J Comp Physiol A (2000) 186: 181±191 Ó Springer-Verlag 2000

ORIGINAL PAPER

H. RoÈmer á M. Krusch

A gain-control mechanism for processing of chorus sounds in the

afferent auditory pathway of the bushcricket Tettigonia viridissima
(Orthoptera; Tettigoniidae)

Accepted: 20 November 1999

Abstract The representation of alternative conspeci®c

acoustic signals in the responses of a pair of local
Introduction
interneurons of the bushcricket Tettigonia viridissima
Acoustic advertisement signals in orthopteran insects
was studied with variation in intensity and the direc-
are used to attract mates from a distance, or in com-
tion of sound signals. The results suggest that the
petitive interactions between males. However, signalling
auditory world of the bushcricket is rather sharply
often occurs in groups (choruses) of many individuals of
divided into two azimuthal hemispheres, with signals
the same and/or di€erent species. Whatever the proxi-
arriving from any direction within one hemisphere
mate and ultimate causes for chorusing may be (for re-
being predominantly represented in the discharge of
views see Alexander 1975; Otte 1977; Thornhill and
neurons of this side of the auditory pathway. In ad-
Alcock 1983; Green®eld 1994), the result of such ag-
dition, each pathway also selects for the most intense
gregations is a complex sonic and ultrasonic background
of several alternative sounds. A low-intensity signal at
over which individuals must then communicate. Thus,
45 dB sound pressure level is quite e€ective when
each species faces a trade-o€ between optimum broad-
presented alone, but completely suppressed when given
cast conditions and the problem of detecting and dis-
simultaneously with another signal at 60 dB sound
criminating individual conspeci®c signals against a high
pressure level. In a series of intracellular experiments
background noise.
the synaptic nature of the intensity-dependent sup-
Temporal segregation of calling activity (Gogala and
pression of competitive signals was investigated in a
Riede 1995) or the partitioning of calling frequencies
number of interneurons. The underlying synaptic
(Narins 1995) may represent solutions to reduce inter-
mechanism is based on a membrane hyperpolarisation
speci®c song interference, but insects may also sing in
with a time-constant in the order of 5±10 s. The sig-
dense populations of conspeci®c individuals, where more
ni®cance of this mechanism for hearing in choruses,
than one signaller is within hearing range of a potential
and for the evolution of acoustic signals and signalling
receiver. For example, the maximum hearing distance
behaviour is discussed.
for the calls of male bushcrickets Mygalopsis marki was
estimated at 25 m, a distance which, combined with a
Key words Bushcricket á Chorusing á Intensity á
map of male spacing in the habitat, suggests that more
Gain-control á Inhibition
than ten receivers may be within the active space of the
signal (RoÈmer and Bailey 1986). The temporal overlap
Abbreviations EPSP excitatory postsynaptic
of several songs arriving from di€erent directions may
potential á PSTH peristimulus time histogram á
result in a severe masking of the individual temporal
SPL sound pressure level
song pattern at the position of the receiver. Under such
conditions any estimation of the species-speci®c tempo-
ral properties, for example syllable or chirp duration or
intersyllable intervals, may be extremely hard to ac-
complish. The question, therefore, is how many of the
signals within hearing range of a receiver, if any, can be
H. RoÈmer (&) á M. Krusch evaluated in such choruses?
Karl-Franzens-University Graz, UniversitaÈtsplatz 2,
A-8010 Graz, Austria By mimicking part of such complex chorus situation
e-mail: roemer@kfunigraz.ac.at in the laboratory, Pollack (1988) discovered a neuronal
Tel.: +43-316-380-5596; Fax: +43-316-380-9875 gain-control mechanism in one of the pairs of omega
182
neurons (ON1) of the cricket that could selectively code
the more intense of two simultaneously presented sound
signals, analogous to the `cocktail party phenomenon'
familiar to humans (Cherry 1966). The intensity dif-
ferences between the competing sound signals used in
Pollack's study were 20 dB. The distribution of signal-
ling males in populations of crickets and bushcrickets
(Clark and Evans 1954; Campbell and Shipp 1979;
Thiele and Bailey 1980; Schatral et al. 1984; Forrest
and Green 1991; Arak and Eiriksson 1992) would
suggest, however, that the acoustic situation for either
male or female receivers in such populations may be far
more complex in both the number of possible sound
sources within hearing range and the intensity di€er-
ences that need to be discriminated. The aim of the
present paper is therefore twofold. First, we con®rm
and detail a synaptic mechanism similar to the one
described in the cricket and in the bushcricket Tetti-
gonia viridissima (RoÈmer 1993). Second, we employ
®eld data from populations of the same insect to apply
a more natural signal situation in neurophysiological
tests for the functional signi®cance of such a neuronal
mechanism in nature. The results of both ®eld and
laboratory studies on the representation of competitive
acoustic signals provide further evidence for the exis-
tence of a mechanism for the suppression of responses
to the weaker of two or more sounds. The reported
proximate mechanisms of sensory processing in the
a€erent auditory pathway have important implications
for the evolution of acoustic communication in this
group of insects.
Material and methods
Field studies
Measurements of acoustic conditions for male or female receivers
were performed in three populations of the bushcricket T. vi-
ridissima, between July and September 1990 and 1993. The popu-
lations were selected because the type and density of the vegetation
was rather similar. The study sites were either in the vicinity of the
Ruhr-University Bochum, or Karl-Franzens-University of Graz,
each with an area of about 1 ha. The vegetation consisted of a
fairly homogeneous layer of grass (<0.8 m), interspersed with
patches of taller vegetation with a maximum height of 2.5 m.
We censused the three sites during periods of continuous sun-
shine, between 1600 hours and 1830 hours, which usually was
about 2 h after the initiation of singing. At this time, most of the
males had established their preferred singing sites. Three people
located a singing male from a distance, as well as up to four of its
nearest neighbours without disturbing their singing activity. After
the positions of the focal and surrounding males had been identi-
®ed, one person silenced the focal male and measured the sound
pressure level (SPL) of the songs of each neighbouring male at the
focal male's position, using a portable sound level meter (Bruel &
Kjaer Model 2009, 1/2¢¢ microphone, type 4133; RMS fast reading).
SPL values are expressed in decibels re. 20 lPa. For each mea-
surement the microphone pointed in the direction of the respective
singing neighbour. In those cases were the singing activity of one
neighbour interfered with the SPL measurement of another, we
either silenced one male or waited for intervals within the long-
lasting trills of sucient length to obtain reliable SPL readings for
the desired individual.
Laboratory studies
Neurophysiology
Males or females were anaesthetised with CO2, and after removal
of the wings, mid- and hindlegs, were mounted ventral side up on
a small metal holder using dental wax. The prothoracic legs
bearing the hearing organs were ®xed in a position 90° to the
longitudinal body axis. The prothoracic ganglion was exposed, the
gut underneath removed and replaced by a small piece of cotton
to prevent movement of the ganglion during long-term recordings
from the bilateral pair of omega cells. These cells are local neu-
rons in the prothoracic ganglion, receiving direct excitatory input
from most receptors in the hearing organ (RoÈmer et al. 1988; see
Fig. 3A). The extracellular spike activity of the pair of cells was
recorded with glass-insulated tungsten electrodes (resistance
1±3 MW) inserted into the anterior part of the ganglion. By po-
sitioning the tip of the electrode close to the crossing segments of
both cells and slightly o€ the ganglionic midline, we obtained
spike recordings of both cells with reliably di€erent amplitudes
(Fig. 3A; for similar results with the pair of ON1 neurons in
crickets see Wiese and Eilts 1985). The crossing segments of the
two cells form the so-called omega tract, with no axon or dendrite
of other auditory cells entering (Wohlers and Huber 1985). As a
rule the soma-contralateral cell had the larger action potential
amplitude, which was controlled by changing the position of the
speaker from one side to the other. It is still unknown why the
contralateral omega cell always exhibits the larger action poten-
tials in extracellular recordings. Despite intensive extra- and in-
tracellular recordings made in the prothoracic ganglion of
bushcrickets in our laboratory over many years, we know of no
other pair of cells that allows simultaneous spike recordings with
an extracellular electrode at this position. Thus, although our
recordings were only extracellular and no systematic morpholog-
ical identi®cation of the recorded cells were made, we are con®-
dent that all extracellular recordings were made from the pair of
omega neurons.
Once the di€erence in spike amplitude was large enough for
separate analysis of the two cells, the opening in the cuticle was
sealed with Vaseline to prevent desiccation of the ganglion. The
preparation was placed in the centre of an anechoic chamber such
that echoes at the position of the preparation were more than
32 dB less intense than the signal. Thus, with a standard SPL of
60 dB used in the experiments, echoes would be just at the
threshold level of the omega neurons.
In a second series of experiments the omega cell and a number
of other interneurons responding to sound were also recorded in-
tracellularly to study the synaptic events underlying the mecha-
nisms of selective responses to particular sound signals.
Morphological identi®cation of the cells was by subsequent stain-
ing with lucifer yellow using conventional techniques (Steward
1978). Sixteen omega cells and 18 other interneurons were studied
intracellularly.
Sound stimulation
The three song models (Fig. 1C; patterns 1±3) were constructed
from recordings of the calling song of an isolated T. viridissima
male, using a 1/4¢¢ microphone (Bruel & Kaer, type 4136), pre-
ampli®ed through a sound level meter (Bruel & Kjaer, type 2209).
The signal was digitised from the tape recorder (Racal store 4D)
on a custom-made AD/DA converter (12 bit, sampling rate of
125 kHz). The song consists of a train of double syllables, re-
peated monotonously over long periods of time (Fig. 1A). When
analysed close to the male, the power spectrum of the song re-
veals a broad energy distribution between 8 kHz and 50 kHz
(Rheinlaender and RoÈmer 1980; Keuper et al. 1988; Arak and
Eiriksson 1992). However, at the distances corresponding to
normal spacing of the males, excess attenuation reduces the rel-
ative amplitudes of the higher ultrasonic components (Keuper
and KuÈhne 1983; RoÈmer and Lewald 1992). We therefore used a

Fig. 1A±C Oscillogram of the calling song of a male T. viridissima.
Scale bar in A and C 100 ms. Song models (patterns 1±3) used in
playback experiments (C) were constructed from digitised songs as
shown in A with di€erent syllable intervals. The low-amplitude
hemisyllables as a result of opening the wings (small amplitude pulses
in A) were omitted in the model songs. B Power spectrum of the song
models. For further explanation see text
song model with a spectral composition re¯ecting intermediate
distances (Fig. 1B).
One of the double syllables in the song was digitally cut and
repeatedly copied into a series of ten double syllables. By using
di€erent intersyllable intervals, we constructed three di€erent pat-
terns of song which were identical in energy, spectral composition
and total length (675 ms), but di€ered in their temporal pattern
(Fig. 1C). These di€erent temporal patterns facilitate the identi®-
cation of response patterns in complex stimulus con®gurations
using correlation analysis. In a series of control experiments we
con®rmed that the omega cell shows no preference for any of the
three temporal patterns, given the SPL is the same (for similar
results with the omega neuron in crickets see Schildberger 1984).
Pattern 1 was broadcast at a rate of 1/s through the D/A converter
of the PC (12-bit resolution; 125-kHz sampling rate), while patterns
2 and 3 were recorded on tape (Racal Store 4D) at periods of
1010 ms and 990 ms, respectively. Thus, when pattern 1 was
broadcast together with pattern 2 or 3 this resulted in a quasi-free,
¯oating phase relationship between stimuli, when a stimulus con-
®guration was repeated 40 times for later correlation analysis of the
action potential discharges (see below).
Each song pattern was broadcast through separate wide-band
ampli®ers (Akustischer Stimulator II, Burchard) and speakers
(TW8 special), mounted on holders which could be moved from
outside the anechoic chamber in calibrated steps of 7.5°, 15° or 30°
in the anterior semicircle surrounding the preparation, at a distance
of 45 cm. In a three-stimulus paradigm, a third speaker was either
placed at a given angle in the anterior hemicircle, or at a height of
40 cm above the preparation, in order to present a stimulus sym-
metrical for both sides. The SPL of each signal, controlled by
separate attenuators and measured at the position of the prepara-
tion, is given as root-mean-square (RMS, fast reading) SPLs (re:
20 lPa).
In 10 of 26 recordings of the pair of omega neurons the sym-
metry of tuning and directionality was investigated before each
two-stimulus experiment. Sound pulses with di€erent pure tone
183
carriers (2±50 kHz, standard duration 50 ms; repetition rate 1 s)1)
were generated to study the tuning of each cell with ipsilateral
stimulation. Stimuli were ampli®ed and attenuated in steps of
10 dB or 1 dB (Akustischer Stimulator II, Burchard). The thresh-
old was determined according to a procedure described in detail by
Rheinlaender and RoÈmer (1980). The symmetry of the directional
responses was studied in a single stimulus paradigm, in which a
single temporal pattern (usually pattern 1) was presented from
di€erent speaker positions in the frontal hemicircle. Increments of
30° (in the frontal zone 15° and 7.5°) were used.
Data analysis
Action potentials were recorded on a tape recorder (TEAC A-
3440) in parallel with the trigger(s) for the stimuli. The discharges
of both cells were detected by a window discriminator and ana-
lysed as peristimulus time histograms (PSTHs; bin width 2 ms). In
controls we estimated the error in discriminating action potentials
of both cells due to superposition of two spikes to be in the order
of 1% of the total spike activity. For the construction of histo-
grams each stimulus con®guration was presented for 50 s, where
the analysis of spike trains was restricted to the last 40 stimulus
presentations within a series in order to prevent any e€ects re-
sulting from ongoing activity after periods of silence. In prelimi-
nary experiments 40 stimulus representations turned out to be
sucient for the construction of PSTHs and reliable correlation
analysis. The similarity between the temporal pattern of a given
stimulus and the recorded spike train of the pair of omega neurons
was quanti®ed using correlation analysis (for details of the method
see Pollack 1988). The value between 0 and 1 gives a quantitative
measure of the degree to which a given stimulus pattern is rep-
resented in the spike pattern of a neuron. For example, if the
neuron is just responding to random noise the correlation coe-
cient with one of the stimulus patterns usually was between 0.08
and 0.12.
Results
Field measurements
The results of the ®eld measurements are summarised in
Fig. 2, in which data for the three populations are
pooled. SPLs of neighbouring males as received by a
focal male are shown in Fig. 2A; they reveal a broad
distribution between 50 dB and 84 dB SPL, with a mean
of 62 dB SPL. The intensity di€erence between the sig-
nals of neighbouring males appears to be important for
the receiver's ability to distinguish between these. Al-
though di€erences of up to 20 dB were recorded, they
are usually rather small (mean 4 dB; Fig. 2B).
Laboratory experiments
The degree to which di€erent spatially separated sound
stimuli are represented in a bilaterally organised sensory
system depends very much on the morphological and
neuronal symmetry of such a system. Peripheral or
central nervous asymmetries may result in a false rep-
resentation of such stimuli and thus misleading phono-
tactic behaviour or discrimination between signals. One
measure of auditory symmetry comes from the sensory
thresholds of bilaterally homologous elements within the
central nervous system. We therefore compared auditory
184

stimulus was broadcast at a medium intensity of 60 dB

SPL from the ipsilateral side of the left omega neuron
and then at various angles in the frontal hemicircle of
the preparation up to the ipsilateral side of the right
omega neuron. Analysis of the simultaneously recorded
responses of both cells indicates a strong preference in
response magnitude of the neuron stimulated from its

Fig. 2 A Sound pressure levels (SPLs) of neighbouring males as

perceived by a focal male, pooled from three populations (n ˆ 132).
They reveal a broad distribution between 50 dB and 84 dB SPL, with
a median value of 62 dB SPL. B Intensity di€erence between the
signals of neighbouring males at the position of a focal male (mean
4 dB). C Masking of the temporal pattern of neighbouring males at
the position of a female. Upper line: three nearest neighbours singing;
lower line: two of the three neighbours were silenced

threshold curves in ten pairs of omega neurons.

Thresholds of right and left omega cells are usually very
similar; in eight of the ten preparations thresholds did
not deviate by more than 2 dB at any frequency. The
maximal di€erence in the remaining two preparations
was 12 dB.
The symmetry of directionality of both cells was
studied in a single-stimulus paradigm. A conspeci®c

c
Fig. 3 A Schematic diagram demonstrating the method of recording
and discriminating the spike activity of the pair of omega neurons.
Action potentials of di€erent amplitude were obtained from the
crossing segments of the cells within the so-called omega tract
(Wohlers and Huber 1985) using an extracellular tungsten electrode of
low resistance (1±3 MW). The soma-contralateral cell was generally
the one with the larger spike amplitudes in the extracellular
recordings. The camera lucida reconstruction of the cell was obtained
from an intracellular staining with lucifer yellow. Scale bar 100 lm.
B Symmetry of directional responses of a pair of omega cells, as
revealed by a conspeci®c stimulus presented at 60 dB SPL from
various angles of sound incidence in the frontal hemisphere of the
preparation. In this and the following ®gures a schematic diagram of
the stimulus con®guration is given on top. Note the symmetrical,
mirror-image response behaviour of both cells along the longitudinal
body axis, and the steep change in response magnitude within a
narrow frontal zone (stimulus angle of ‹15°). Signi®cantly di€erent
responses occur with stimulus angles as small as 7.5° on either side of
the longitudinal body axis. Seven other pairs of omega cells were
tested in the same way and revealed similar results regarding the steep
slope and switch in activity in the frontal zone of the insect. However,
the symmetry axis coincided only in three out of eight preparations
with frontal stimulation, whereas in the remaining pairs this axis could
be shifted by 10° to either side of the body. C The peristimulus time
histograms (PSTHs) of the responses indicate that the temporal
features of the stimulus are faithfully encoded in the omega neurons
stimulated ipsilaterally

respective ipsilateral side, with a rather symmetrical,
mirror-image response behaviour of both cells along the
longitudinal body axis (Fig. 3B). More importantly, the
steepest slope of the curves occurs with a frontal stim-
ulus angle of ‹15±30° and signi®cantly di€erent re-
sponses were recorded with stimulus angles as small as
7.5° on both sides of the longitudinal body axis. The
PSTHs of the responses in Fig. 3C further indicate
that the temporal features of the stimulus are faithfully
Fig. 4A, B Representation of two sound signals (patterns 1 and 2) in
the spike discharge of the pair of omega neurons recorded
simultaneously (n ˆ 8). The SPL of each signal was 60 dB, the
position of the two speakers was varied from 90° right and left to 7.5°
in front of the preparation. In this and the following ®gures the
stimulus patterns are presented simultaneously, but due to their
slightly di€erent intervals are shifting in time relative to the others.
A The correlation coecient gives a quantitative measure of the
degree of similarity between the spike discharge and the temporal
pattern of the signals. Note the consistent representation of the
ipsilateral signal in the spike trains, even with an angular separation of
only 15° in the frontal zone. This is also evident in the PSTH of the
right and left omega neuron discharges with an angle of 7.5°.
B PSTHs of the responses of both cells, triggered with either signal
pattern 1 or 2. Note the reliable temporal coding of the ipsilateral
signal with a stimulus angle of 7.5°. The patterned response to the
respective contralateral signals is due to their inhibitory action on the
spike discharge
185
encoded in the omega neurons stimulated ipsilaterally,
irrespective of whether the stimulus angle was 90° or 30°.
In a two-stimulus paradigm we tested the ®delity of
temporal pattern encoding in the pair of omega neurons,
and how this ®delity is a€ected by changes in the spatial
con®guration of the stimuli (Fig. 4). When two stimulus
patterns are presented simultaneously, one on each side
at right angles to the body axis, the ipsilateral stimulus is
faithfully represented in the spike pattern of the omega
neuron on that side. This is also demonstrated qualita-
tively in the PSTHs for other stimulus angles of 30°, 15°
and 7.5° in Fig. 4B, in which pattern 1 is encoded in the
left omega neuron, and pattern 2 in the right omega
neuron even when the two sound sources are separated
by stimulus angles as small as 7.5° o€ the longitudinal
body axis. The quantitative measure of the correlation
coecient reveals almost no change in the quality of this
representation up to an angle of 15°, although even with
a total angular separation of only 15° in the frontal zone
there is still a signi®cantly better representation of ipsi-
lateral stimuli in the discharge of the omega neurons
(Fig. 4A). This is also evident in the PSTH of the right
and left omega neuron discharges with the angle of 7.5°.
A further complication of the receiver situation was
tested using an additional signal to the stimulus para-
digm, and quantifying the representation of all three
sound signals in the spike discharge of both omega
neurons (Fig. 5). In these experiments, the SPL of each
signal was again kept at 60 dB. Two speakers, broad-
casting patterns 1 and 3, were positioned 90° ipsilateral,
while pattern 2 was presented from varying angles of
sound incidence in front of the preparation. The quan-
titative measure of the correlation coecient for the
quality of signal representation reveals a mirror-image
when the pattern 2 signal is switched from left to right.
For example, there is an almost exclusive representation
of pattern 1 in the left omega neuron, when both other
patterns are broadcast from the right side (Fig. 5A).
When pattern 2 was broadcast as well from the left side,
the representation of both patterns is almost equal
(correlation coecient of 0.5). By contrast, the repre-
sentation in the opposite omega neuron mirrors that of
its counterpart (Fig. 5B): almost exclusive representa-
tion of pattern 3 when both pattern 1 and 2 were given
on the left side of the animal, and equal representation
for both when they are on the right side. Signi®cant
di€erences in the representation of two ipsilateral signals
do occur when one is broadcast from within an 15° angle
of sound incidence in front of the preparation, and the
other one at 90° (arrows in Fig. 5).
The ®ndings summarised in Figs. 4 and 5 suggest that
the auditory world of T. viridissima is rather sharply
divided into two azimuthal hemispheres, with signals
arriving from any direction within one hemisphere being
predominantly represented in the corresponding side of
the auditory pathway. A direction-selective representa-
tion such as this suggests a signi®cant contribution of
side-dependent binaural inhibitory interactions, which
are known to play a major role in the directionality of
186

Fig. 5 Representation of three

sound signals (patterns 1, 2 and
3) in the spike discharge of a
pair of omega neurons (left A;
right B) recorded simulta-
neously (n ˆ 7). The SPL of
each signal was 60 dB, the
position of two speakers (pat-
terns 1 and 3) was kept constant
at 90° ipsilateral, while pattern
2 was presented from varying
angles of sound incidence in
front of the preparation. Note
the identical quantitative mea-
sure of signal representation
when two signals of the same
SPL are presented on the ipsi-
lateral side. For further expla-
nation see Fig. 5 and text

auditory neurons in grasshoppers and crickets (review

by Pollack 1998). We therefore investigated the
responses of the omega neuron in a two-stimulus para-
digm, when the auditory input to the contralateral side
was eliminated by cutting the leg nerve carrying the
a€erent axons of auditory receptors. The results obtained
in eight preparations indicate that even in the unilateral
system the sound pattern broadcast ipsilaterally is sig-
ni®cantly better represented compared to a simultaneous
pattern on the contralateral side (Fig. 6). This is true for
all angles of sound incidence tested up to 7.5°.
The situation for a receiver in a population of many
calling males may, however, be more complicated than
just separating two sounds from opposite directions.
Two or more sound signals may arrive from di€erent
distances and thus with di€erent SPL, but from direc-
tions within one hemisphere of the animal. We therefore
conducted a series of experiments where two song pat-
terns of di€erent SPL were broadcast from the same
direction, and the representation of these patterns within
the spike trains of the ipsilateral omega neuron was
again investigated using correlation analysis. Figure 7A
shows the result of one typical experiment with one song Fig. 6 Representation of two sound signals (patterns 1 and 2) in the
pattern broadcast at a constant intensity of 60 dB SPL, spike discharge of one omega neuron, after the contralateral auditory
system was dea€erented by cutting the contralateral leg nerve (n ˆ 8).
while the playback SPL of the other pattern was varied SPL of each signal was 60 dB, the position of the two speakers was
from 30 dB to 70 dB. Both patterns were broadcast varied from 90° right and left to 7.5° in front of the preparation. For
from the same 90° direction, so no direction-selective further explanation see Fig. 5
component can be involved in the responses. This would
mimic a situation in nature where one male is closer to
 187

Fig. 7A, B Representation of

two sound signals (patterns 1
and 2) in the spike discharge of
one omega neuron, when both
signals are presented from the
same, ipsilateral side. The SPL
of pattern 1 was maintained at
60 dB, while that of pattern 2
was varied from 30 dB up to
70 dB. A Bilateral intact sys-
tem; B unilateral system after
cutting the contralateral leg
nerve (n ˆ 8). The correlation
coecient of the spike dis-
charge in response to pattern
2 at 45 dB SPL presented alone
is given for comparison
(arrows). Note the almost com-
plete suppression of the less
intense signal between 30 dB
and 45 dB SPL when there is a
competitive signal at 60 dB SPL

the receiver than another, or is calling from an elevated
perch and thus more e€ectively, but both from the same
direction. When the low-intensity song was broadcast
with SPLs between 30 dB and 45 dB simultaneously
with another song at 60 dB SPL, the representation of
the former was completely suppressed, although the low-
intensity song was quite e€ective at 45 dB SPL (or more)
when presented alone. This is evident in the di€erence of
the correlation coecient for the representation of pat-
tern 2 at 45 dB SPL when broadcast alone, compared to
simultaneous broadcast with pattern 1 (arrow in
Fig. 7A). At still higher intensities the correlation coef-
®cient for pattern 2 increases continuously, and with the
same SPL of 60 dB as pattern 1, exhibits the same
quantitative value for its representation within the dis-
charge of the ipsilateral omega neuron. Remarkably,
below 60 dB SPL intensity di€erences as small as 2±5 dB
resulted in a signi®cantly better representation of the
more intense signal compared to the less intense signal.
Above 60 dB SPL, however, the di€erences in the ®-
delity of representation were less pronounced even with
intensity di€erences of 10 dB.
Since the two song patterns were broadcast from the
same, ipsilateral direction it is very unlikely that lateral
inhibitory interactions could be responsible for this kind
of selectivity. Indeed, control experiments with the uni-
lateral system (contralateral leg nerve with tympanal
a€erents cut) revealed the same degree of selectivity for
intensity di€erences in the order of 2±5 dB (Fig. 7B). In
these unilateral experiments the dea€erented omega cells
exhibited little spontaneous activity (less than 2 APs/s
on average), and therefore its in¯uence on the activity of
the cell of the intact side through reciprocal inhibition
should be of no relevance.
In a series of intracellular experiments we investigated
the synaptic nature of the intensity-dependent suppres-
sion of competitive signals in the omega neuron, and
several other interneurons of the prothoracic ganglion.
Figure 8 shows two representative examples of intra-
cellularly recorded responses from an omega neuron
stimulated with long-lasting signals (pattern 1). The
synaptic events resulting in suprathreshold spike activity
consist of two components; excitatory postsynaptic po-
tential (EPSP) activity induced by each train of syllables,
and an additional inhibitory component most prominent
at higher intensities as a slowly developing increase in
the membrane potential of the cell. Starting from a
resting membrane potential of )52 mV, the cell is
hyperpolarized by 9 mV to )61 mV after about 5±10 s
of stimulation. The time-course of the inhibition is
extremely slow and it may take more than 10 s of intense
stimulation to reach the maximum amplitude of the
inhibition. Similarly, at the end of stimulation, it may
require 15 s or more to re-establish the resting mem-
brane potential and the level to elicit spontaneous action
potentials, depending on stimulus intensity and time of
stimulation (see Fig. 8; 70 dB SPL). Maximal values of
Fig. 8 Intracellularly recorded activity of an omega neuron in
response to a model of a conspeci®c song (pattern 1) at 40 dB and
70 dB SPL. Spikes are truncated to emphasise the biphasic synaptic
events underlying the response. Note the development of the slow
membrane hyperpolarization at the onset of the signal, and its decay
when the signal is switched o€. Note also the low membrane noise at
the end of the stimulation period (arrow in the upper recording) and
its increase with the return to membrane resting potential (compare
with Fig. 9C)
188
this hyperpolarizing potential varied from 5 mV to
12 mV between omega cell recordings. The intensity-
dependent expression of an inhibitory potential under-
lying the additional excitation of the neuron would
explain why, in a competitive situation of two or more
sound signals, the most intense signal is favoured in the
response of the neuron (see extracellular results in
Fig. 7). In a situation of strong membrane hyperpolar-
ization it is only those EPSPs which are elicited by the
most intense sound signals which may reach spiking
threshold of the neuron, whereas those induced by the
less intense signal(s) remain subthreshold.
In the course of our intracellular experiments we also
recorded activity from a number of interneurons as-
cending to the brain, which were tested in a similar way
with simultaneously presented sound signals of di€erent
SPL. For the purpose of this study they were roughly
categorised into tonic cells which represented the long-
lasting conspeci®c song pattern in their spike response,
and phasic neurons which habituated rapidly after few
double syllables of the song. As a rule, all tonically re-
sponding cells exhibited the same kind of biphasic re-
sponse as the omega cell, i.e. they responded with EPSPs
(and spikes), but also with a long-lasting underlying
hyperpolarizing potential. The degree of hyperpolariza-
tion varied between interneurons, but even a moderate
hyperpolarization showed the e€ect of suppression of an
alternative, low-intensity sound signal in the supra-
threshold spike response. In Fig. 9A the responses of
such a neuron demonstrate the biphasic character, with
Fig. 9 A Intracellularly recorded activity of an interneuron (an2 after
RoÈmer et al. 1988) within the prothoracic ganglion with an axon
ascending to the brain (OSG), in response to stimulation with a
conspeci®c song (pattern 1) at 55 dB SPL, and pure tone bursts at a
rate of 5 s)1 (20 kHz; 50 dB SPL) through a second, ipsilaterally
placed speaker. Note the reduction in excitatory postsynaptic
potential (EPSP) amplitude at the end of the stimulation series with
the more intense stimulus (B higher temporal resolution). C
Amplitude of spontaneous PSPs at the membrane resting potential
(upper line) and after prolonged activation with a conspeci®c signal
(lower line)
EPSP and tonic spike discharge, as well as a moderate
increase in membrane potential of 4±5 mV when the
preparation was stimulated for 14 s with a conspeci®c
song (pattern 1) at 55 dB SPL. Through a second
speaker the preparation was continuously stimulated
with pure-tone, 20-kHz sound bursts at a rate of 5 s)1, at
50 dB SPL, which resulted in EPSP and spike activity
before and after the conspeci®c song was switched on.
When the two sound signals coincide in time, however,
the EPSPs evoked by the low-intensity signal are both
reduced in amplitude and prevented from reaching the
spiking threshold due to hyperpolarization of the
membrane. This can be seen most clearly at the end of
the stimulation series with the high-intensity stimulus
(see inset in Fig. 9B), where the amplitudes of EPSPs
evoked by the tone bursts are small and remain sub-
threshold under the activated hyperpolarized situation,
but increase continuously until they reach normal spik-
ing level.
Further indications for the inhibitory action of the
slow membrane hyperpolarization comes from obser-
vations of spontaneous PSPs at the membrane resting
potential before stimulation, and when the cell is hy-
perpolarized due to prolonged activation with a con-
speci®c signal (Fig. 9C). In the resting state the neuron
shows membrane ¯uctuations as a result of spontaneous
PSPs with amplitudes up to 2±3 mV (Fig. 9C, upper
line), whereas immediately at the end of a 5-s period of
stimulation with conspeci®c song such PSP amplitudes
are completely absent (lower line). Finally, we injected
0.8-nA negative current pulses (duration 2 ms, rate
100 Hz) through the intracellular recording electrode
and a balanced bridge circuit into the neuron. We thus
observed an increase in membrane conductance at the
end of a long period of stimulation with conspeci®c
song, in comparison with the pre-stimulus situation.
Such increase in membrane conductance is a further
indication of the inhibitory nature of the membrane
hyperpolarization.
Discussion
Field measurements in populations of the bushcricket
T. viridissima revealed that the problem for a receiving
insect may be quite complex, requiring them to dis-
criminate the individual calls of up to four nearby males
and more than ten others within hearing range, some of
which are quite similar in amplitude. With this back-
ground information from ®eld populations, we studied
the response of auditory interneurons to playbacks mi-
micking the complex receiving situation in laboratory
experiments, extending those on cricket hearing per-
formed by Pollack (1988). The technique of recording
the discharges of both omega cells simultaneously
(Wiese and Eilts 1985) enabled us to track the repre-
sentation of alternative signals on both sides of the au-
ditory pathway. One result is that the auditory world of
the bushcricket is sharply divided into two hemispheres,

with a pronounced separation along the longitudinal
body axis. Even with an angular separation of only 7.5°
o€ the midline axis, two sound signals are signi®cantly
better represented in the spike discharge of the omega
neuron on that side. This separation is much stronger
than that of the cricket (Pollack 1988), probably as a
result of a steeper peripheral directionality of the ear. A
change in the angle of sound incidence of 30° in the
frontal zone (15° left to 15° right) results in a relative
change of 2.5±3 dB in the SPL of a conspeci®c cricket
signal (Rheinlaender 1984; Michelsen et al. 1994),
compared to 4.5 dB in the bushcricket T. viridissima
(Krusch 1991) and 5 dB for Leptophyes punctatissima
(Rheinlaender et al. 1986). Except for the study by Mi-
chelsen et al. (1994) the peripheral directionality was
studied neurophysiologically using unilaterally dea€e-
rented preparations. This higher peripheral directional-
ity, combined with the process of lateral inhibition, may
be sucient to explain the highly developed side-de-
pendence of two sound signals within right and left au-
ditory pathways.
Although each auditory pathway is selectively lis-
tening to, and encodes the temporal pattern of
predominantly ipsilateral sounds, each pathway also
selects for the most intense of several alternative sounds.
The underlying synaptic mechanism, ®rst described in
the cricket omega neuron by Pollack (1988), is based on
the fact that the response of the omega neuron to a
calling song comprises two components. In addition to
exciting the neuron, the signal also causes an inhibition
which can be seen as a hyperpolarisation with a slow
build-up and decay time. The time-constant in the order
of 5 s is most probably the major reason for the late
discovery of such a mechanism, despite extensive studies
on the auditory system of ensiferan insects. T. viridissima
and a number of other bushcrickets and crickets produce
highly repetitive songs which may last for very long
periods of time, many seconds, minutes or even hours.
In most previous physiological experiments, however,
our own included, although calling songs with ``natural''
syllable structure and frequency spectrum were used,
these generally lasted only some 10±100 ms, which
would preclude the identi®cation of the membrane hy-
perpolarization.
The source of the inhibition is certainly ipsilateral,
since it was also found in monaural preparations in
which the contralateral leg nerve with the axons of re-
ceptor ®bres was cut (Fig. 7B). A molecular mechanism
for the inhibition has been identi®ed in the omega neu-
ron of the cricket Acheta domesticus as an increased
intracellular free calcium concentration in response to
simulated cricket calling songs (Sobel and Tank 1994).
Evidence was obtained that the action of calcium is
through a calcium-dependent potassium conductance.
The time-constant of the increase in intracellular free
calcium correlates with the development of the slow
hyperpolarization in the same neuron. The gross anat-
omy of the omega neuron in T. viridissima and a number
of other bushcrickets, the existence of a separate omega
189
tract for the midline-crossing segment of the cell, and the
pattern of connectivity with primary a€erents (RoÈmer
et al. 1988) strongly suggest that it is homologous with
the omega cell in the prothoracic ganglion of crickets
(Wohlers and Huber 1978; Selverston et al. 1985). This
would imply that the gain-control mechanism in the
form of the slow hyperpolarization in this neuron is
common to many, if not all, ensiferans with implications
for both intra- and interspeci®c communication, which
have been discussed in detail by RoÈmer (1993).
Based on the properties of the gain-control mecha-
nism in sensory interneurons we can make two
straightforward predictions for the evolution of acoustic
signals. First, we would expect that louder males will
attract a greater number of females simply because (1)
the active range of their signal is greater, and (2) their
signal inhibits the representation of competitive signals
if the di€erence in loudness is 2±5 dB. Thus, selection
would act on males to produce or broadcast more and
more intense signals until the bene®t of increased loud-
ness is balanced by the costs of producing the signals,
whatever these costs are. Di€erential attraction of fe-
males to louder calling songs is known for many insects,
and also for frogs and toads. For example, one of the
few ®eld studies showed that the number of ¯ying male
and female mole crickets (Scapteriscus acleatus) at-
tracted to calling males in outdoor arenas was in¯uenced
by the SPL of the song relative to that of others calling
in the arena. Males calling more than 2 dB below the
loudest male attracted fewer individuals per male than
the overall average (Forrest and Green 1991). Virtually
all studies performed on acoustic insects support the
idea that there is strong selection on the signals or sig-
nalling behaviour to maximise broadcast range. Fruit-
¯ies, mole crickets, crickets and bushcrickets all
preferentially approach the louder of two conspeci®c
signals (review by Ryan and Keddy-Hector 1992). Se-
lection has also favoured amplifying mechanisms for the
signal and behaviour(s) which maximise calling range
(see Forrest 1994 and RoÈmer 1998 for reviews). Since the
gain-control mechanism enhances the di€erential repre-
sentation of competitive signals as a result of di€erences
in their SPL as small as 2 dB, it may serve as an addi-
tional driving force for the evolution of intense signals.
A second prediction for the evolution of acoustic
signals comes from the unusually long time-constant of
the membrane hyperpolarization. This is of the order of
5 s, and thus 1000-fold the time-constants of known
inhibitory potentials, e.g. from lateral inhibitory poten-
tials reported in neurons of the auditory pathway
(Wohlers and Huber 1978; RoÈmer et al. 1981; Selverston
et al. 1985; Wiese and Eilts 1985). Short-duration chirps
or syllables, or singing bouts of low duty cycle, would
have little or no e€ect in eliciting the suppression of
competitive signals in a€erent interneurons. In a popu-
lation of male T. viridissima competing for phonotacti-
cally responding females, long-duration singing bouts
would therefore be favoured over short ones, since
only the former would be able to reliably initiate the
190
inhibitory e€ect. Any observed choice of females for
signals with longer duration, such as in female crickets
preferring longer song bout duration (Hedrick 1986)
may thus be explained at the proximate level by passive
attraction only (Parker 1983) where females merely
move to the male producing a signal that activates its
sensory pathway most strongly.
An alternative hypothesis explains the evolution of
highly repetitive signals as a mechanism for reliable in-
formation transfer in communication systems (Wiley
1994). Highly repetitive signals with identical song ele-
ments enhance the likelihood that a receiver will avoid
evolutionary inappropriate responses (errors) in detect-
ing and/or discriminating the signal. This is particularly
helpful with a noisy transmission channel, or with
communication between widely separated individuals.
T. viridissima produces a highly redundant signal, in
which simple phonatomes are repeated at a high rate,
often for many hours of the day or night. These highly
stereotyped repetitions allow the receiving insect to
predict the entire signal when part of it is lost on the
transmission channel due to signal degradation (Wiley
and Richards 1978; RoÈmer and Lewald 1992).
Acknowledgements We thank I. Bettkes for support in calculating
the correlation coecient of action potential discharges; A. Zillmer
for logistical support in the ®eld and for collecting insects; and
H.C. Gerhardt and G. Klump for most valuable comments on an
earlier draft of the manuscript. Funding was provided by the De-
utsche Forschungsgemeinschaft through a Heisenberg Grant (HR).
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