Variations of glycogen: II. Following deafferentation of Knollenorgan sensory cells,a lateral line electroreceptor of mormyrid fish

The effect of deafferentation on glycogen metabolism was studied in the sensory cells of mormyrid Knollenorgan electroreceptors. Glycogen was visualized in the sensory cells after fixation in a solution containing potassium ferricyanide and osmium tetroxide. The density variations of glycogen were evaluated by a morphometric method. Sectioning of the afferent nerve results in a cessation of the spontaneous receptor cells activity after 48 h and the glycogen content of these cells increases three fold in the first 5 days after nerve transection. From day 5 on, the glycogen concentration diminishes progressively until day 13. After the sensory cells had become completely deformed, the quantification of glycogen particles was no longer possible and the degeneration of the sensory cells was complete within 20 days after nerve section. These results show that (1) the afferent nerve fibre is indispensable for the anatomo-functional maintenance of the sensory cells and (2) the nerve has only an indirect influence on glycogen variations within the sensory cells.     

Energy utilization and gluconeogenesis in isolated leech segmental ganglia: Quantitative studies on the control and cellular localization of endogenous glycogen

The isolated segmental ganglia of the horse leech Haemopis sanguisuga were used as a model system to study the utilization and control of glycogen stores within nervous tissue. The glycogen in the ganglia was extracted and assayed fluorimentrically and its cellular localization and turnover studied by autoradiography in conjunction with [³H]glucose. We measured the glycogen after various periods of electrical stimulation and after incubation with K⁺, Ca²⁺, ouabain and glucose. The results for each experimental ganglion were compared to a paired control ganglion and the results analysed by paired t-tests. Electrical stimulation caused sequential changes in glycogen levels: a reduction of up to 67% (5–10 min); followed by an increase of up to 124% (between 15–50 min); followed by a reduction of up to 63% (60–90 min). Values were calculated for glucose utilization (e.g. 0.53 μmol glucose/gm wet weight/min after 90 min) and estimates derived for glucose consumption per action potential per neuron (e.g. 0.12 fmol at 90 min). Glucose (1.5–10 mM) increased the amount of glycogen (1.5 mM by 30% at 60 min) and attenuated the effects of electrical stimulation. Ouabain (1 mM) blocked the effect of 5 min electrical stimulation. Nine millimolar K⁺ increased glycogen by 27% after 10 min and decreased glycogen by 34% after 60 min; 3 mM Ca²⁺ had no effect after 10 or 20 min and decreased glycogen by 29% after 60 min. Other concentrations of K⁺ and Ca²⁺ reduced glycogen after 60 min. Autoradiographic analysis demonstrated that the effects of elevated K⁺ were principally within the glial cells. We conclude that (i) the glycogen stores in the glial cells of leech segmental ganglia provide an endogenous energy source which can support sustained neuronal activity, (ii) both electrical stimulation and elevated K⁺ can induce gluconeogenesis within the ganglia, (iii) that electrical activation of neurons produces changes in the glycogen in the glial cells which are controlled in part by changes in K⁺.     

Sensory interaction with central ‘generators’ during respiration in the dogfish

Summary The activity in sensory and motor nerves of the gills was recorded from selected branches of the vagus nerve in decerebrate dogfish,Scyliorhinus canicula. Vagal motoneuronal activity was observed at the start of the rapid pharyngeal contraction and was followed by sensory nerve activity which preceded the slow expansion phase. Rhythmical vagal motoneuronal activity was still present after all movements had been prevented by curare paralysis although the frequency of the rhythm was higher than in the ventilating fish. Electrical stimulation of vagal sensory fibres had 3 effects on the ventilatory movements. (1) It evoked a reflex contraction of several gill muscles after a latency of about 11 ms. (2) It could reset the respiratory cycle because a stimulus given during expansion delayed the onset of the subsequent contraction. (3) The stimulus could entrain the rhythm if it was given continuously at a frequency close to that of ventilation. The vagal motor rhythm was disrupted by trigeminal nerve stimulation in the paralyzed fish but not if the motor rhythm was being entrained by vagal nerve stimulation. Vagal sensory activity may be important, therefore, in maintaining the stability of the generating circuits.Abbreviation LED Light emitting diode     

Mechanism of glycogen supercompensation in rat skeletal muscle cultures

A model to study glycogen supercompensation (the significant increase in glycogen content above basal level) in primary rat skeletal muscle culture was established. Glycogen was completely depleted in differentiated myotubes by 2 h of electrical stimulation or exposure to hypoxia during incubation in medium devoid of glucose. Thereafter, cells were incubated in medium containing glucose, and glycogen supercompensation was clearly observed in treated myotubes after 72 h. Peak glycogen levels were obtained after 120 h, averaging 2.5 and 4 fold above control values in the stimulated- and hypoxia-treated cells, respectively. Glycogen synthase activity increased and phosphorylase activity decreased continuously during 120 h of recovery in the treated cells. Rates of 2-deoxyglucose uptake were significantly elevated in the treated cells at 96 and 120 h, averaging 1.4–2 fold above control values. Glycogenin content increased slightly in the treated cells after 48 h (1.2 fold vs. control) and then increased considerably, achieving peak values after 120 h (2 fold vs. control). The results demonstrate two phases of glycogen supercompensation: the first phase depends primarily on activation of glycogen synthase and inactivation of phosphorylase; the second phase includes increases in glucose uptake and glycogenin level.     

Changes in the content of glycogen and its metabolites during acute exposure ofAnabas testudineus (Bloch) to furadan

Significant differences were observed in glycogen metabolism ofAnabas testudineus exposed to an acute lethal (1.56 mg/litre) and a sublethal (0.56 mg/litre) concentration of furadan. At sublethal concentration, the muscle glycogen which was utilized during the early periods of exposure, was replenished in the later period of exposure and at 120 h, the muscle glycogen levels were higher than the control. At higher concentration, the liver glycogen levels showed an increase presumably at the expense of fuel reserves of the muscle.     

Metabolic responses to exhaustive exercise change markedly during the protracted non-trophic spawning migration of the lamprey <Emphasis Type="Italic">Geotria australis</Emphasis>

Adults of the Southern hemisphere lamprey Geotria australis were subjected to an exercise/recovery regime at the commencement and end of their 12–15 month non-trophic, upstream spawning migration. In early (immature) migrants and pre-spawning females, muscle glycogen was markedly depleted during exercise, but became rapidly replenished. As muscle lactate rose during exercise and peaked 1–1.5 h into the recovery period, and therefore after muscle glycogen had become replenished, it cannot be the direct source for that replenishment. However, both plasma lactate and glycerol (but not muscle glycerol and glucose) rose sharply during exercise and then declined markedly during the first 0.5 h of recovery and thus exhibited the opposite trend to that of muscle glycogen, implying that these limited pools of glycogenic precursors contribute to glycogen replenishment. Although plasma glucose rose following exercise, and consequently could also be a precursor for muscle glycogen replenishment, it remained elevated even after muscle glycogen had become replenished. While resting pre-spawning females and mature males retained high muscle glycogen concentrations, this energy store became permanently depleted in females during spawning. In mature males, muscle glycogen remained high and lactate low during the exercise/recovery regime, whereas muscle glycerol declined precipitously during exercise and then rose rapidly. In summary, vigorous activity by G. australis is fuelled extensively by anaerobic metabolism of glycogen early in the spawning run and by pre-spawning females, but by aerobic metabolism of its energy reserves in mature males.     

Modulation of auditory responsiveness in the locust

The auditory responsiveness of a number of neurones in the meso- and metathoracic ganglia of the locust, Locusta migratoria, was found to change systematically during concomitant wind stimulation. Changes in responsiveness were of three kinds: a suppression of the response to low frequency sound (5 kHz), but an unchanged or increased response to high frequency (12 kHz) sound; an increased response to all sound; a decrease in the excitatory, and an increase in the inhibitory, components of a response to sound. Suppression of the response to low frequency sound was mediated by wind, rather than by the flight motor. Wind stimulation caused an increase in membrane conductance and concomitant depolarization in recorded neurones. Wind stimulation potentiated the spike response to a given depolarizing current, and the spike response to a high frequency sound, by about the same amount. The strongest wind-related input to interneuron 714 was via the metathoracic N6, which carries the axons of auditory receptors from the ear. The EPSP evoked in central neurones by electrical stimulation of metathoracic N6 was suppressed by wind stimulation, and by low frequency (5 kHz), but not high frequency (10 kHz), sound. This suppression disappeared when N6 was cut distally to the stimulating electrodes. Responses to low frequency (5 kHz), rather than high frequency (12 kHz), sounds could be suppressed by a second low frequency tone with an intensity above 50-55 dB SPL for a 5 kHz suppressing tone. Suppression of the electrically-evoked EPSP in neurone 714 was greatest at those sound frequencies represented maximally in the spectrum of the locust's wingbeat. It is concluded that the acoustic components of a wind stimulus are able to mediate both inhibition and excitation in the auditory pathway. By suppressing the responses to low frequency sounds, wind stimulation would effectively shift the frequency-response characteristics of central auditory neurones during flight.     

c-fos Expression in Gracilothalamic Tract Neurons after Electrical Stimulation of the Injured Sciatic Nerve in the Adult Rat

The number of c-fos protein-like immunoreactive (Fos-LI) cells in the gracile nucleus was determined after electrical stimulation at Aα/Aβ-fiber strength of the normal and of the previously injured sciatic nerve in adult rats. No Fos-LI cells were seen after electrical stimulation of the noninjured sciatic nerve, or after sciatic nerve injury without electrical stimulation. However, stimulation 21 days after sciatic nerve transection resulted in numerous Fos-LI cells in the ipsilateral gracile nucleus. Combined Fos immunocytochemistry and retrograde labeling from the thalamus showed that the majority (76%; range = 70–80%) of the cells in the gracile nucleus that expressed Fos-LI after nerve injury projected to the thalamus. The results indicate that morphological, biochemical, and physiological alterations in primary sensory central endings and second-order neurons, which have earlier been demonstrated in the dorsal column nuclei after peripheral nerve injury, are accompanied by changes in the c-fos gene activation pattern after stimulation of the injured sciatic nerve. A substantial number of the c-fos-expressing neurons project to the thalamus.     

Convergence of mechanosensory inputs onto neuromodulatory serotonergic neurons in the leech

By the frequency-dependent release of serotonin, Retzius neurons in the leech modulate diverse behavioral responses of the animal. However, little is known about how their firing pattern is produced. Here we have analyzed the effects of mechanical stimulation of the skin and intracellular stimulation of mechanosensory neurons on the electrical activity of Retzius neurons. We recorded the electrical activity of neurons in ganglia attached to their corresponding skin segment by segmental nerve roots, or in isolated ganglia. Mechanosensory stimulation of the skin induced excitatory synaptic potentials (EPSPs) and action potentials in both Retzius neurons in a ganglion. The frequency and duration of responses depended on the strength and duration of the skin stimulation. Retzius cells responded after T and P cells, but before N cells, and their sustained responses correlated with the activity of P cells. Trains of five impulses at 10 Hz in every individual T, P, or N cell in isolated ganglia produced EPSPs and action potentials in Retzius neurons. Responses to T cell stimulation appeared after the first impulse. In contrast, the responses to P or N cell stimulation appeared after two or more presynaptic impulses and facilitated afterward. The polysynaptic nature of all the synaptic inputs was shown by blocking them with a high calcium/magnesium external solution. The rise time distribution of EPSPs produced by the different mechanosensory neurons suggested that several interneurons participate in this pathway. Our results suggest that sensory stimulation provides a mechanism for regulating serotonin-mediated modulation in the leech.     

Design and performance of an electrical stimulator for long-term contraction of cultured muscle cells

Excitability in muscle cells manifests itself as contractility and may be evoked by electrical stimulation. Here we describe an electrical stimulator device applicable to cells seeded on standard multiwell plates and demonstrate how it effectively stimulates synchronous contraction of skeletal muscle C2C12 cells without damaging them. The electrical stimulator of cultured cells (ESCC) consists of two connection cards and a network of platinum electrodes positioned in such way that each well in a row is uniformly stimulated. The ESCC may produce a range of outputs based on the stimulation parameters it receives from a commercial pulse generator and can be placed in a standard cell incubator, allowing for long-term stimulation as required for biochemical and molecular biological assays. We show that a 90-min stimulation of C2C12 myotubes at 50 V, 30 ms of pulse duration, and 3 Hz of frequency enhances glucose metabolism and glycogen mobilization while oppositely modulating the activity ratio of glycogen metabolizing enzymes. Thus, we demonstrate that long-term electrical stimulation of C2C12 myotubes with the ESCC results in contractility and metabolic changes, as seen in exercising muscle.     

Functional organization of the tympanal organ of the flour moth, Ephestia kuehniella

Data about electrical recordings from the tympanic organ of the flour moth, Ephestia kuehniella, to acoustic stimuli is presented. The stimuli had a frequency that ranged from 5 to 100 kHz, with minimal intensities of 40 to 50 db (Odb = 0.0002 dynes/cm²) and maximal up to 110 db. The tympanic organ of E. kuehniella responded in the whole range of frequencies used and showed two sensitivity maxima, one at 20 kHz and the other at 60 kHz. It responded from 45 to 110 db. The electrical activity of the tympanic nerve consisted of a spontaneous discharge of a type B cell and a tonic discharge in response to acoustic stimulation, produced by four acoustic sense cells, called A₁, A₂, A₃, and A₄. All these acoustic sense cells respond in the whole frequency range used and they differ in the heights of their action potentials and in their sensitivity to acoustic stimuli. The possible biological significance of hearing in pyralid moths is discussed.     

Effect of reticular formation on hippocampal neurons (field CA1)

The reaction of field CA₁ hippocampal neurons to stimulation of the reticular formation (RF) with impulses of different frequencies was investigated in experiments on unanesthetized rabbits. The effect of electrical and sensory stimuli was compared and the effect of reticular stimulation on the sensory responses was determined. With an increase in the frequency of RF stimulation, the number of neurons of field CA₁ responding with inhibition of the activity increases. Multimodal neurons of the hippocampus depend on the reticular input to a greater degree than unimodal neurons. Neurons whose activity does not change in response to the effect of sensory stimuli also do not respond to stimulation of the RF. Neurons responding with inhibitory reactions to sensory stimulation show a higher correlation with the effects of RF stimulation than neurons with activation reactions and, especially those with "complex" responses to the effect of sensory stimuli. In a considerable number of hippocampal neurons the responses to sensory stimuli change in the course of 10–15 min after stimulation of the RF. The role of the RF in the organization of the reactions of hippocampal neurons is discussed.Division of Memory Problems, Institute of Biological Physics, Academy of Sciences of the USSR, Pushchino-on-Oke. Translated from Neirofiziologiya, Vol. 3, No. 3, pp. 227–235, May–June, 1971.     

Fat and glycogen utilization in the larynx muscles of fire-bellied toads (Bombina bombina L.) during calling activity

Summary Both fat and glycogen are present in abundance in the larynx muscles of anurans. To clarify their role, the Musculus dilatator laryngis of the male fire-bellied toad, Bombina bombina was studied.In untreated males, the mean fat content of this larynx muscle was 14%; the muscle contained glycogen amounting to 57% of that measured in the liver tissue of the same animal. After thirteen days of continual calling activity induced by administration of a gonadotropin, the fat content fell to 6%, the glycogen to 34%. The fat content was essentially unchanged (13%) by four hours of electrical stimulation of the muscle; the glycogen content, however, had fallen to 42% after this treatment. Neither component was altered in amount by castration, the fat content being about 13% and that of glycogen, 52%. Nor did treatment with gonadotropic hormone reduce either the fat content (13%) or the amount of glycogen (59%).From these results it was concluded that fats represent a direct source of energy for the larynx muscles, which is used up gradually over long periods of calling. The glycogen in these muscles, on the other hand, is a short-term store sufficient to supply energy for only a few hours of calling activity.Supported by the Deutsche Forschungsgemeinschaft.     

Evoked frequency stabilization in the electrical activity of the cat brain

In this study, our previous results on the important relation between EEG and EPs were extended by experiments with chronically implanted and freely moving cats, which had electrodes at the acoustical cortex, inferior colliculus and reticular formation. During the experiments the frequency stabilization upon sound stimulation was shown in the frequency domain by comparison of the pre-stimulus power spectra and post-stimulus amplitude frequency characteristics. Comparative frequency domain analysis of about 75 EEG-EPograms (sample of spontaneous activities just prior to stimulation and single evoked potentials following the stimulation), which were recorded from all the brain nuclei mentioned above and from each of the 11 cats, was performed as follows: 1) Power spectra of the EEG-records prior to stimulus were evaluated. 2) Instantaneous frequency characteristics of single EPs were obtained by the Fourier transform. 3) Distribution of the amplitude maxima of the EP-frequency characteristics and the distribution of the EEG-spectral peaks were compared by plotting two types of histograms containing relevant spectral peaks before and after the stimulation. In a frequency range between 1–1000 Hz, the frequency distribution of the EP records from RF and IC were accumulated in narrow discrete frequency channels, whereas, the distribution of the spectral peaks of the EEG depicted frequency spread in broad channels. The frequency stabilization of the EP records from GEA, in the alpha frequency range, was also observed. This effect was described by a factor which we called as the Frequency Stabilization Factor. The results presented in this study showed that the frequency stabilization of the brain's electrical activity induced by sensory stimulation displayed a fluctuation leading to frequency stabilization factors between 0.95 and 5.00. The frequency stabilization and relevant power enhancement upon stimulation strongly support our contention that evoked potential results from the frequency stabilization of the spontaneous activity, triggered by stimulation.Supported by Grant No. TAG-345 of the Scientific and Technical Research Council of Turkey     

Evidence of a neural loop involved in controlling spermathecal contractions in Locusta migratoria

The control of spermathecal contractions in Locusta migratoria via a neural loop was demonstrated using mechanical stimulation and electrophysiological recordings. Extracellular electrophysiological recordings from the receptaculum seminis nerve (N2B2), which innervates the spermathecal sac, were conducted during mechanical stimulation of the genital chamber sensory cells. Activation of the genital chamber sensory cells, using a glass probe approximating the shape and size of an egg, was found to increase the action potential frequency and initiate bursts of action potentials if a tonic frequency of action potentials was present prior to stimulation. If the motor pattern initially consisted of bursts of action potentials, then mechanical stimulation of the genital chamber sensory cells resulted in an increase in firing frequency, in most preparations, with the bursting remaining. Removal of the probe from the genital chamber always returned the motor activity to that noted prior to sensory cell stimulation. Simultaneous electrophysiological recordings from both the left and right receptaculum seminis nerves (N2B2) revealed that the bursts of action potentials were coordinated, although individual action potentials were not coupled one to one. Activation of the genital chamber sensory cells also resulted in increases in spermathecal contraction frequency, an effect which was coordinated with the changes in motor activity. It is proposed that an egg in the genital chamber activates the sensory cells resulting in increases in spermathecal contraction frequency and the subsequent release of spermatozoa onto the micropyle of the egg for fertilization.     

Mechanism of conduction block in amphibian myelinated axon induced by biphasic electrical current at ultra-high frequency

The mechanism of axonal conduction block induced by ultra-high frequency (≥20 kHz) biphasic electrical current was investigated using a lumped circuit model of the amphibian myelinated axon based on Frankenhaeuser-Huxley (FH) equations. The ultra-high frequency stimulation produces constant activation of both sodium and potassium channels at the axonal node under the block electrode causing the axonal conduction block. This blocking mechanism is different from the mechanism when the stimulation frequency is between 4 kHz and 10 kHz, where only the potassium channel is constantly activated. The minimal stimulation intensity required to induce a conduction block increases as the stimulation frequency increases. The results from this simulation study are useful to guide future animal experiments to reveal the different mechanisms underlying nerve conduction block induced by high-frequency biphasic electrical current.     

Comparative characteristics of septal "burst" neurons after elimination of afferent effects of the reticular formation in the rabbit

Comparative analysis of characteristics of rhythmic theta-activity in the neurones of the medial septal nucleus and nucleus of diagonal band was performed in intact rabbits after. i. v. injection of pentobarbital, and in rabbits with chronic lesion of the ascending brain-stem afferent fibers. In both conditions theta-bursts disappeared in some cells with unstable periodic rhythmic modulation; substantial population of the septal units preserved regular burst activity. Main characteristics of theta-bursts were almost identical in both states, their mean frequency decreased to 3.5 Hz. The theta-rhythm in hippocampal EEG was usually absent; but low-frequency rhythmic activity could be evoked by electrical or sensory stimulation as well as by injection of bemegrid or physostigmine. The data show that the ascending brain-stem afferents control: the frequency of the bursts in a population of septal units regarded as bursting pace-maker cells; the total number of the septal cells secondarily (synaptically) involved into rhythmic activity. The effect of pentobarbital upon theta-rhythm results from elimination of these influences upon the septal cells.     

Electrical stimulation of hybridoma cells producing monoclonal antibody to cAMP

Electrical stimulation was applied to hybridoma cells in order to activate metabolic activities and increase the monoclonal antibody production. Hybridoma cells that produce monoclonal antibody to adenosine 3':5'-cyclic monophosphate were placed on a transparent glass electrode immersed in medium and subjected to electric pulses (pulse shape, alternating rectangular; field strength, 4 X 10(3) V X m-1; frequency, 5 kHz; pulse mode, 0.5 min application and 4.5 min pause). After 48 h of incubation, the concentration of lactic acid in the medium reached 8.4 mM, approx. 30% higher than that obtained without electric stimulation. Similarly, cell growth rate was promoted by the electric stimulation, reaching a maximum stimulation after 40 h. When the hybridoma was cultured for 48 h with electrical stimulation, the antibody concentration in the medium reached 22.3 microgram X ml-1, approx. 10% higher than the control, with a concomitant 16% increase in cell concentration. Longer periods of electric pulse application, however, caused an inhibitory effect on the hybridoma growth. The most probable cause of the inhibition are reactive oxygen species such as superoxide and hydrogen peroxide, which are inevitably generated by electrolysis. The presence of superoxide dismutase (EC 1.15.1.1) reduced the inhibitory effects. In conclusion, metabolic activities including monoclonal antibody production were activated by the electrical stimulation.     

Intracellular mechanism of action of sympathetic hepatic nerves on glucose and lactate balance in perfused rat liver

In rat liver perfused in situ stimulation of the nerve plexus around the hepatic artery and the portal vein caused an increase in glucose output and a shift from lactate uptake to output. The effects of nerve stimulation on some key enzymes, metabolites and effectors of carbohydrate metabolism were determined and compared to the actions of glucagon, which led to an increase not only of glucose output but also of lactate uptake. 1. Nerve stimulation caused an enhancement of the activity of glycogen phosphorylase a to 300% and a decrease of the activity of glycogen synthase I to 40%, while it left the activity of pyruvate kinase unaltered. Glucagon, similarly to nerve action, led to a strong increase of glycogen phosphorylase and to a decrease of glycogen synthase; yet in contrast to the nerve effect it lowered pyruvate kinase activity clearly. 2. Nerve stimulation increased the levels of glucose 6-phosphate and of fructose 6-phosphate to 200% and 170%, respectively; glucagon enhanced the levels to about 400% and 230%, respectively. The levels of ATP and ADP were not altered, those of AMP were increased slightly by nerve stimulation. 3. Nerve stimulation enhanced the levels of the effectors fructose 2,6-bisphosphate and cyclic AMP only slightly to 140% and 125%, respectively; glucagon lowered the level of fructose 2,6-bisphosphate to 15% and increased the level of cyclic AMP to 300%. 4. In calcium-free perfusions the metabolic responses to nerve stimulation showed normal kinetics, if calcium was re-added 3 min before, but delayed kinetics, if it was re-added 2 min after the onset of the stimulus. The delay may be due to the time required to refill intracellular calcium stores. The hemodynamic alterations dependent on extracellular calcium were normal in both cases. The activation of glycogen phosphorylase, the inhibition of glycogen synthase and the increase of glucose 6-phosphate can well explain the enhancement of glucose output following nerve stimulation. The unaltered activity of pyruvate kinase and the marginal increase of fructose 2,6-bisphosphate cannot be the cause of the nerve-stimulation-dependent shift from lactate uptake to output. The very slight increase of the level of cyclic AMP after nerve stimulation cannot elicit the observed activation of glycogen phosphorylase.(ABSTRACT TRUNCATED AT 400 WORDS)     

The generation of DPOAEs in the locust ear is contingent upon the sensory neurons

Tympanal organs of insects emit distortion-product otoacoustic emissions (DPOAEs) that are indicative of nonlinear ear mechanics. Our study sought (1) to define constraints of DPOAE generation in the ear of Locusta migratoria, and (2) to identify the sensory structures involved. We selectively destroyed the connection between the (peripheral) sensory ganglion and the tympanal attachment points of the “d-cell” dendrites; d-cells are most sensitive to sound frequencies above 12 kHz. This led to a decrease of DPOAEs that were evoked by f ₂ frequencies above 15 kHz (decrease of 15–40 dB; mean 28 dB; n = 12 organs). DPOAEs elicited by lower frequencies remained unchanged. Such frequency-specific changes following the exclusion of one scolopidial sub-population suggest that these auditory scolopidia are in fact the source of DPOAEs in insects. Electrical stimulation of the auditory nerve (with short current pulses of 4–10 μA or DC-currents of 0.5 μA) reversibly reduced DPOAEs by as much as 30 dB. We assume that retrograde electrical stimulation primarily affected the neuronal part of the scolopidia. Severing the auditory nerve from the central nervous system (CNS) did not alter the DPOAE amplitudes nor the effects of electrical stimulation.