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PubMed1984

Voltage-clamp analysis of a calcium-mediated potassium conductance in cockroach (Periplaneta americana) central neurones.

The electrical properties of motoneurone cell bodies in the metathoracic ganglion of the cockroach, Periplaneta americana, have been studied by the voltage-clamp technique. Most experiments were carried out on a single identified cell (cell 28), the cell body of which, as for most other insect motoneurones, is electrically inexcitable. For comparison, some experiments were also carried out on dorsal unpaired median (d.u.m.) cells, the cell bodies of which are excitable. The two cell types differed only in that the d.u.m. cells developed a transient net inward current when depolarized towards zero membrane potential. This current was reduced but not abolished by Ca-free saline. Both cell types had an N-shaped current-voltage relation, as typically seen for molluscan and other neurones, but the location of the falling phase of the relation showed an unusually strong time dependence. The N-shape was abolished by prolonged exposure to Ca-free saline, suggesting it to be due to a K conductance that was activated by the entry of Ca ions through voltage-dependent channels. An outward current was also elicited by ionophoretic injection of Ca ions. The reversal potential of this current varied with the saline K concentration, in the manner expected if the current was carried by K ions. The Ca-mediated K current was blocked by La ions and by the organic Ca antagonist D-600. A series of double-pulse experiments on voltage-clamped cell bodies of cell 28 suggested that very short periods of Ca entry were sufficient to activate the K conductance fully. These experiments also suggested that the K conductance activation was due to intracellular Ca accumulation rather than to its being directly linked to the inward Ca current. The activation of the K conductance by intracellular Ca ions was made more effective by cell membrane depolarization. The Ca-mediated conductance did not inactivate significantly under conditions in which substantial inactivation has been observed in other neurones. The physiological significance of the electrical properties of cell 28 is discussed.

Action Potentials↗
PubMed2004

Fipronil is a potent open channel blocker of glutamate-activated chloride channels in cockroach neurons.

Fipronil, a phenylpyrazole insecticide, displays high insecticidal activity and reduced mammalian toxicity. To better elucidate the mechanism of its selective toxicity between insects and mammals and activity against dieldrin-resistant insects, we studied fipronil action on glutamate-gated chloride channels (GluCls), unique invertebrate ligand-gated chloride channels, in cockroach thoracic ganglion neurons, using the whole-cell patch clamp technique. Glutamate evoked two types of chloride currents, a desensitizing current and a nondesensitizing current. Fipronil differentially inhibited these two types of currents with different potencies and with different rates of reversibility. Fipronil inhibited the desensitizing and nondesensitizing GluCls with IC50 values of 801 and 10 nM, respectively. Kinetic analysis revealed that fipronil blocks required channel opening. Recovery of the desensitizing current from fipronil block required channel opening, whereas recovery of nondesensitizing current from block was independent of channel opening. The high potency of fipronil against the nondesensitizing current was due to a slow unblocking rate constant. In addition, when the nondesensitizing GluCls were occupied by picrotoxinin, the receptors became less sensitive to fipronil block. It is concluded that GluCls are a critical target for fipronil, especially for the selective toxicity between mammals and insects, and that fipronil block of GluCls may play a role in the lack of the cross-resistance with dieldrin.

Animals↗
PubMed1963

CARBOHYDRATE DIGESTION IN THE COCKROACH.

The digestive tract of the cockroach, Blaberus craniifer Burmeister, contains amylase, alpha- and beta-glucosidases, beta-fructofuranosidase, alpha- and beta-galactosidases, but no cellulase. The most active site of hydrolytic action is the midgut, followed by the foregut, hindgut, salivary gland tissue, and Malpighian tubules.

Animals↗
PubMed1965

BLOOD OF A COCKROACH: UNUSUAL CELLULAR BEHAVIOR.

In blood smears of a cockroach a small cell occurs which is frequently paired with a larger anucleate cytoplasmic body. The larger body in such pairs is free of nucleic acid, but contains polysaccharides, as demonstrated by the periodic acid-Schiff reaction. At one extreme of a series of these associations, the cell is completely distinct from the cytoplasmic body. At the other extreme the nucleus of the cell is within and apparently a part of the larger body. A graded series of pairs suggests a mechanism resembling phagocytosis in which the cytoplasmic body either gains or regains cell status by retaining the nucleus of the ingested member.

Animals↗
PubMed1978

Effect of metronidazole on the intestinal microflora of the american cockroach, Periplaneta americana l.

The effect of feeding to cockroaches the antimicrobial agent metronidazole, which acts specifically against anaerobes, was assessed by light and scanning electron microscopy, bacteriological examination, and methane formation. Types of organisms and total numbers were greatly reduced from controls. The health of mature adults was unaffected, but stunting occurred in subadult animals maintained on the antibiotic from hatching. The intracellular bacteria of the fat body were not affected by the drug. The results are discussed with respect to a proposed microbe-host extracellular symbiosis.

Animals↗
PubMed1979

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.

Animals↗
PubMed1975

Ultrastructal morphology of some prokaryotic microorganisms associated with the hindgut of cockroaches.

Scanning electron microscopy and transmission electron microscopy have been used to visualize the morphology and ultrastructure of two types of microorganisms in the hindgut of the cockroach Blaberus posticus. Both organisms, designated as either short or long rods, are attached to chitinous projections from the gut wall. Micrographs suggest that the organisms are prokaryotic with a cell wall complex characteristic of gram-negative bacteria. However, certain differences were noted between the cell wall complex of the two types. Two forms of the long-rod type were noted, with one form appearing to be a "degenerate" or "transitional" cell. In the degenerate cells, vesicles are observed that often are contiguous with the cytoplasmic membrane. There are indications that the long-rod type may divide by longitudinal fission. Neither the short- nor long-rod type has been cultivated in its respective recognizable form.

Animals↗
PubMed1979

Novel microbial and chemical components of a specific black-band region in the cockroach hindgut.

An area of the hindgut of the cockroach, Eublaberus posticus, is characterized by its black color. This area is the site of accumulation of metal sulfides in the lumen next to the gut wall. In addition to the normal hindgut flora, two unusual procaryotic organisms are seen by both scanning and transmission electron microscopy only in this area of the hindgut. They are (i) a large rod (1.2 by 6 micrometers) with a tuft of polar flagella, many inclusion bodies, and a distinctive complex wall and (ii) an apparently flexible rod with a helically ridged wall. In addition, phagelike particles are described which are apparently infecting gram-positive bacteria attached to the gut wall in the black band area.

Animals↗
PubMed1984

Pleomorphism of fusobacteria isolated from the cockroach hindgut.

Fusobacteria are commonly isolated from the hindgut of the cockroach Eublaberus posticus . Eleven strains isolated from E. posticus by us were keyed to four species, Fusobacterium necrophorum, F. varium , F. gonidiaformans , and F. prausnitzii , using current taxonomic criteria. With the exception of F. gonidiaformis , all species showed rods with swollen centers and large bodies. The pleomorphism of F. varium was examined by phase microscopy and scanning and transmission electron microscopy. The pleomorphic process begins with a gradual swelling at the center of the rod until a large round body is formed. Some of these round bodies then fragment, giving rise to rod-shaped cells. When 10% yeast extract was added to growth media, pleomorphism was not induced. A dialyzable factor was found to account for this observation. Fermentation of [1-14C]glutamic acid gives rise to butyrate labeled in the carboxyl carbon, indicating that butyrate is formed by the hydroxyglutarate pathway which may be characteristic for the genus Fusobacterium.

Animals↗
PubMed1983

The possible role of juvenile hormone esterase in the regulation of juvenile hormone titre in the female cockroach Diploptera punctata.

Juvenile hormone (JH) degradation in vitro and in vivo was studied in the viviparous cockroach Diploptera punctata. In vitro studies with juvenile hormone esterase (JHE) and "general" or l-naphthyl acetate esterase (NAcE) revealed that diethyl-p-nitrophenylphosphate (paraoxon) inhibited both JHE and NAcE activity, but the latter was more sensitive and was completely inhibited at 0.1 mM. NAcE activity was resistant to inhibition with Triton X-100, whereas JHE activity in haemolymph of adult females was inhibited 100% at Triton X-100 concentration of 0.25%. Eighty percent inhibition of JHE activity in vivo was observed following injection of 0.2 microL Triton X-100. In contrast to the previously observed dose-dependant increase in JHE activity. NAcE activity did not increase following treatment of allatectomized females with the JH analogue ethyl-2E,4E-3,7,11-trimethyl-2,4-dodecadienoate (hydroprene). Hydroprene was not catabolized in haemolymph of D. punctata in vitro. The half-life of C16 JH (JH III) in the haemolymph, in vivo, was 1.65 h for day 5 females and 2 h for day 6 females.

Animals↗
PubMed1992

A student apparatus for recording action potentials in cockroach legs.

A comparatively simple apparatus allows even beginning students to observe action potentials in the cockroach leg. The recordings are made extracellularly by impaling the leg on two insect pins. Deflection of large spines on the leg, which are each innervated by one sensory neuron, initiates the action potentials. Using this technique, students observe the all-or-nothing nature of action potentials, their coding of information by frequency, and sensory adaptation.

Action Potentials↗
PubMed1983

Ionic distribution in dopamine-stimulated NaCl fluid-secreting cockroach salivary glands.

The compound, racemose, innervated salivary glands of the cockroach Periplaneta americana closely resemble in structure and function the mammalian salivary glands (C.R. House, Biol. Rev. 55: 417-473, 1980). The quantitative distribution of Na, K, Cl, Ca, Mg, P, and S was investigated in the P-cells secreting isotonic NaCl and in duct lumens by using electron probe X-ray microanalysis of frozen-hydrated and frozen-dried cryosections. In the unstimulated glands in vitro the cells had (mmol/l cell-H2O) Na, 10; K, 110; Cl, 39 while the primary saliva had Na, 153; K, 4; Cl, 151. With 1 mumol dopamine in the bathing Ringer solution the P-cells had Na, 25; K, 177; Cl, 58 and the primary saliva, Na, 153; K, 26; Cl, 172. During passage through the ducts, the primary saliva was modified by an absorption of NaCl: more in unstimulated than in stimulated glands. It is proposed that the cells have a Na-K-ATPase both in the apical and basal cell membrane, as in vertebrate choroid plexus, and dopamine might increase the K and Na conductance of the basal cell membrane.

Animals↗
PubMed1975

Nonspiking interneurons in walking system of the cockroach.

Intracellular recordings were made from the neurites of interneurons and motoneurons in the metathoracic ganglion of the cockroach, Periplaneta americana. Many neurons were penetrated which failed to produce action potentials on the application of large depolarizing currents. Nevertheless, some of them strongly excited and/or inhibited slow motoneurons innervating leg musculature, even with weak depolariziing musculature, even with weak depolarizing currents. Cobalt-sulfide-straining of these nonspiking neurons showed them to be interneurons with their neurites contained entirely within the metathoracic ganglion. Two further characteristics of these interneurons were rapid spontaneous fluctuations in membrane potential and a low resting membrane potential. One nonspiking neuron, interneuron I, when depolarized caused a strong excitation of the set of slow levator motoneurons which discharge in bursts during stepping movements of the metathoracic leg. During rhythmic leg movements the membrane potential of interneuron I oscillated with the depolarizing phases occurring at the same time as bursts of activity in the levator motorneurons. No spiking or any other nonspiking neuron was penetrated which could excite these levator motoneurons. From all these observations we conclude that oscillations in the membrane potential of interneuron I are entirely responsible for producing the levator bursts, and thus for producing stepping movements in a walking animal. During rhythmic leg movements, bursts of activity in levator and depressor motoneurons are initiated by slow graded depolarizations. The similarity of the synaptic activity in these two types of motoneurons suggests that burst activity in the depressor motoneurons is also produced by rhythmic activity in nonspiking interneurons. The fact that no spiking neuron was found to excite the depressor motoneurons supports this conclusion. Interneuron I is also an element of the rhythm-generating system, since short depolarizing pulses applied to it during rhythmic activity could reset the thythm. Long-duration current pulses applied to interneuron I in a quiescent animal did not produce rhythmic activity. This observation, together with the finding that during rhythmic activity the slow depolarizations in interneuron I are usually terminated by IPSPs, suggests that interneuron I alone does not generate the rhythm. No spiking interneurons have yet been enccountered which influence the activity in levator motoneurons. Thus, we conclude that the rhythm is generated in a network of nonspiking interneurons. The cellular mechanisms for generating the oscillations in this network are unknown. Continued.

Action Potentials↗
PubMed1976

Ultrastructure of region of a low safety factor in inhomogeneous giant axon of the cockroach.

1. The structure of the ventral giant axons of the cockroach at the level of ganglion T3 was studied by means of light and electron microscopy. 2. From serial sections and cobalt injections, the axons diameter was found to range between 40 and 60 mum at the caudal end of ganglion T3; toward the center of T3 they narrow to 20-40 mum, and again expand to 30-45 mum anteriorly in ganglion T3. 3. Each giant axon sends off several branches, 1-15 mum in diameter, into the neuropil. The giant axons and the bases of their branches are enveloped by cytoplasmic processes of glial cells. The periaxonal space is about 100-200 A. 4. Distally the branches are devoid of glial envelopes and the extracellular space between the branches and other axonal profiles is about 200 A. Terminals with presumptive chemical synapses on the giant axon branches were found. Clear vesicles, 300-400 A in diameter, are seen clustered together. The width of the supposedly synaptic gap is about 100 A. 5. In some areas the branches and other axonal profiles form close appositions.

Animals↗
PubMed1989

Two components of rapid sensory adaptation in a cockroach mechanoreceptor neuron.

1. The femoral tactile spine of the cockroach is innervated by a single sensory neuron with its cell body in the spine. A step deflection of the spine produces a burst of action potentials which decays to zero in approximately 1 s. This rapid adaptation occurs during encoding of action potentials and can be studied by extracellular electrical stimulation of the sensory neuron. 2. The threshold current of the neuron is labile, increasing with depolarization and decreasing with mild hyperpolarization. During rapid adaptation, the threshold current increases to exceed any steady stimulating current. 3. The dynamic behavior of the threshold current in the tactile spine neuron was observed following step changes in membrane current. The threshold current followed a trajectory which could always be well fitted by a sum of two exponential decays with time constants of approximately 100 and 1,000 ms. 4. The amplitude of the slow component of threshold change was proportional to the size of the step change in membrane current but saturated at strong hyperpolarizations. Its time constant decreased monotonically with depolarization. In contrast, the fast component had more complex behavior, changing biphasically with step amplitude and reversing with initial hyperpolarizations. Its time constant was maximal at the resting membrane potential. 5. The amplitude of the fast component was strongly reduced by treatment with N-chlorosuccinimide (NCS), an agent which reduces sodium channel inactivation. It was not affected by some agents known to block potassium channels. 6. The slow component was not affected by NCS, tetraethyl-ammonium chloride, apamin, or charybdotoxin. 7. Approximate contributions of the two components to the steady-state threshold were reconstructed from step responses. These indicate that the slow component dominates when the receptor is hyperpolarized. However, during depolarizations from rest, as occur with normal stimulation, the two components contribute approximately equally to threshold changes.

Adaptation, Physiological↗
PubMed1995

Competitive interactions between supernumerary and normal sensory neurons in the cockroach are mediated through a change in quantal content and not quantal size.

1. The final steps in synapse formation and stabilization involve the adjustment of strength of connections through competitive interactions between neurons contacting a common target. The mechanisms underlying this competition-driven adjustment of synaptic strength are not well understood. We have studied one aspect of this phenomenon using the cercal sensory system of first instar cockroach nymphs. 2. Quantal analysis of excitatory postsynaptic potentials (EPSPs) recorded at the synapse between the lateral filiform hair sensory neuron (L) and giant interneuron 3 (GI3) was carried out to determine whether the reduction in EPSP amplitude observed in the presence of an additional sensory neuron ("space invader neuron" or SIN) was due to pre- or postsynaptic changes. 3. Mean quantal content, not quantal size, was reduced at the L-GI3 synapse in the presence of a SIN. Fitting binomial distributions to the EPSP amplitude histograms gave estimates of the number of releasable quanta or of synaptic sites (n) and the probability of release at these sites (p). The reduction in quantal content is associated with a decrease in the binomial parameter n and not p, suggesting that there is a decrease in the number of contacts, release sites, or quanta available for release, rather than a change in probability of release.

Animals↗
PubMed1996

Two distinct low-voltage-activated Ca2+ currents contribute to the pacemaker mechanism in cockroach dorsal unpaired median neurons.

1. The contribution of Ca2+ currents to the endogenous firing properties of cockroach isolated adult dorsal unpaired median neurons was investigated using whole cell patch-clamp technique with 5 mM Ca2+ as the charge carrier. At least three types of Ca2+ currents, a high-voltage-activated Ca2+ current and two low-voltage-activated (LVA) Ca2+ currents, have been found in these neurons. This study focused on the LVA Ca2+ currents, which are suitable candidates in the generation of the slow predepolarization because of their low threshold of activation. 2. The global LVA Ca2+ current could be dissociated by means of nickel sensitivity, deactivation time constant and voltage dependence of time to peak, tail current amplitude and inactivation, as transient and maintained LVA Ca2+ currents. 3. The transient LVA Ca2+ current, sensitive to 100 microM Ni2+, was isolated by using a subtraction procedure. It was activated at -70 mV and half-inactivated at -59.5 mV. The inactivation was purely voltage dependent. Current-clamp experiments performed with 150 microM Ni2+ indicated that this current was involved in the initial part of the predepolarization. 4. The maintained LVA Ca2+ current, resistant to 100 microM Ni2+, was activated in a range of potential 10 mV more positive than the transient LVA Ca2+ current, and its voltage dependence of inactivation displayed a U-shaped-curve. 5. Replacing Ca2+ with Ba2+ in equimolar amount or low internal Ca2+ concentration [5 mM bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) in the pipette] induced a monotonic voltage dependence of inactivation and increased the rate of relaxation of this current. These effects were mimicked by high internal Ca2+ concentration [0.1 mM Ca2+ and no ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid in the pipette]. This demonstrated an unusual Ca2+-sensitive inactivation process that varied over a narrow range of Ca2+ concentrations. 6. Current-clamp experiments performed under 150 microM Ni2+, with 15 mM external Ca2+ concentration (which potentiated the maintained LVA current within 30 s of superfusion) or with 5 mM BAPTA in the pipette demonstrated the participation of this current in the last two-thirds of the slow predepolarizing phase. 7. Our findings demonstrated, for the first time in neurosecretory cells, the coexistence of two distinct LVA Ca2+ currents, which have specialized function in the generation of the pacemaker activity.

Animals↗