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PubMed1981

Morphological changes in gland cells and axons resulting from stimulation of the salivary nerves of the cockroach, Nauphoeta cinerea.

The salivary glands of the cockroach, Nauphoeta cinerea (Oliver, 1789), are innervated and there is considerable evidence to suggest that dopamine is the neurotransmitter at the neuroglandular junction. As the gland is a bilaterally symmetrical structure it was possible to electrically stimulate the salivary nerve supplying the ipsilateral side of the gland whilst the contralateral side of the gland served as a convenient control. Saliva elicited from the glands by electrical stimulation of these nerves was collected and used to monitor the physiological state of the tissue. Glands were fixed for light and electron microscopy during secretion and it was observed that the ductules in peripheral acinar cells were distended in stimulated sides of the glands but not in contralateral unstimulated sides. This evidence implies that peripheral cells are responsible for the initiation of salivary fluid secretion. Changes were also observed in the catecholamine containing axons that innervate the glands. In stimulated axons a statistically significant reduction in numbers of small agranular vesicles was observed when compared with contralateral unstimulated controls and freshly fixed tissue. This was not the case with the larger granular vesicles of the same axons which showed no reduction in number as a result of stimulation. In addition it was also noted that the small agranular vesicles tended to aggregate and change their shapes in response to nerve stimulation. These results imply that the small agranular vesicles play a role in transmitter release.

Animals↗
PubMed1982

Morphological differences between excitatory and inhibitory nerve terminals in cockroach coxal muscles.

Two morphologically distinct types of neuromuscular junction on the coxal leg muscles of the cockroach, Periplaneta americana, which have been physiologically described as innervated by fast, slow and inhibitory nerve fibers, have been found. In one type of neuromuscular junction the axon terminal contains many round clear synaptic vesicles and contacts several sarcoplasmic extensions from the muscle fiber. The muscle processes adhere to the axon terminal for a short distance (short contact or SC type). The axon terminal of the other type of neuromuscular junction directly contacts the muscle fiber and no extensions of the muscle fiber are formed. The contact region is comparatively long (long contact or LC type). The nerve terminal contains many polymorphic synaptic vesicles. From a correlation of the present morphological findings and the previous physiological results, it may be suggested that the SC type of nerve terminal represents both fast and slow nerve terminals and the inhibitory terminal is of the LC type.

Animals↗
PubMed1989

Immobilizing and lethal effects of spider venoms on the cockroach and the common mealbeetle.

Immobilizing and lethal effects of the venoms obtained from six spider species (Brachypelma albopilosum, Atrax robustus, Cupiennius salei, Selenops mexicanus, Tegenaria atrica, Argiope bruennichi) were tested on Blatta orientalis (cockroach) and Tenebrio molitor (common mealbeetle). The immobilizing effects were quantified by measuring insect locomotor activity in circle arenas observed over 72 hr after venom injection. Both insect species showed cramps, quivering and jerking of the limbs as well as flaccid paralysis after venom injection. Through relative toxicity of the venoms tested is the same in T. molitor and B. orientalis, T. molitor is absolutely less sensitive to spider venoms. The effects on locomotor activity show time characteristics specific for each venom. A dependence of the venom paralyzing effects on insect locomotor activity, low intensity of the initial excitatory phase of the venom effects and partial recovery of the insects was found with A. bruennichi and T. atrica venom. The maximal venom yields of A. bruennichi and S. mexicanus are not lethal to B. orientalis, indicating that the mere immobilizing effects of spider venoms are far more crucial to prey capture than their lethal effects. The contribution of a variety of differently acting neurotoxic components in spider venoms to the observed venom effects on insects and the significance of the venoms in spider nutrition, hunting behaviour and ecology are discussed.

Animals↗
PubMed1981

Effects of cycloheximide and puromycin on learning and retention in the cockroach, P. americana.

A new one-session T-maze training procedure for cockroaches, in which animals were trained to turn right or left to avoid shock, is described. This paradigm was utilized to investigate effects of protein synthesis inhibiting drugs on learning and retention. Cycloheximide (CXM), which inhibited protein synthesis by over 90% during the training period, did not impair acquisition and did not produce retention deficits an any interval up to 1 day after training. Puromycin (PURO), which inhibited protein synthesis by about 70% during the training period, produced amnesia 5 hr after training, while acquisition was not affected. Thus invertebrates, as well as vertebrates, are susceptible to amnesic effects of puromycin. Although PURO-injected animals showed retention deficits as measured by the number of correct turns, no retention deficit occurred for the behavioral modification consisting of an increase in runway time during the training period. Therefore, PURO appears to show specificity for the different types of longer-term memories that are formed in a training situation.

Animals↗
PubMed1976

Study on cockroach antigen as a probable causative agent in bronchial asthma.

Bronchial provocative test (BPT) with cockroach (CR) antigen was performed in 22 asthmatic subjects. Sixteen had positive reaction (CR-positive) and 6 had negative reaction (CR-negative) to CR antigen by skin test. Immediate bronchoconstrictive response was noted following the antigen inhalation in 14 of 16 CR-positive asthmatics, while none of 6 CR-negative asthmatics showed bronchospasm. Late asthmatic responses also were noted in 13 of 16 CR-positive asthmatic individuals following BPT with CR antigen. The dual asthmatic reactions in CR-positive individuals were mostly inhibited by the prior administration of cromolyn sodium. Three-fold increases in peripheral eosinophil counts were noted 24 hr following BPT with CR antigen. Results indicate that CR-induced asthmatic responses are allergen-specific and CR plays a causative role in allergic asthma in the population studied.

Adolescent↗
PubMed1991

Environmental exposure to cockroach allergens: analysis with monoclonal antibody-based enzyme immunoassays.

Quantitative two-site monoclonal antibody (MAb)-based enzyme-linked immunoassays for two cockroach (CR) allergens, Bla g I and Bla g II, have been developed and used to measure allergen levels in house-dust samples. Dust collected from the CR-infested homes of two patients with asthma from Charlottesville, Va., demonstrated wide variation in the levels of Bla g I, depending on the location of dust collection. Dust from kitchen floors and cabinets contained 50-fold more allergen (mean, 10,755 U/gm of dust) than dust from bedrooms and upholstered furniture (mean, 204 U/gm). One hundred forty-five dust samples were collected from the bedrooms and living rooms of 22 children with asthma and 16 control subjects without asthma living in Atlanta, Ga. Twenty-seven of the 38 homes (17/22 children with asthma; 10/16 control subjects) had detectable Bla g I (4 to 1340 U/gm of dust). Bla g II levels were assayed in 40 kitchen, bedroom, and living room samples from homes in Wilmington, Del. Highest levels of Bla g II were detected in kitchen-floor dust (300 U/gm of dust). Additionally, approximately 20% of homes with no visual evidence of CR infestation had significant levels of Bla g II in at least one dust sample (greater than 4 U/gm of dust). Our results demonstrate that CR may be an occult allergen in homes. The kitchen appears to be the primary site of CR-allergen accumulation, but significant CR-allergen levels can also be found at other sites in the home. The MAb-based assays can be used for quantitation of environmental exposure to CR allergens.(ABSTRACT TRUNCATED AT 250 WORDS)

Allergens↗
PubMed1991

Identification, quantitation, and purification of cockroach allergens using monoclonal antibodies.

A panel of murine IgG monoclonal antibodies (MAbs) was raised against German cockroach (CR) (Blattella germanica) extract and selectively screened to identify MAb directed against allergen(s) recognized by IgE antibodies. Sera from 28 CR-allergic patients were used as sources of IgE antibodies to detect allergens "presented" by the MAb. Four clones (10A6, 3G12, 8F4, and 1D4) produced MAb to allergen(s) that bound IgE antibodies. Quantitative radioimmunoassays were used to compare levels of the MAb-defined allergens in CR extracts. MAb 10A6 reacted with a cross-reacting allergen that was detected in 9/14 CR species, including Blattella, Periplaneta, Blatta, Leucophea, and Supella spp, at concentrations of 100 to 10,000 U/ml. In contrast, MAb 3G12, 8F4, and 1D4 were Blattella specific. The allergen defined by MAb 8F4 was purified by MAb affinity chromatography and size-exclusion by high-performance liquid chromatography. It is a 36 kd heat-sensitive protein, isoelectric point, 5.2 to 5.4. Allergen 10A6 was partially purified by isoelectric focusing and high-performance liquid chromatography. It is a heat-stable, acidic protein (isoelectric point 3.15). Based on comparison of their properties with properties of previously described CR allergens, the allergens defined by MAb 10A6 and 8F4 have been provisionally designated Blattella germanica allergen I (Bla g I) and Blattella germanica allergen II (Bla g II), respectively. Assays of six commercial CR skin test extracts demonstrated a 200-fold difference in Bla g I levels (4.7 to 1085 U/ml) and only two extracts that contained detectable Bla g II (248 and 324 U/ml). The results demonstrate that MAb can be used to identify and define CR allergens and that the strategy of the use of MAb as a first step in allergen analysis and purification can be very effective, especially for poorly characterized allergen extracts.

Allergens↗
PubMed1984

The humoral immune response in the American cockroach, Periplaneta americana: reactivity to a defined antigen from honeybee venom, phospholipase A2.

Our previous experiments demonstrated that honeybee venom could induce a specific, adaptive humoral immune response in the American cockroach. Since honeybee venom is a complex substance made up of several proteins, a more defined antigen is needed for future characterization studies. One of the components of bee venom, phospholipase A2 (PA2) was found to be highly lethal and immunogenic in the roach. Roaches injected with PA2 generated a specific primary response that developed over a period of time, peaking within 10 days, and then gradually subsiding by the fifth week. Specificity of this response was demonstrated by the fact that immunized animals were protected against the original immunizing PA2, but not to PA2 from a heterologous source. In addition, a secondary response could be induced with PA2, demonstrating the existence of immunologic memory. Thus, we established that PA2 could induce as good, if not better, humoral responsiveness as whole bee venom, and therefore could be utilized as a more defined antigen in studies designed to characterize the inducible humoral factor in the roach.

Animals↗
PubMed1985

Preliminary characterization of the inducible humoral factor in the American cockroach (Periplaneta americana).

Previous studies have defined an inducible humoral immune response in the American cockroach that demonstrates specificity and memory. In addition, the response is affected by the age and gender of the animal. Preliminary studies have been initiated to determine which of the protein components found in immune hemolymph might be responsible for mediating this activity. Comparisons between hemolymphs from immune and control animals, using SDS-PAGE gel analysis, revealed that there was a unique band generated in immune animals with an approximate MW of 162K. In addition, several other bands, with MWs of 220K, 115K, 102K, 95K, 80K and 77K, appear to be enhanced in immune hemolymph. The 240K, 220K and 115K bands were found only in females, and so have been temporarily excluded from further study because they were gender-dependent. When hemolymph samples were absorbed with HBT toxoid, the 162K band was no longer present in immune hemolymph, and the 115K, 102K, 95K and the 80/77K doublet were all greatly reduced as compared to controls. Analysis of supernatants from 24 hour cultures of cells from immune and BTS-injected animals, revealed that immune cells secreted the 162K, 102K, 95K and 80K bands. The 115K band was barely detectable, and the 77K band appeared to be missing. Thus, these studies have provided some valuable initial information that reduces the number of proteins that should be considered for further study as mediators of the roach inducible humoral response.

Animals↗
PubMed1985

Effect of gender on the inducible humoral immune response to honeybee venom in the American cockroach (Periplaneta americana).

It is well known that in higher animals, the female of the species usually possesses a superior immune response to that of the male. We investigated the possibility that this rule of nature might also be true amongst the invertebrates. Adult female American cockroaches (Periplaneta americana) were immunized with Honeybee venom, and their responses were compared to that of the adult male. The female primary response was found not only to be enhanced, but prolonged as compared to the male. The response was also specific and demonstrated long-term immunological memory. Thus, the humoral immune response of this advanced invertebrate shares yet another characteristic common to higher vertebrates, since the female of the species demonstrated much better immune responsiveness than the male.

Animals↗
PubMedDate not supplied

Combined intracellular stimulation and high speed video motion analysis of motor control neurons in the cockroach.

A complete understanding of motor control circuitry requires detailed analysis of the behavior produced by the circuitry as well as the connections between individual neurons. A technique is described for combining high speed video motion analysis of leg movements in the cockroach with electrophysiological techniques such as intracellular stimulation/recording from central neurons and EMG recording from leg muscles. Using a restrained preparation, we have quantified leg movements evoked by intracellular stimulation of individual motor neurons and local interneurons. By incorporating motion analysis into the recording paradigm, the transfer functions from electrical activity to movements can be derived. Because distinct and characteristic responses to single and multiple action potentials are seen in slow, intermediate, and fast motor neurons, it is often possible to identify the motor neuron targets of local interneurons. The ability to analyze movement in any plane is especially useful in situations such as blind neuropilar penetrations, where a more restricted motion transducer arrangement may not be in register with the impaled cell. In addition, it is possible to record and analyze such complex phenomena as coordinated movements in multiple joints produced by local interneurons and reflexes produced by proprioceptive feedback due to activity of one motor neuron.

Animals↗
PubMed1983

Functional elimination of afferent pathways and decreased safety factor during postembryonic development of cockroach giant interneurons.

The giant interneurons (GIN) from the cockroach CNS undergo two major physiological changes during the postembryonic developmental period: (A) a marked decrease in the number of afferent pathways innervating the GIN at the metathoracic ganglion (Ts); and (B) a gradual decrease in the safety factor for impulse propagation along the intraganglionic segment in T3. In 100% of the experiments (n greater than 100) performed on GIN from early developmental stages, spontaneous postsynaptic potentials (SPSPs) were recorded; in adults, on the other hand. SPSPs have been recorded in only 34% of the experiments (n = 74). Evoked synaptic potentials can be elicited in nymphal stages by stimulation of 8 nerves of T3, the contralateral connectives, ipsi- and contralateral nerve roots 2, 3, 5, and by stimulation of adjacent GINs. In adult, PSPs can be evoked by stimulation of adjacent GINs, and contralateral thoracic connectives, but not from nerves 2, 3 and 5. The functional disappearance of synaptic inputs to the GINs does not reflect a general phenomenon of reduction in synaptic transmission efficacy. In previous studies it was demonstrated that high frequency stimulation of adult GIN leads to blockage of impulse propagation in T3. In nymphal stages, the safety factor for propagation of impulses along T3 is higher. The reduction in safety factor appears gradually during the postembryonic developmental period. From analysis of the mechanisms underlying the elimination of functional afferent pathways and the appearance of low safety factor (see consecutive paper by Yarom and Spira) it is concluded that the functional elimination of afferents is a consequence of decreased transmission efficacy, while the appearance of low safety regions for impulse propagation is a consequence of morphological changes of the GIN segment within ganglion T3.

Afferent Pathways↗
PubMed1983

Morphological and electrophysiological properties of giant interneurons during the postembryonic development of the cockroach CNS.

The giant interneurons (GIN) of the central nervous system of the cockroach undergo two major physiological changes during the postembryonic development period: (A) a gradual decrease in the safety factor for action potential propagation across the GIN in the metathoracic ganglion (T3); and (B) a marked decrease in the number of afferent pathways innervating the GIN in T3 (Spira and Yarom). Analysis of the morphological structure of the GINs, by intracellular injection of cobalt ions and by cross-sections prepared for light and electron microscope, reveals that despite the extensive growth of the GINs during the postembryonic developmental period, the main structural outline of the fibers is not altered. In adult preparations, however, the GIN diameter narrows 25-26% in ganglion T3, while in early nymphal stages the reduction is only 8-10%. The difference in the extent of narrowing of the fibers in adult and nymphal stages is the major factor that accounts for the development of a low safety factor region for impulse propagation across T3. Analysis of the passive membrane properties of the GIN reveals that the electrotonic length of the GIN segment in T3 is identical in adult and nymphal stages. It is concluded that the functional disappearance of afferents innervating the GINs in T3 is a consequence of decreased transmission efficacy along the afferent pathways.

Afferent Pathways↗
PubMed1983

Double minutes and other chromosomal aberrations in two malignant cell lines of the German cockroach Blattella germanica.

Structural anomalies of mitotic chromosomes from two tumorigenic cell lines of the German cockroach (Blattella germanica) is described. Aberrations, such as unpairable marker chromosomes, double minutes, di- and tricentrics, ring chromosomes, tri- and quadriradials, and chromosome and chromatid gaps and breaks, were observed in varying proportions. This study reports that the double minute chromosomes (DMs) are associated with insect tumor cells, similar to the findings in both murine and human tumor cells.

Animals↗
PubMed1983

Effect of forced locomotion on the rest-activity cycle of the cockroach.

Many insects are known to exhibit clear circadian rest-activity rhythms. Attention has focussed more on active behaviors such as locomotion than on rest. I investigated the consequences of disturbances of the rest period on activity states of the cockroach Leucophea maderae. The 24-h rest-activity rhythm of individual animals maintained under a 12 h light-12 h dark schedule was continuously recorded by timelapse video recording. Three activity states were scored: locomotion, immobility, and limb or antenna movements without locomotion. The effect of (i) 3 h forced activity; and of (ii) handling and a new environment was studied. Forced activity during the last 3 h of the light period caused a significant reduction of locomotion, and of limb or antenna movements; and an enhancement of immobility in the first hours of the dark period. However, over the entire 12 h dark period, these parameters did not differ from control. Brief handling in conjunction with a change in environment during the last 3 h of the dark period, was followed by similar, though smaller changes in locomotion as seen after forced locomotion. The results indicate that the time spent in immobility corresponds to a resting state which is regulated as a function of prior activity. Thus immobility in an invertebrate may exhibit similar regulating mechanisms as rest or sleep in vertebrates.

Animals↗
PubMed1989

A substance resembling somatomedin C in the American cockroach.

Material antigenically resembling somatomedin C (type I insulin-like growth factor, IGF-I) is demonstrated in the American cockroach Periplaneta americana by means of a monoclonal antibody immunoperoxidase technique. It was localized histochemically in neuronal cell somata and axonal fibers (probably interneurons) of the central nervous/neuroendocrine system and in 'endocrine-type' cells lining the midgut epithelium. The IGF-I-like substance is different from vertebrate insulin and also distinct from materials immunostained by different insulin antibodies in the brain and neurohaemal complex of this insect species. These findings are viewed in the light of recent reports on the presence and action of insulin-like chemicals in insects, and with respect to the existence of an insect brain-midgut system similar to the mammalian brain-gastroenteropancreatic system.

Animals↗
PubMedDate not supplied

Structure function analysis of an arthropod peptide hormone: proctolin and synthetic analogues compared on the cockroach hindgut receptor.

Proctolin is a pentapeptide (arg-tyr-leu-pro-thr) found in nervous tissues throughout the phylum Arthropoda. Initially described as a peptidergic neuromuscular transmitter, it now appears that proctolin is a major arthropod neurohormone modulating nervous activity, muscle tonus and contractile force. Structure-function studies with synthetic analogues demonstrate diverse peptides which retain agonistic activity, but few exhibit a high degree of affinity for the cockroach hindgut receptor compared with proctolin (Kdapp = 2 x 10(-8) M). High affinity agonists (Kdapp less than or equal to 10(-7) M) are limited to [phe2]-proctolin, [lys1]-proctolin and specific N-terminal additions. In this regard the hindgut receptor differs in its ligand specificity from that reported for the locust extensor tibia receptor. Using the analogue studies to predict sequences which may act as agonists, we have examined the known vertebrate peptide hormones for proctolin-like sequences. A possible relationship between vasoactive intestinal peptide, proctolin and erythrophore concentrating hormone is critically evaluated.

Amino Acid Sequence↗