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

The stuctural 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 sort visual fibres. Horizontal fibre elements with branching patterns at different levels of the lamina apparently from three horizontal plexuses with presynaptic and/or postsynaptic connections to first-and second-order 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.     

Stress-induced release of octopamine in the American cockroach Periplaneta americana L

The level of arachidonic acid (AA) metabolites in the supernatants of cultured peritoneal exudate cells (PEC) were studied under various conditions using BCG and Corynebacterium parvum as stimulators. The metabolite levels were analyzed by thin layer chromatography (TLC). The degree of macrophage cytotoxic/cytostatic activity was dependent on the dose and character of stimulators used and the source of macrophages. The application of microcytotoxicity assay for the evaluation of tumor cell lysis (lung sarcoma SaL-1) in vitro revealed that peritoneal macrophages from healthy and tumor bearing BALB/c mice may affect the degree of antitumor response. In the supernatants of cultured PEC from tumor bearing mice AA level increased (by 10-fold) in comparison with PEC from healthy mice. Stimulation with BCG induced over a double level of AA in PEC isolated from tumor bearing mice nonstimulated or stimulated with C. parvum. A lower level of prostaglandins (PGs) was found in the supernatants of cultured PEC isolated from healthy mice (stimulated and non-stimulated), but the highest level of PGs was observed in the supernatants of cultured PEC isolated from tumor bearing mice stimulated with BCG. The unique metabolite of AA was found only in the supernatants from nonstimulated PEC from tumor bearing mice. PEC from tumor bearing mice produced metabolites of AA which were not detected in control group. These results suggest that macrophages also play a regulatory role by secretion of AA. This process can be modified by bacterial antigens.     

Locations and central projections of neurons associated with the retrocerebral neuroendocrine complex of the cockroach Periplaneta americana (L.)

Retrograde diffusion and precipitation of Co2+ reveals in the ipsilateral pars lateralis (PL) and contralateral pars intercerebralis (PI) of the brain neurons that enter the corpus cardiacum (CC), and, possibly, the corpus allatum (CA) on each side. The PL group consists of 29.6 +/- 8.4 somata that fill. Of these, 5.6 +/- 0.6 exceed 25 micrometer in diameter, 14.3 +/- 2.7 range from 15--25 micrometer, and 9.6 +/- 7.6 are smaller than 15 micrometer. After CoCl2 was applied to the right CC-CA of two males, 239 and 265 somata in the left PI stained. Except for 16 ranging from 30--45 micrometers and chiefly located anteriorly, a majority of these somata measured 10--25 micrometer. The only somata revealed by staining whole brains with the performic acid-resorcin fuchsin method are neurosecretory cells 10--20 micrometer in diameter located within the PI. In starved adult males there are 92.4 +/- 8.1 on the right, and 93.2 +/- 6.9 on the left. The largest somata in the PL group contain numerous granules that stain with paraldehyde fuchsin. These somata also fill with Co2+, and belong to neurosecretory cells that extend into the CC-CA. The cerebral distribution of branches from the PL group, and the relationship of these to the corpora pedunculata, central body, and arborizations from the PI decussation are described.     

Isolation of a novel RFamide peptide from the midgut of the American cockroach, Periplaneta americana

In insects (FM)RFamide-immunoreactive endocrine cells are ubiquitously present in the midgut, but the identity of the peptide(s) produced by these cells is unknown. The major RFamide-immunoreactive peptide from the midgut of the American cockroach, Periplaneta americana, was isolated and identified as Ala-Asn-Arg-Ser-Leu-Arg-Leu-Arg-Pheamide. This is a novel member of an arthropod peptide family, previously known only from mosquitoes and horseshoe crabs. Its abundance in the midgut suggests that it plays an important function in digestion.     

Fatty acids stimulate trehalose synthesis in trophocytes of the cockroach (Periplaneta americana) fat body

Trophocytes from the disaggregated fat body of the cockroach (Periplaneta americana) respond to synthetic hypertrehalosemic hormone (HTH) by increasing the rate of trehalose synthesis. The cells give a similar response when incubated with stearic, oleic, linoleic, or arachidonic acid. A maximal increase in trehalose synthesis was obtained with 1-10 microM fatty acids. Synthesis of trehalose by the trophocytes was also increased by 1 microM prostaglandin F2alpha to nearly the same extent as that evoked by HTH. Furthermore, the data show that the trophocytes are capable of converting linoleic acid into arachidonic acid. This suggests that the cells may convert arachidonic acid, formed from the linoleic acid released by the action of HTH, to a prostaglandin which serves as an integral part of the hypertrehalosemic mechanism.     

Learning in the intact cockroach (Periplaneta americana) when placed in a punishment situation.

Examined whether 40 intact cockroaches, Periplaneta americana, in a yoked-control punishment paradigm exhibit escape or avoidance learning, or both. Evidence for avoidance learning was unequivocal. Escape learning was not found. Asymptotic avoidance learning was found by 1.5 min. of training. The advantages of defining learning by experimental-control differences developing during training are discussed. (15 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Incubation effect in the cockroach (Periplaneta Americana).

Recorded in Exp I the activity of 72 cockroaches in an open field in which they had received footshock (FS) 1, 3, 10, 30, or 120 min before. The FS produced an incubation effect, i.e., a general decrease in activity, with the greatest decrease occurring at the longer intervals. In Exp II, activity of 77 cockroaches was recorded in an open field 120 min after 1 FS and 1, 3, 10, or 30 min after a 2nd. When no 2nd FS was administered, the cockroaches were inactive; however, when testing occurred within 10 min of the 2nd FS, activity was greatly increased. This gradient of hyperactivity paralleled exactly the incubation effect demonstrated in Exp I. Results of both experiments are almost identical to the results of comparable experiments in which rats have served as Ss. Thus, the incubation effect is an adaptive behavior pattern resulting from the transient activating effects of noxious stimulation. (29 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Characterization of neurons of the nodose ganglion having constant impulse activity

The present studies were undertaken to characterize selenium distribution in egg white. Ion-exchange chromatography fast protein liquid chromatography (FPLC) and flow injection atomic (absorption) spectrometry (FIAS) were used to separate egg white proteins and to determine the selenium content of different fractions. After purification, nine different proteins were identified with sodium dodecyl sulphate-polyacrylamide gel electrophoresis and 56% of the total selenium content was found to be associated with ovalbumin-1 and -2 (+/- 500 ng/g), which is the main protein in egg white. Flavoprotein was determined to be the richest selenium-containing protein (1800 ng/g). The selenium content of the other proteins (lysozyme, conalbumin, globulins and ovomucoid) ranged from 359 to 1094 ng/g.     

Influence of factors released from sciatic nerve on adult dorsal root ganglion neurons

Previous experiments have shown that medium conditioned (CM) by denervated peripheral nerve contains a process outgrowth promoting factor(s) for cultured adult frog dorsal root ganglion (DRG) neurons. The present experiments further characterize the influences of these factors on DRG neurons. The growth factors increases average process length by threefold, restricts the number of processes extended from four to two while simultaneously altering the morphology of those processes. Neurons with preexisting processes respond to the factors by significantly increasing the length of 35% of these processes. Only the newly elongated portions of preexisting processes have a morphology typical of factor-induced processes, while the previously extended portions retain their original morphology. The number of processes of these neurons remains unchanged. Although composed of two populations according to size, neurons in both populations are similarly influenced, suggesting that the factors influence neurons of all sensory modalities. To look at other possible influences of the nerve-released factors, a novel simple culture system has been developed in which concentration gradients of these factors can be established and maintained. The front of the outgrowth-promoting influence in these cultures could be followed over time (up to 9 days) as it affected the process length and morphology of neurons at increasing distances (up to 8 mm) from the source of the factors. The trophic factors may play important roles during regeneration in vivo by influencing the cytoskeletal organization in the cell body and growth cones to bring about a stabilization and consolidation of growth cone membrane of only a limited number of processes resulting in increasing the rate of process elongation. The factors may also serve to direct process outgrowth, which can be examined using the new culture system.     

Mushroom bodies of the cockroach: activity and identities of neurons recorded in freely moving animals

This article describes novel attributes of the mushroom bodies of cockroaches revealed by recording from neurons in freely moving insects. The results suggest several hitherto unrecognized functions of the mushroom bodies: extrinsic neurons that discriminate between imposed and self-generated sensory stimulation, extrinsic neurons that monitor motor actions, and a third class of extrinsic neurons that predict episodes of locomotion and modulate their activity depending on the turning direction. Electrophysiological units have been correlated with neurons that were partially stained by uptake of copper ions and silver intensification. Neurons so revealed correspond to Golgi-impregnated or Lucifer yellow-filled neurons and demonstrate that their processes generally ascend to other areas of the protocerebrum. The present results support the idea of multiple roles for the mushroom bodies. These include sensory discrimination, the integration of sensory perception with motor actions, and, as described in the companion article, a role in place memory.     

Lectin histochemistry of the metathoracic ganglion of the locust Schistocerca gregaria before and after axotomy of the tympanal nerve

The vascular endothelium is involved in the production of many important substances which are involved in a cardiovascular pathophysiology. One such substance which is synthesised by, and stored in, endothelial cells is von Willebrand factor (vWf). When released, vWf appears to mediate platelet aggregation and adhesion. Numerous clinical and experimental reports suggest that high vWf levels reflect damage to the endothelium or endothelial dysfunction. The close association between vWf and the processes of thrombus formation (thrombogenesis) or atherogenesis also suggests that high vWf levels may be a useful indirect indicator of atherosclerosis and/or thrombosis. The availability of a useful marker of endothelial dysfunction may have potential clinical value. The measurement of such a marker can perhaps be a non-invasive way of assisting in clinical diagnosis or as an indicator of disease progression. High vWf levels have also been shown to have prognostic value in patients with ischaemic heart disease, peripheral vascular disease and inflammatory vascular disease. However, there is limited information that increased vWf is actually casual in the progression of vascular disease and that measures aimed at reducing vWf levels will be beneficial. In addition, interpretation of raised plasma vWf levels is complicated by the fact that vWf may be an acute phase reactant. Further research is indicated to explore the predictive value of this marker in population studies and, perhaps, therapeutic approaches in (and the value of) modifying vWf levels or function.     

Effect of diflubenzuron on the incorporation of UDP-N-acetyl-[3H]glucosamine (UDP-[3H]NAGA) to chitin in permeabilized, and isolated integuments from the newly molted American cockroach Periplaneta americana

1. Diflubenzuron (DFB) was found to inhibit the incorporation of UDP-N-acetyl-[3H]glucosamine (UDP-[3H]NAGA) to chitin in permeabilized and isolated integument from newly molted American cockroach. 2. The favorable experimental conditions demonstrating the effect of diflubenzuron were: 10 mM phosphate, low calcium concentration (10(-6) M-10(-8) M), high potassium concentration (> 100 mM), and high pH (> or = 7). 3. The action of diflubenzuron was completely erased by preincubating the isolated integument with valinomycin, FCCP, or A23187. 4. By lowering the external pH to 5.2, it was also possible to reduce the rate of UDP-[3H]NAGA incorporation to the extent that DFB's effect was no longer recognizable. 5. Both Cs+ and Rb+ could replace K+ in maintaining a high level of chitin synthesis and the inhibitory action of DFB under the optimum conditions.     

Effects of intraventricular and epicardial application of adenosine on electrical activity of medullary PGL neurons

The effects of intraventricular injection and epicardial application of adenosine on spontaneous electrical activity of nucleus paragigantocellularis lateralis (PGL) neurons in rostral ventrolateral medulla (RVLM) were examined in 35 anesthetized rats with sinoaortic denervation and vagotomy. The results obtained were as follows: (1) The spontaneous discharge of 121 PGL neurons (mean discharge rate: 22.5 +/- 1.9 spikes/s) were recorded in 35 rats. (2) In response to intraventricular injection of adenosine (0.5 mumol/kg), mean arterial pressure (MAP) was initially increased by 1.7 +/- 0.2 kPa(P < 0.001) and subsequently decreased by 4.6 +/- 0.5 kPa(P < 0.001), while the heart rate (HR) was decreased by 126.5 +/- 12.3 bpm (P < 0.001). Of 35 PGL spontaneous discharge units responsive to intraventricular injection of adenosine, 30 showed an average increase from 21.9 +/- 2.6 to 29.2 +/- 3.4 spikes/s (P < 0.001), 3 with no change, while 2 with a decrease. (3) Following epicardial application of 20 mmol/L adenosine, the BP and HR were not significantly changed, while the spontaneous discharge of 22 PGL neurons were increased from 18.8 +/- 1.9 to 26.9 +/- 2.8 spikes/s (P < 0.001), and that of 3 neurons was not changed. (4) The excitatory response of PGL neurons to intraventricular injection or epicardial application of adenosine was completely inhibited by pretreatment with selective adenosine A1-receptor antagonist 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX, 500 micrograms/kg). (5) Following epicardial application of phenol or bilateral stellate ganglionectomy, adenosine failed to affect the activity of PGL neurons. The results obtained indicate that adenosine may stimulate cardiac sympathetic afferents through adenosine A1-receptor, thereby resulting in the activation of PGL neurons.     

Influence of endogenous GM1 ganglioside on TrkB activity, in cultured neurons

To establish direct linkage between the ethanol-inducible cytochrome P450, CYP2E1, ethanol hepatotoxicity, and lipid peroxidation, a HepG2 cell line which expresses human CYP2E1 was established by retroviral infection. Ethanol produced a time-and concentration-dependent cytotoxicity to HepG2 cells expressing the CYP2E1 but not to control cells. The ethanol toxicity was prevented by inhibitors of CYP2E1 and antioxidants. In a similar manner, addition of a polyunsaturated fatty acid such as arachidonic acid produced toxicity to the cells expressing CYP2E1 but not the control cells. Toxicity was associated with enhanced lipid peroxidation and was prevented by antioxidants. The ethanol and arachidonic acid toxicity was apoptotic in nature and was associated with activation of Caspases I and III. The toxicity and apoptosis could be prevented by peptide inhibitors of ICE and by transfection with a plasmid containing the cDNA for human Bcl-2. These results show that this HepG2 cell model can be used to establish a CYP2E1-dependent ethanol hepatotoxicity system, and that induction of a state oxidative stress appears to play a central role in the CYP2E1-dependent apoptosis and cytotoxicity.     

Vestibular ganglion neurons survive the loss of their cerebellar targets

Neuronal survival during mammalian development crucially depends on target-derived neurotrophic factors. Target loss removes this trophic support and leads in most cases to the transsynaptic retrograde degeneration of the respective afferents. Primary vestibular afferents (PVA) originating from bipolar neurons in the vestibular ganglion (VG) are the first mossy fibers that enter the cerebellum, but little is known about the survival requirements of VG neurons. In the present study the influence of the differential granule cell (GC) target loss on the survival of VG neurons was studied quantitatively using unbiased stereological methods in the cerebellar mutants Purkinje cell degeneration (pcd/pcd), Lurcher (Lc/+), and Weaver (wv/wv). Neither the secondary GC loss in the Purkinje cell deficient mutants pcd/pcd and Lc/+, nor the primary loss of GCs in wv/wv produced any significant reduction in the total number of bipolar neurons in the VG compared to controls. So, PVA neurons are highly resistant to cerebellar target deprivation and survive in the absence of cerebellar granule and Purkinje cells, regardless of whether the target loss occurs before (in wv/wv), during (in Lc/+) or after (in pcd/pcd) the mossy fiber-granule cell synaptogenesis.     

Quantitative structure-activity studies of octopaminergic 2-(arylimino)thiazolidines and oxazolidines against the nervous system of Periplaneta americana L

The quantitative structure-activity relationship (QSAR) of octopaminergic 2-(arylimino)thiazolidines (AITs) and 2-(arylimino)oxazolidines (AIOs) against the thoracic nerve cord of the American cockroach, Periplaneta americana L., was analysed using reported physicochemical parameters and regression analysis. The more electron-donating, the less bulky at m-position, and the more hydrophobic the substituent, the greater the activity. The plots of observed log Vmax values against calculated log Vmax values having substituents on the m-position deviated downwards from those of compounds having substituents at the 0- and/or p-positions. The more hydrophobic and the more electron-withdrawing the substituent, the greater the activity. AIO with a 2, 3, 4-trichlorophenyl group (58) was more active than its thiazolidine derivative, 2-(2,3,4-trichlorophenylimino)thiazolidine (38) in terms of Vmax:Vmax of 58 was 30% relative to octopamine (OA), whereas that of 38 has been 9% relative to OA, respectively. Superimposition of energy-minimized OA and 58 revealed structural and conformational similarities that might account for the high activity of 58.     

Regeneration of cercal filiform hair sensory neurons in the first-instar cockroach restores escape behavior

Neural regeneration in the escape circuit of the first-instar cockroach is described using behavioral analysis, electrophysiology, intracellular staining, and electron microscopy. Each of the two filiform hairs on each of the animal's cerci is innervated by a single sensory neuron, which specifically synapses with a set of giant interneurons (GIs) in the terminal ganglion. These trigger a directed escape run. Severing the sensory axons causes them to degenerate and perturbs escape behavior, which is restored to near normal after 4-6 days. Within this time, afferents regenerate and reestablish arborizations in the terminal ganglion. In most cases, regenerating afferents enter the cercal glomerulus and re-form most of the specific monosynaptic connections they acquired during embryogenesis, although their morphology deviates markedly from normal; these animals reestablish near normal escape behavior. In a few cases, regenerating afferents remain within the cercus or bypass the cercal glomerulus, and thereby fail to re-form synapses with GIs; these animals continue to exhibit perturbed escape behavior. We conclude that in most cases, specific synapses are reestablished and appropriate escape behavior is restored. This regeneration system therefore provides a tractable model for the establishment of synaptic specificity in a simple neuronal circuit.     

Giant input neurons of the mushroom body: intracellular recording and staining in the cockroach

Covalent complexes of nucleic acids and proteins are widespread among viruses. Covalent complexes of RNA and proteins are proposed to exist in eukaryotic cells. The goal of this work was to obtain specific antibodies to the covalent linkage unit (CLU) between virus RNA and protein to search cellular RNA-protein complexes. Antibodies were generated by direct immunization of a rabbit with the BSA-coupled EMC virus RNA-VPg complex. By a dot-blot immunoassay and immunofluorescent microscopy it was found that the antibodies specifically recognize both EMC virus RNA-VPg and synthetic CLU-containing compounds. Thus, a fraction of the antibodies was directed to CLU.