vicinity of the outside of newly formed epidermal cells. These results suggest that this lectin participates in the organization or stabilization of the epidermis during leg regeneration.

Animals↗

Comparison of defolliculated oocytes and intact follicles of the cockroach using the vibrating probe to record steady currents.

Follicle cells were removed by dissection from early vitellogenic oocytes of the cockroach Blattella germanica. The vibrating probe was used to record steady currents from 19 defolliculated oocytes and 19 intact follicles of the same developmental stage. Defolliculated oocytes generated currents that were stable and distinguishable (by intensity or selective direction) from background reference values. Distributions of the intensities of reference values and experimental values were, in general, similar in both intact and defolliculated preparations. The patterns of currents generated by preparations recorded in the mid-sagittal plane were analyzed for both defolliculated oocytes (n = 8) and intact follicles (n = 10). The larger, generally more mature preparations in both groups generated patterns of current similar to the pattern seen in mid-vitellogenic follicles (focused inward near the germinal vesicle (GV), the presumptive ventral side, and broadly outward on the apo-GV side, the presumptive dorsal side). Smaller sized preparations in both groups showed inward or outward current on the apo-GV aspect and, typically, inward current at the GV. Only two defolliculated oocytes, and no intact follicles, appeared to generate outward current at the GV, and we believe this observation resulted from recording slightly outside the mid-sagittal plane. We conclude that preparations during early-vitellogenesis initially generate currents without an asymmetric pattern and that the inward flux at the GV is the first step in developing patterns of currents. The results suggest that the oocyte (and not the follicle cell epithelium) is responsible for generating the various patterns of currents observed in early-vitellogenic stages. At the end of early-vitellogenesis, the follicle cell epithelium begins to adhere tightly to the oocyte. The possibility is considered that the follicle cells may influence the currents generated during mid-vitellogenesis.

Animals↗

In vitro hormonal regulation of glycogen phosphorylase activity in fat body of the tropical cockroach, Blaberus discoidalis.

Blaberus hypertrehalosemic hormone (Bld-HTH)-dependent glycogen phosphorylase activation was investigated using in vitro fat bodies from the cockroach, Blaberus discoidalis. Resting levels of active phosphorylase were decreased by the presence of trehalose and glucose. Phosphorylase activation was dose-responsive to Bld-HTH and increased ca. 3-fold over a range of 0.02 to 2 nM Bld-HTH. Maximum phosphorylase activation required only 5-min exposure to Bld-HTH; reversion to the inactive state began within 15 min after Bld-HTH removal and was completed by 60 min. Octopamine also activated phosphorylase but required 10(3)-fold higher concentrations than did Bld-HTH. Concentrations of Bld-HTH and octopamine that increased active phosphorylase did not elevate fat body cAMP levels, although a high concentration of octopamine increased tissue cAMP levels. cAMP did not increase phosphorylase activity, but Ca2+ was important for both Bld-HTH- and octopamine-dependent phosphorylase activation.

Animals↗

Calcium-dependent signal transduction by the hypertrehalosemic hormone in the cockroach fat body.

The hypertrehalosemic hormones (HrTH) are insect peptides that stimulate the fat body to synthesize trehalose. The present research investigated the relative importance of extra- and intracellular Ca2+ for HrTH signal transduction in the fat body of the cockroach, Blaberus discoidalis. Although trehalose synthesis could be stimulated significantly by B. discoidalis HrTH (Bld-HrTH) in the absence of Ca2+, the presence of Ca2+ was necessary to attain a maximum hormonal response. Entry of extracellular Ca2+ mediated by ionophores (A23187, ionomycin) produced no significant hypertrehalosemic effects, whereas release of intracellular Ca2+ by thimerosal or thapsigargin significantly stimulated trehalose synthesis. The hypertrehalosemic effect by thimerosal occurred independently of extracellular Ca2+. Dantrolene inhibits intracellular Ca2+ release, and its presence partially suppressed Bld-HrTH-dependent hypertrehalosemia. In conclusion, the presence of either extracellular Ca2+ or intracellular Ca2+ enhanced the Bld-HrTH response, but agents that affected intracellular Ca2+ release produced the more conclusive effects.

Animals↗

Cockroach allatostatin-like immunoreactivity in the central nervous system of the freshwater snails Bulinus globosus (Planorbidae) and Stagnicola elodes (Lymnaeidae).

Allatostatin-like immunoreactivity was demonstrated in the central nervous system (CNS) neurons of the freshwater pulmonate snails Bulinus globosus (Planorbidae) and Stagnicola elodes (Lymnaeidae). Immunopositive neurons were localized using a monoclonal antibody highly specific to allatostatin I (APSGAQRLYGFGL-amide) from the cockroach Diploptera punctata. All CNS ganglia in both snail species contained immunopositive neurons. The pedal ganglia showed large numbers (80-100) of neurons in Bulinus and Stagnicola. Large numbers of immunopositive cells were also found in the cerebral (60) and buccal ganglia (20) of Bulinus. Other ganglia contained fewer immunopositive cells, but these cells were most concentrated in the cerebral (Stagnicola) and left parietal (Bulinus) ganglia and the visceral ganglia of both species. The high concentration of immunopositive cells in the pedal ganglia and axons demonstrable in the pedal nerves suggests that one possible function for a molluscan allatostatin-like peptide would be to modulate muscular function. Extract of Stagnicola CNS effected 50% inhibition of juvenile hormone synthesis by corpora allata of D. punctata at between 15 and 30 CNS equivalents, providing further evidence that the molluscan immunoreactive material is a peptide, or peptides, with sequence similarity to the active part of the D. punctata allatostatins.

Animals↗

The effect of biogenic amines and their analogs on carbohydrate metabolism in the fat body of the cockroach Blaberus discoidalis.

Several biogenic amines and their analogs were examined for stimulatory effects on glycogen phosphorylase activity and trehalose biosynthesis in fat body of the cockroach, Blaberus discoidalis. Octopamine and synephrine were the most potent activators of fat body phosphorylase; 10 muM octopamine being nearly as effective as the hypertrehalosemic hormone (HTH). Epinephrine, norepinephrine, and tyramine produced intermediate effects, whereas dopamine, 5-hydroxytryptamine, and melatonin had no effect. The fat body octopamine receptors appeared to be pharmacologically related to vertebrate alpha-adrenergic receptors and belonged to the Octopamine1 class receptor. In contrast to previous reports, synephrine also induced both phosphorylase activation and hypertrehalosemia as effectively as octopamine. Demethylchlordimeform, a formamidine insecticide structurally similar to octopamine, also strongly activated fat body phosphorylase, possibly by interaction with the octopamine receptor.

Amidines↗

Light and electron microscopic study of a Rickettsiella species from the cockroach Blatta orientalis.

An infection with Rickettsiella sp. was responsible for an illness causing heavy body swelling in the Oriental cockroach Blatta orientalis. Reproduction of the colony stagnated. Vacuoles with parasitic bacteria occurred mainly in the fat body, but also in nearly all other organs, such as gut epithelium, Malpighian tubules, blood cells, and ovarioles. The parasites clearly differed from the symbiotic bacteria of the genus Blattabacterium, which regularly occur in the mycetocytes of B. orientalis. The vacuoles contained four stages of Rickettsiella: (1) infectious, electron-dense, rod-like elementary bodies (mean size 300 x 145 nm); (2) an electron-dense, flat intermedium stage, called flat body (mean size 515 x 255 x 125 nm); (3) an electron-light, spherical intermedium stage, called condensing sphere (mean size 340 nm); portions of cytoplasm condensed crescent-like at the border or in the center of the cell; and (4) large, spherical, electron-light initial bodies that multiplied by binary fission (mean size 600 nm). The initial bodies had a three-layered cell boundary, but all other stages had a five-layered cell boundary. Elementary and flat bodies contained an electron-light, oblique lamella and an oval structure with an array of ribosome-like granules, respectively. In contrast to other species of Rickettsiella, crystal formation or multiple division did not occur. The described species of Rickettsiella is different from "R. blattae," which belongs to the R. popilliae group. Instead, it shares more similarities with the R. chironomi group. To avoid confusion, it was provisionally named "R. crassificans."

Animals↗

The in vivo regulation of pioneer axon growth by FGF-2 and heparan sulfate proteoglycans in cultured embryos of the cockroach.

Antibody perturbation experiments on cultured cockroach embryos demonstrated that a localized source of an FGF-2-like immunoreactive molecule in the head is required for the proper growth of pioneer axons in the leg. The study of axon growth in various fragments of cultured embryos and in the presence of various conditioned media showed that FGF-2 is needed to counteract the effects of an inhibitor of axon growth produced in the body trunk of the embryo. Endogenous heparan sulfate proteoglycans mediate these effects of FGF-2 on axon growth. The results of experiments with FGF-2 and/or body trunk axon growth inhibitor added to the culture medium indicate that more globally and uniformly distributed molecules may play as important a role in axon guidance as the more spatially restricted guidance cues. The results are interpreted in terms of a model that is consistent with a role for the FGF-2 receptor in axon growth.

Animals↗

The effects of methylglyoxal on central synaptic transmission in the isolated nerve cord of the cockroach. (Periplaneta americana L).

Methylglyoxal (10(-5) to 1.5 X 10(-4) M) was found to have excitatory effects on synaptic transmission in the isolated 6th abdominal ganglion of the cockroach. There was a concentration-dependent depolarization of the giant interneurones which was accompanied by an increase in the amplitude and duration of electrically evoked excitatory postsynaptic potentials. The frequency of spontaneous activity was also increased.

Aldehydes↗

Synaptic specificity in the first instar cockroach: patterns of monosynaptic input from filiform hair afferents to giant interneurons.

Direct evidence for monosynaptic connections between filiform hair sensory axons and giant interneurons (GIs) in the first instar cockroach, Periplaneta americana, was obtained using intracellular recording and HRP injection followed by electron microscopy. GIs 1-6 all receive monosynaptic input from at least one filiform afferent axon. GI1, GI2 and GI5 receive input only from the medial (M) axon, while GI3, GI4 and GI6 receive input from both M and lateral (L) axons. The dendrites of GI3 and GI6 which are contralateral to the cell bodies receive input from both axons whereas the smaller ipsilateral dendritic fields have synapses only from the L axon. GI5 has M axon input only onto its contralateral dendrites. In 50% of preparations GI7 receives weak input from the ipsilateral L axon. There is no obvious relationship between the morphology of the giant interneurons and the pattern of input they receive from the filiform afferents.

Animals↗

Filter characteristics of cercal afferents in the cockroach.

The response dynamics of cercal afferents in the cockroach. Periplaneta americana, were determined by means of a cross-correlation technique using a Gaussian white noise modulation of wind as a stimulus. The white noise stimulus could evoke sustained firing activity in most of the afferents examined (Fig. 1). The spike discharges were unitized and then cross-correlated with the stimulus to compute 1st- and 2nd-order Weiner kernels. The 1st-order kernels from a total of 28 afferents were biphasic and closely matched the time differential of a pulse (Figs. 1, 3 and 4). The amplitude and waveform of the kernels depended on the stimulus angle in such a way that the kernels were the mirror image of those on the polar opposite side (Figs. 2 and 3). The 2nd-order kernels were also differential. They had 2 diagonal peaks and 2 off-diagonal valleys in a 2-dimensional plot with 2 time axes (Figs. 1, 5 and 6). This 4-eye configuration was basically invariant irrespective of the stimulus angle, although the kernels varied in amplitude when the stimulus angle was changed. The time between the peak and a following trough of the 1st-order kernel was constant and had a mean of 4.6 +/- 0.1 ms, whereas the time between 2 diagonal peaks of the 2nd-order kernels was 4.7 +/- 0.1 ms (Figs. 4 and 6), suggesting that wind receptors (filiform sensilla) on cerci act as a band-pass filter with a peak frequency of about 106 Hz. The peak time, however, varies from 2.3 to 6.9 ms in both kernels, which may reflect the spatial distribution of the corresponding hairs on the cercus. The summation of the 1st- (linear) and 2nd-order (nonlinear) models precisely predicted the timing of the spike firing (Fig. 8). Thus, these 2 lower-order kernels can totally characterize the response dynamics of the wind receptors. The nonlinear response explains the directional sensitivity of the sensory neurons, while the differentiating 1st-order kernel explains the velocity sensitivity of the neurons. The nonlinearity is a signal compression in which one of the diagonal peaks of the 2nd-order kernel always offsets the downward phase of the 1st-order kernel (Fig. 7) and obviously represents a half-wave rectification property of the wind receptors that are excited by hair movement in only one direction and inhibited by hair movement in the polar opposite direction.

Animals↗

Circadian changes in cockroach ommatidial structure.

1. Morphological correlates of circadian changes in eye sensitivity to light measured electrophysiologically were sought in the cockroach, Leucophaea maderae. Cross sections of ommatidia removed at subjective midday and subjective midnight on 3 successive days from roaches held under constant darkness (DD) at 25 +/- 2 degrees C were examined using a transmission electron microscope for morphological differences related to sampling time. 2. The temporal difference in submicrovillar cisternae (SMC) area appeared to exhibit a circadian rhythm, however, the amplitude of this temporal difference measured under DD was less than that observed under LD 12:12 conditions. SMC areas characteristic at nighttime were achieved at subjective midnights but the area diminished only partially toward the daytime state on subjective middays. 3. Rhabdom area remained constant and the daily rhythm of screening pigment granules (SPG) arrangement about the rhabdom was not observed under conditions of constant darkness. 4. Results of this study indicate that a pacemaker(s) actively influences the change in the SMC toward the nighttime state, whereas, the change toward the daytime state results from a passive mechanism that possibly could be accelerated by light.

Animals↗

Intracellular nonlinear frequency response measurements in the cockroach tactile spine neuron.

The threshold of the cockroach tactile neuron increases strongly with depolarization by a process involving at least two time constants. This effect is probably responsible for the rapid and complete adaptation of the neuron's response to step inputs. A technique for intracellular recording and stimulation of the neuron has recently been established and this allows direct observation of the dynamic response of the neuronal encoder. A white noise stimulus was used to modulate the membrane potential of the neuron. The first-order frequency response function between membrane potential and action potential discharge could be explained by a variable threshold model with two time constants. Second-order frequency response functions could be accounted for by a Wiener cascade model. The dynamic nonlinear behavior of the encoder can therefore be explained by a unidirectional threshold which increases linearly and dynamically with membrane potential.

Acoustic Stimulation↗

Nonlinear neuronal mode analysis of action potential encoding in the cockroach tactile spine neuron.

Neuronal mode analysis is a recently developed technique for modelling the behavior of nonlinear systems whose outputs consist of action potentials. The system is modelled as a set of parallel linear filters, or modes, which feed into a multi-input threshold. The characteristics of the principal modes and the multi-input threshold device can be derived from Laguerre function expansions of the computed first- and second-order Volterra kernels when the system is stimulated with a randomly varying input. Neuronal mode analysis was used to model the encoder properties of the cockroach tactile spine neuron, a nonlinear, rapidly adapting, sensory neuron with reliable behavior. The analysis found two principal modes, one rapid and excitatory, the other slower and inhibitory. The two modes have analogies to two of the pathways in a block-structured model of the encoder that was developed from previous physiological investigations of the neuron. These results support the block-structured model and offer a new approach to identifying the components responsible for the nonlinear dynamic properties of this neuronal encoder.

Action Potentials↗

A nonlinear model of step responses in the cockroach tactile spine neuron.

Rapid sensory adaptation in the cockroach tactile spine neuron has previously been associated with a labile threshold for action potentials, which changes with the membrane potential by a process involving two time constants. A feed-forward, variable-threshold model has previously been used to account for the frequency response function of the neuron when stimulated with small-signal, white-noise currents. Here, we used a range of accurately controlled steps of extracellular current to stimulate the neuron. The same model was able to predict the individual step responses and could also fit the entire set of step responses from a single neuron if an initial, saturating, static nonlinearity was included. These results indicate that the two-time-constant, variable-threshold model can account for most of the rapidly adapting behavior of the tactile spine neuron.

Animals↗

Walking in the American cockroach: the timing of motor activity in the legs during straight walking.

The timing of bursts of motor activity in extensor muscles in the coxae of pairs of legs in intact freely walking American cockroaches was studied. The timing of bursts in adjacent and non-adjacent leg pairs generally reflected the common alternating tripod gait of these insects. Detailed study of the timing further revealed two previously unreported features. 1) The timing of extensor bursts in the middle legs relative to bursts in the rear legs was more variable than it was relative to those in the front legs. This difference in variability was statistically significant for the means of bursts when all insects were considered together as well as for bursts in individual insects. An apparent difference in variability of the timing of burst starts compared to burst ends for any one leg pair was not significant. 2) There was a shift in the timing of motor bursts relative to one another when an insect walked fast such that motor bursts in the middle legs tended to lag farther behind those in the front legs, and those in the rear legs tended to lag farther behind those in the middle legs compared to the timing during slow walking. This shift was apparent in both burst starts and burst ends, although more obvious in the former. It occurred in both ipsilateral and contralateral leg pairs, and in both the mean data and the data for individual insects. The implications of these characteristics of the timing data are discussed in terms of the neural organization of insect walking.

Animals↗

Localization of pancreatic polypeptide (PP)-like immunoreactivity in the central and visceral nervous systems of the cockroach Periplaneta.

The central and visceral nervous systems of the cockroach Periplaneta americana were studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibody to bovine pancreatic polypeptide (PP). PP-like immunoreactive neuron somata are most numerous in the brain; at least 6 pairs of cell groups occur in clearly defined regions. Three pairs of cells each are also present in the suboesophageal ganglion and the thoracic ganglia, one pair of single cell each in the first abdominal and the frontal ganglia, and 4 to 6 pairs of single cells in the terminal ganglion. No reactive cells were found in the retrocerebral complex and the second to the fifth abdominal ganglia. The axons containing PP-like immunoreactivity issue many branches that are distributed in the entire brain-retrocerebral complex, ventral cord, and visceral nervous system. PP-like immunoreactive material produced in the brain seems to be transported by three routes: protocerebrum to corpora cardiaca (-allata) through the nervi corporis cardiaci, tritocerebrum to visceral nervous system through frontal commissures, and to ventral cord through circumoesophageal connectives. A possible homology between the mammalian brain-GEP (gastro-entero-pancreatic) system and the brain-midgut system of this insect is discussed.

Animals↗