The L3 neuron and an associated prothoracic network are involved in calling song recognition by female crickets

In young virginAcheta domesticus females, the spiking response of the prothoracic L3 auditory interneuron discriminates between calling songs (CSs) with phonotactically attractive and unattractive syllable periods (SPs), which parallels phonotactic discrimination. Presentation of a CS with an originally attractive SP, but with the intensity modulated so as to minimize L3's selective response, results in a CS with little phonotactic attractiveness. Conversely, a CS with an originally unattractive SP becomes much more attractive when the CS is intensity modulated in ways that duplicate L3's selective response. L3's discriminatory response to CS SP deteriorates with age, in parallel with decreased phonotactic selectiveness (females, older than 14 days, typically are unselective for CS SPs). SP-selective processing, which was not apparent in these old L3s, is immediately restored by removing the contralateral ear. SP-specific information is resident in a network of neurons within the prothoracic ganglion that results in the SP selective responses of the L3 neuron in young females. Changes in the SP-selective responses of the L3 neuron are highly correlated with corresponding changes in the female's phonotactically selective behavior.     

Nanoinjection of neurotransmitters into the prothoracic ganglion of female cricket Acheta domesticus changes phonotactic selectivity

The phonotactic response by female crickets is influenced by Juvenile Hormone III, which affects selectivity to the syllable period of the calling song. This pathway is influenced by an inhibitory input in the prothoracic ganglion, possibly chloride-mediated inhibition. In order to identify potential neurotransmitters involved in such pathway, we performed nanoinjection of five neurotransmitters into the prothoracic ganglion of virgin female Acheta domesticus. Phonotaxis for these females was evaluated before and after injections. All five neurotransmitters that were nanoinjected are known to bind to chloride channels. Nanoinjection of histamine significantly decreased phonotactic selectivity to the syllable period of the calling song while glycine, gamma aminobutyric acid, serotonin and saline controls did not. Octopamine significantly decreased phonotactic responses overall. The effect of histamine was tested further by nanoinjecting the antihistamine pyrilamine into the prothoracic ganglion of older unselected females, which resulted in increased phonotactic selectivity.     

Prolonged response to calling songs by the L3 auditory interneuron in female crickets (Acheta domesticus): intracellular evaluation

The L3 auditory interneuron in female Acheta domesticus, produces two different responses to the male calling song: an immediate response and a prolonged response. The prolonged response exhibited spiking activity and a correlated prolonged depolarization, both of which are clearly seen in intracellular recordings. The morphology revealed by intracellular staining was clearly the L3 neuron. The amplitude of the prolonged depolarization associated with the prolonged response increased with increases in sound intensity, resulting in increased spiking rates. Both depolarization and sound presentation increased the spiking rate and the slope of pre-potentials (thus leading to spiking threshold more quickly). Injecting hyperpolarizing current had the expected opposite effect. The effects of positive current injection and sound presentation were additive, resulting in spiking rates that were approximately double the rates in response to sound alone. Short postsynaptic potentials (PSPs), whose duration ranged from 15-60 ms, which may lead to action potentials were also observed in all recordings and summated with the prolonged depolarization, increasing the probability of spiking.     

Courtship song, male agonistic encounters, and female mate choice in the house cricket,Acheta domesticus (Orthoptera: Gryllidae)

Whether female crickets choose among males based on characteristics of the courtship song is uncertain, but in many species, males not producing courtship song do not mate. In the house cricket,Acheta domesticus, we examined whether a female chose or rejected a male based on his size, latency to chirp, latency to produce courtship song, or rate of the high-frequency pulse of courtship song (“court rate”). We confirmed that females mated only with males that produced courtship song, but we found no evidence that the other factors we measured affected a female’s decision to mate. In addition, we investigated whether the outcome of male agonistic encounters affected the subsequent production of courtship song. In one experiment, we observed courtship and mating behavior when a single female was placed with a pair of males following a 10-min interaction period between the two males. Winners of male agonistic encounters had higher mating success. However, winners and losers of agonistic encounters were not different in their likelihood or latency to produce courtship song or in the number of times they were disrupted by the other male in the pair. In a second experiment, we allowed two males to interact for a 10-min period, but following this interaction period, we placed a female with each male separately and observed courtship and mating behavior. The mating success of winners and losers was not different under these circumstances, and we found no differences between winners and losers in any subsequent courtship or mating behavior examined. We conclude that winning agonistic encounters influences a male’s mating success in ways other than his production of courtship song and this effect is lost when winning and losing males are separated and each is given an opportunity to mate.     

Processing of model calling songs by the prothoracic AN2 neurone and phonotaxis are significantly correlated in individual female Gryllus bimaculatus

Syllable period (SP) selective calling song processing has been demonstrated for the prothoracic, AN2 auditory neurone that correlates very well with SP‐selective phonotaxis by female cricket Gryllus bimaculatus De Geer. Both SP‐selective processing by the AN2 and the phonotactic behaviour of the female exhibit substantial plasticity. Thus, the question remains as to whether the selective responses of the AN2 neurone and the selective behaviour of the female match in an individual female. The present study is designed to answer that question. The SP‐selective phonotactic behaviour of individual females is evaluated, followed immediately by measuring the SP‐selective responses of the same female's AN2 neurone. Very significant correlations are found between the selective responses of the AN2 neurone and the same female's selective behaviour. In 208 possible comparisons (26 females, eight behavioural and neuronal tests each), 186 resulted in matches between behaviour and neuronal processing. Dividing the SP‐selective females into two groups (one group that responded phonotactically to the shortest SP tested and a second group that did not respond to this SP) resulted in significantly more selective responses to this shortest SP by the AN2 neurone in the females that responded phonotactically to the SP than for the females who did not respond to the shortest SP. The behavioural responses by these two groups to the other SPs tested are shown to be essentially identical.     

Morphological and electrophysiological characteristics of a dorsal unpaired median neuron of the cricket,Acheta domesticus

• 1.1. The dorsal unpaired median neuron (DUMDL) of the metathoracic dorsal longitudinal flight muscle was studied using cobalt and neutral red staining, various pharmacological agents and extracellular recording of electrical activity.
• 2.2. The soma of DUMDL is posterior in the group of dorsal unpaired median neurons of the metathoracic ganglion; DUMDL does not degenerate following histolysis of its muscle.
• 3.3. Slow, regular spontaneous activity of DUMDL is generated in the isolated metathoracic ganglion and is suppressed following stimulation of DUMDL and by discharges of the flight motor neurons. This spontaneous activity is increased by acetylcholine and eserine and blocked by anticholinesterase agents.
• 4.4. The neurite of DUMDL appears to have several impulse initiation zones characterized by a low safety factor.

     

The efferent innervation of the genital chamber by an identified serotonergic neuron in the female cricket Acheta domestica

Summary The serotonergic innervation of the genital chamber of the female cricket, Acheta domestica, has been investigated applying anti-serotonin (5-HT) immunocyto-chemistry at both light- and electron-microscopic levels as well as using conventional electron microscopy. Whole mount and pre-embedding chopper techniques of immuno-cytochemistry reveal a dense 5-HT-immunoreactive network of varicose fibers in the musculature of the genital chamber. All of these immunoreactive fibers originate from the efferent serotonergic neuron projecting through the nerve 8v to the genital chamber (Hustert and Topel 1986; Elekes et al. 1987). At the electron-microscopic level, 5-HT-immunoreactive nerve terminals, which contain small (50–60 nm) and large ( 100 nm) agranular vesicles as well as granular vesicles (100nm), contact the muscle fibers or the sarcoplasmic processes without establishing specialized neuromuscular connections. In addition to the 5-HT-immunoreactive axons, two types of immunonegative axons can also be found in the musculature. By use of conventional electron microscopy, three ultrastructurally distinct types of axon processes can be observed, one of which resembles 5-HT-immunoreactive axons. While the majority of the varicosities do not synapse on the muscle fibers, terminals containing small (50–60 nm) agranular vesicles occasionally form specialized neuromuscular contacts. It is suggested that the 5-HTergic innervation plays a non-synaptic modulatory role in the regulation circular musculature in the genital chamber of the cricket, while the musculature as a whole may be influenced by both synaptic and modulatory mechanisms.Fellow of the Alexander von Humboldt-Stiftung     

The effects of injecting Juvenile Hormone III into the prothoracic ganglion on phonotaxis by female crickets Gryllus bimaculatus

Nanoinjection of Juvenile Hormone III (JH III) into the prothoracic ganglion causes virgin female crickets Gryllus bimaculatus De Geer to become more phonotactically selective for the syllable periods (SPs) of model calling songs. Females responding to all, or almost all, of the SPs presented before JH III injection significantly narrow their responses to a range of SPs that is usually centered on the SPs included in the conspecific males' calling song. Control injections of acetone (i.e. the solvent for JH III) into the prothoracic ganglion do not significantly change the recipient females' phonotactic responses. Injection of JH III into the metathoracic ganglion also has no effect the females' phonotactic choices.     

Response properties of the prothoracic AN2 auditory interneurone to model calling songs in the cricket Gryllus bimaculatus

Sound processing properties for calling song (CS) models, as described for the prothoracic L3 auditory neurone in Acheta domesticus, are investigated for the homologous auditory neurone 2 (AN2) in female Gryllus bimaculatus De Geer. AN2 of G. bimaculatus responds selectively to the syllable period (SP) of models of a male CS. The selectiveness of this response parallels the selectivity of phonotaxis females perform in response to the same SPs. Both, the responses of AN2 and female behaviour show clear interindividual variability. The SP‐selective responses of AN2 result from an SP‐dependent reduction in the spiking to subsequent syllables of the model CSs, measured as the percentage decrement. This SP‐dependent response does not primarily result from inbuilt properties of the AN2 membrane. Rather, it is dependent on inhibitory input to the AN2. However, clear inhibitory postsynaptic potentials in dendritic recordings of the AN2 are not encountered. This immediate response of AN2 to CSs is followed by an increased rate of tonic firing between stimulus CSs, which is termed the prolonged response, and is dependent on the carrier frequencies that make up the male CSs. With stimulation on the contralateral side of the soma of AN2s, more than 50% of AN2s exhibit a prolonged response. However, with stimulation from the ipsilateral side of the soma, most AN2s exhibit a prolonged response. The prolonged response of AN2 at 5 kHz may be even more sensitive than the immediate response. Thus, the AN2 neurone could provide a basis for phonotaxis that is selective for both the SPs and the carrier frequencies of potentially attractive calling songs.     

Variance in female responses to the fine structure of male song in the field cricket, Gryllus integer

Although female mating preferences are a focus of current controversy,little detailed information exists on female preferences withinnatural populations. In the field cricket Gryllus integer, malecalls attract sexually receptive females, and females preferentiallymove toward male calls with longer calling bouts (periods ofcalling containing no pause greater than 0.10 s in real time).This study investigated female preferences for other variablesof the male song, including syllable period, chirp pause, andnumber of syllables per chirp. Male song was measured in thefield to determine mean values for each variable in nature.Female preferences were determined using a locomotor-compensatordevice, on which females ran in response to sequential playbacksof synthesized male song. Mean female preferences correspondedroughly to mean male song variables. Nonetheless, females variedgreatly in their responses to synthesized calls differing insyllable period, syllable number, and chirp pause. Moreover,individual females who were more selective for any one variablealso tended to be more selective for others. These results showthat females may differ from one another in their mating preferencesand degrees of selectivity, even within a single population.     

Divergence in the calling songs between sympatric and allopatric populations of the southern wood cricket Gryllus fultoni (Orthoptera: Gryllidae)

In the eastern United States the wood cricket Gryllus fultoni (Orthoptera: Gryllidae) occurs in sympatry with G. vernalis in an area between eastern Kansas and west of the Appalachian Mountains. Calling songs were recorded from 13 sympatric and allopatric localities. Both field and laboratory recordings showed that chirp rate (CR) and pulse rate (PR) overlapped extensively between allopatric populations of G. fultoni and sympatric populations of G. vernalis; by contrast, there was little or no overlap in these variables between sympatric populations of these two species. Divergence in PR and CR between the two species was thus greater in areas of sympatry than in areas of allopatry. Our field and laboratory studies of G. fultoni calling songs thus demonstrate the pattern expected of character displacement and support the genetic assumptions of this hypothesis. Other possible explanations for the sympatric divergence such as ecological character displacement and clinal variation are discussed.     

The effect of visual input on calling song attractiveness for female Acheta domesticus

ABSTRACT. Of twelve mature phonotactically-responsive female Acheta domesticus L., ten responded phonotactically to a wider range (30 or SO to 100 ms) of model calling song (CS) syllable periods (SP) on the Kramer treadmill in the dark than in a lighted visually structured arena (50–70 ms). When given a choice between the visually attractive target and the invisible loudspeaker, seven of the ten females that tracked a visually attractive target (black square) when presented alone in the light reduced the range of SPs they tracked phonotactically to 50–70 ms. Three of the ten females that were not strongly attracted to the visual target when presented alone, continued to respond to model calling songs with a wide range of SPs (30–100 ms) when given a choice between visual and acoustical targets under the same conditions. Two of the twelve females responded only to model calling songs with a 50–70 ms SP on the Kramer treadmill in the dark. These females did not change their choice for model calling song SPs when presented with the visually attractive target.     

Scototaxis and shape discrimination in the female cricket Acheta domesticus in an arena and on a compensatory treadmill

ABSTRACT. The scototactic responses of adult female Acheta domesticus L. were tested toward various shaped targets in an orientation arena and on a compensatory treadmill. In an arena, crickets oriented toward dark targets (positive scototaxis) if they had horizontal visual angles ≥30, and if the vertical dimensions of the target was ≤ its horizontal dimension. Unattractive targets did not result in negative scototaxis but caused crickets to orient randomly with no net directionality. When complex targets were composed of two or more simple rectangular targets which had been previously defined as attractive or unattractive, intermediate responses were obtained. Crickets oriented less toward complex targets than toward simple attractive targets, but oriented more than toward simple unattractive targets. The responses of a female cricket toward an attractive target can be modified by the presence of a chemical signal emitted by previously tested females. When the signal is present females are less likely to orient toward a target that would otherwise be very attractive. This effect was not as great when the females being tested were taken directly from densely populated colonies.
When running on a compensatory treadmill, female crickets exhibit scototactic tendencies similar to those displayed in the arena. When tested on the treadmill over long periods, the amount of time spent orienting toward an attractive target increased.     

Selective processing of calling songs by auditory interneurons in the female cricket, Gryllus pennsylvanicus: possible roles in behavior

Female crickets (Gryllus pennsylvanicus), caught in the field as nymphs, responded as adults in the laboratory with selective phonotaxis to model calling songs (CSs) that reproduced the dominant carrier frequencies and syllable periods (SPs) characteristic of the male's natural calling song. Extracellular recordings demonstrated two types of auditory interneurons in the female's cervical connectives that were very similar to the AN1 and AN2 neurons previously described in other gryllid species. The AN2 neuron responded to model CSs with a phasically encoded immediate response, and a more tonically encoded prolonged response. AN2's immediate response exhibited SP-dependent decreases (termed decrement) in its responses to sequential syllables of the CS that were greatest to CSs with the shortest SPs and diminished as SPs were lengthened, resulting in an SP-dependent habituation. Picrotoxin application transformed this SP-dependent habituation by AN2 to SP-selective responses in which the degree of decrement was greatest to SPs that were most phonotactically attractive. AN2's prolonged response was most sensitive to 5 kHz CSs and correlated with the carrier frequency tuning for the thresholds of phonotaxis by females. Thus, in females, AN2's immediate (in the presence of picrotoxin) and prolonged responses were selectively tuned to the SPs and carrier frequencies of the male's calls that were most attractive behaviorally. AN1's responses at threshold were also tuned to the dominant carrier frequencies of the male's CS.     

The energetic costs of fighting in the house cricket, Acheta domesticus L

The cost of performing an agonistic behavior, or tactic, willhave consequences for an individual's rate of cost accrual,the tactic's evolutionary stability if used as an assessmentsignal, and its pattern of use in the behavioral choreographyof a contest. Few studies have attempted to quantify the costsof fighting, particularly with regard to energy expenditure.Flow-through respirometry revealed that house cricket malescan expend energy at relatively high rates when fighting (morethan eight times resting levels) depending on the particulartactic performed. Acoustic signalling (stridulation) constitutedthe least costly of seven measured agonistic tactics, whilewrestling with an opponent, the most energetically costly tactic,consumed oxygen at a net rate more than 40 times that of stridulation.Low-cost tactics are used by opponents more frequently thanmore costly tactics, providing evidence for an escalating tacticalconvention based on energetic costs. The moderate energeticcost of some tactics suggests they may function as reliablesignals in the assessment of wrestling ability. Net energy costsper contest increased linearly with contest duration for bothcontest winners and losers, but winners tended to expend moreenergy per contest than losers. The likely fitness effects ofenergy expended while fighting are discussed. The results ofthis study indicate that energy expenditure is an importantcost shaping contest strategies in Acheta domesticus