J. exp. Biol. 130, 425-432 (1987)
Printed in Great Britain © The Company of Biologists Limited 1987
425
SHORT COMMUNICATION
A NEW MICROCOMPUTER-BASED METHOD FOR
MEASURING WALKING PHONOTAXIS IN FIELD
CRICKETS (GRYLLIDAE)
BY JOHN A. DOHERTY* AND ANTHONY PIRES
Section of Neurobiology and Behavior, Seeley G. Mudd Hall, Cornell University,
Ithaca, NY 14853, USA
Accepted 13 March 1987
Calling songs of male crickets attract sexually mature, conspecific females for
mating (for recent reviews see Eisner & Popov, 1978; Huber & Thorson, 1985;
Doherty & Hoy, 1985). This communication system has been the subject of many
behavioural studies on the relevant properties of the male's calling song for
recognition by females (e.g. Walker, 1957; Popov & Shuvalov, 1977; Pollack & Hoy,
1979, 1981; Thorson, Weber & Huber, 1982; Stout, DeHaan & McGhee, 1983;
Doherty, 19856; Doolan & Pollack, 1985). The behavioural investigations have
required some means of measuring the female's phonotaxis (her locomotion or
turning towards a sound source). These include 'closed-loop' methods in which the
animal moves freely in acoustic space and 'open-loop' methods in which the animal is
tethered and is not allowed to experience changes in sound intensities as it runs in the
direction of the sound source. Closed-loop methods include free walking in arenas
(e.g. Murphey & Zaretsky, 1972; Paul, 1976; Hoy, Hahn & Paul, 1977; Pollack &
Hoy, 1979; Stout et al. 1983); open-loop methods include tethered flight (Moiseff,
Pollack & Hoy, 1978; Pollack & Hoy, 1979, 1981; Pollack, Huber & Weber, 1984),
tethered walking on Y-maze globes (Hoy & Paul, 1973) and free walking on a
spherical locomotion compensator (Wendler, Dambach, Schmitz & Scharstein,
1980; Thorson & Huber, 1981; Thorson et al. 1982; Pollack et al. 1984; Doherty,
I985a,b,c; Schmitz, 1985).
Here we describe a new open-loop method for quantifying cricket phonotaxis.
This method uses inexpensive microcomputer technology, is completely automated
and therefore rapid and objective, and could be adapted for studying locomotory
movements of other animals. The data obtained are in such a form that they are easily
compared to data generated by the spherical locomotion compensator (Kramer
treadmill, see Kramer, 1976; Wendler^ al. 1980; Weber et al. 1981), which has been
used for quantifying phonotaxis in the field cricket, Gryllus bimaculatus (Doherty,
1985a, b,c).
•Present address: Department of Biology, Villanova University, Villanova, PA 19085, USA.
Key words: crickets, phonotaxis, communication, microcomputer.
426
J. A. DOHERTY AND A. PlRES
The spherical locomotion compensator or Kramer treadmill was first developed by
scientists in West Germany (Kramer, 1976; Wendlere* al. 1980; Weber et al. 1981;
Thorson et al. 1982). It uses an infrared detection system to monitor movements of
an untethered cricket on top of a sphere. This positional information feeds back to
servomotors that move the sphere in the opposite direction. These compensatory
movements 'fix' the cricket at the top of the sphere as it performs walking phonotaxis.
The colloquial name for this locomotion compensator is 'Kugel' (German translation
of the word 'sphere'). Because our device for measuring walking phonotaxis utilizes
the Apple Macintosh computer and is similar in appearance to the German Kugel,
we call our new device the 'MacKugeP. The main mechanical difference between
these two devices is that in the MacKugel system the cricket is tethered and its own
walking movements provide the power to rotate the sphere.
The MacKugel is a complete stimulus control and data acquisition system for online studies of cricket phonotaxis. Data on cricket movements during phonotaxis
experiments are passed directly to an Apple Macintosh microcomputer, which can
also synthesize acoustic stimuli or send control signals to an external acoustic
synthesizer. In designing this system, we took advantage of ROM routines in the
Macintosh for quantifying movements of the Apple 'mouse'.
The mouse is a mechanical, optoelectronic device that converts rotational
movements of a rubber ball to changes in the coordinates of a cursor displayed on the
computer's CRT monitor. The ball bears against the operator's desk surface, and
against three rollers inside the ball's housing. One roller is for mechanical support
only. The other two, representing orthogonal x- and y-axes, are connected to rotating
vanes which interrupt light beams between LED—phototransistor pairs. The
phototransistors produce electronic signals which are sent to the computer.
We adapted the mouse to study cricket phonotaxis by expanding the distance
between the rollers and replacing the small rubber ball (2-5 cm diameter) with a
larger and much lighter sponge-rubber ball (10 cm diameter, 8-7g). This larger ball
(a hollowed-out, toy 'Nerf ball' from Parker Bros, Beverly, MA, USA) was mounted
in a frame along with the roller/phototransistor modules from the mouse (Fig. 1). In
this way the rollers could be actuated by a tethered cricket walking on top of the ball.
As the cricket ran in one direction (i.e. towards an attractive acoustic stimulus), the
ball was moved in the opposite direction and the movements were transduced into
changing pixel coordinates on the Macintosh screen. The minimum force required to
move the Nerf ball ranged from 1-117 to 2-793xlO~2N. A plastic sphere (10 cm
diameter, 12-8 g) is currently under development and this improved version only
requires forces ranging from 0-69 to 1-05x10" N. High-level computer languages
(Macintosh Pascal®) were used for data acquisition and analysis (program available
from author). By sampling pixel coordinates once every second, we were able to
calculate instantaneous velocity and direction profiles for the cricket's movements.
These profiles, along with their corresponding 'vector plots' (see below), were
displayed simultaneously on the Macintosh screen, and the data were saved on
micro-floppy disks for subsequent data analysis.
Measurement of phonotaxis in crickets
RT
427
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Fig. 1. Side (A) and top (B) view of MacKugel device. TM, tether mount; F, frame;
RT, rotating tether; PR, passive roller; T,, x-axis tachometer (composed of phototransistor module, extension axle and roller); T y , y-axis tachometer; FB, frictionless
bearing (composed of a 1 cm Teflon ball riding on a film of air); AS, air supply (air routed
through a threaded aluminium cylinder used to adjust the height of the Nerf ball);
S, sphere.
To establish the usefulness and validity of our method, we repeated some
locomotion compensator experiments run previously on the field cricket, G. bimaculatus (Doherty, 1985a,b). In both methods, walking phonotaxis in crickets can be
quantified by measuring time profiles of the cricket's walking velocity and direction.
Fig. 2 shows these time profiles for one MacKugel experiment in which a synthetic
calling song was presented to a female G. bimaculatus from one of two loudspeakers
separated by 180°. This calling song was played from the left speaker for 2min and
then from the right speaker for another 2min. A 30-s silent period preceded and
followed this playback. The female tracked the stimulus, as shown by her narrow
meandering about the direction of the active speaker. Her walking velocity also
increased when the stimulus was presented, compared to the preceding silent period.
The time profiles shown in Fig. 2 were generated by sampling and storing the
position of the animal once each second during an experiment. This method of data
collection easily lent itself to another way of representing phonotactic movements
visually. For each 1-s interval, a vector was calculated that showed the animal's
walking direction relative to the direction of the active speaker (vector angle) and its
velocity (vector length). These vectors were cumulatively plotted as shown in Fig. 2.
These 'vector plots' show that when the stimulus was presented from either the left or
right speaker, the vectors were clustered about the direction of the active speaker.
428
J. A. DOHERTY AND A. PlRES
The direction component of cricket walking phonotaxis is a sensitive indicator of
the attractiveness of an acoustic stimulus. To investigate this sensitivity further, we
ran phonotaxis experiments in which acoustic stimuli with different pulse periods
were presented in a sequential, to-and-fro paradigm. As in earlier experiments of
cricket phonotaxis on a locomotion compensator (Doherty, 1985c), pulse periods
ranging from 30 to 50 ms yielded the best tracking on the MacKugel (i.e. best
orientation to active speaker direction, see Fig. 3).
CP
PD
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Angular d
1 RS
LS
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1 min
360'
Fig. 2. Phonotaxis of a female Gryllus bimaculatus on the MacKugel device. A synthetic
calling song composed of 4-pulse chirps repeated at a constant rate (40 ms pulse period,
PP; 4 pulses per chirp, PN; 22 ms pulse duration, PD; 400 ms chirp period, CP; shown in
box) was presented from one of two speakers for 4 min. The cricket's movements were
translated into cursor movements on the CRT of a microcomputer. The cursor's position
(in x—y pixel coordinates) was sampled once every second and vector plots and time
profiles of the cricket's velocity and angular direction were calculated and plotted during
the experiment. The two vector plots show the accumulation of vectors during stimulus
playbacks from the two loudspeakers. Each vector has an angle and a length. Vector angle
is the direction the animal was moving during the sample interval, relative to the position
of the active speaker. Vector length is directly proportional to the animal's velocity. Every
vector was plotted with its origin at the centre of the vector plot circle. The radius of this
circle corresponds to a velocity of 3-4cms~'. The solid dot beside each vector plot
indicates the position of the active speaker (left or right speaker; LS, RS), which
corresponds to the active speaker position (serrated horizontal line at either 90° or 270°)
in the angular direction profile below. The numbers above each vector plot are the vector
score (left-hand number, defined in text) and the percentage of the stimulus presentation
time that the animal was moving (right-hand number). The vector score (2426) and the
percentage of time spent moving (97 %) were both higher when the stimulus was played
from the right speaker.
Measurement of phonotaxis in crickets
429
In these same phonotaxis experiments on the MacKugel we found slight effects of
pulse period on the walking velocity profiles. Females walked for a greater percentage
of the stimulus presentation time when pulse periods were optimal (i.e. 30-50ms)
0-6 -i
40
39
0-4-
0-2-
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O
00
-0-2
10
20
30
40
50
60
Pulse period (ms)
70
80
Fig. 3. Effects of synthetic calling songs with different pulse periods on the accuracy of
orientation to the speaker. Orientation was quantified as follows: for each 1-s interval of a
trial in which the animal was moving, the cosine of the vector angle was calculated. A
mean cos (angle) was then obtained for all such intervals within a trial. An average
cos (angle) was computed for all trials of each pulse period and plotted as orientation to
speaker. If the cricket ran directly towards the loudspeaker (defined as 0°), the cos (angle)
was 1; this value was — 1 when it ran directly away from the speaker and 0 when it ran
perpendicular to the sound source. The points are means for each pulse period and the
vertical lines are standard errors. Data were based on to-and-fro scans of pulse period for
21 female Gryllus bimaculatus. Sample sizes are shown.
600-1 B
70-i
i -100
20
30
40
50
60
70
80
10
20
30
40
50
60
70
Pulse period (ms)
Fig. 4. Comparison of two different methods of quantifying stimulus efficacy in
sequential, to-and-fro scans of pulse period (see text and Doherty, 1985a,c). The two
methods were (A) manual measurements of the percentage of stimulus presentation time
that females tracked the stimulus and (B) microcomputer measurements of vector score.
The criteria for manual scoring of stimulus tracking are found in Doherty (1985a,c). The
points are mean values and the vertical lines are standard errors. Nine Gryllus
bimaculatus females were tested. Sample sizes are the same in A and B.
80
430
J. A. DOHERTY AND A. PlRES
and walked less in response to stimuli with pulse periods outside this range.
Furthermore, mean walking velocity was highest in response to pulse periods of 35
and 40 ms, whether or not pauses in walking were included in the calculation.
Because both walking velocity and direction were affected by acoustic stimuli, we
devised a numerical 'vector score' which incorporates both of these components and
serves as a quick measure of the phonotactic efficacy of different sound stimuli. This
score is defined as:
Vector score = 2[cos(vector angle) X vector length].
In calculating the vector score, the angular direction of the active speaker is always
defined as 0° and the summation is over all the sample intervals within a trial. When
the animal runs towards the loudspeaker the vector score increases, when it runs
away from the loudspeaker the score decreases. By definition, movements perpendicular to the speaker axis (cos 90° or cos 270°) and no movements at all (vector
length = 0) result in vector scores approaching zero. For example, in Fig. 2, the
vector score in response to calling song from the left speaker was lower than that for
the right speaker. This difference was due to less accurate tracking, wider
meanderings about the speaker direction and more pauses in walking.
The vector score is comparable to other measures of stimulus efficacy in cricket
phonotaxis. In earlier studies of G. bimaculatus phonotaxis on a locomotion
compensator, the attractiveness or efficacy of an auditory stimulus was quantified by
measuring the percentage of the stimulus presentation time that the female clearly
'tracked' the stimulus. Tracking has been defined as meandering within a certain
angle 'window' (±60°) about the angular direction of the active speaker (see Thorson
et al. 1982; Doherty, 1985a,b,c). These same criteria were used in our study for
measuring the percentage of time spent tracking in a group of nine G. bimaculatus
females. In to-and-fro sequential experiments, tracking scores and vector scores in
response to calling songs with different pulse periods were comparable (Fig. 4). Both
scores showed that songs with pulse periods between 35 and 50 ms were most
effective in eliciting positive phonotaxis.
The MacKugel system is also useful for quantifying cricket phonotaxis in twostimulus (choice) playback experiments. The total vector score was a sensitive,
objective measure of the relative attractiveness of alternative acoustic stimuli. When
given a choice between alternating chirps with different pulse periods (PP), female
G. bimaculatus clearly preferred the standard chirp stimulus (40 ms PP) to the
alternative chirp stimuli with pulse periods of 20, 30, 50 and 60 ms. The strength of
the female's preference for the standard over the alternative pulse period was also
reflected in the mean vector score. These results were consistent with those of
previous choice experiments on a locomotion compensator (Doherty, 1985c).
The MacKugel system is an efficient, rapid and objective method for measuring
the efficacy of different acoustic stimuli in eliciting walking phonotaxis of crickets.
Using this system the data can be displayed to give a qualitative, visual representation of an individual cricket's movements. Complicated time profiles of movement
velocity and direction can be reduced objectively to a numerical score that can be
Measurement of phonotaxis in crickets
431
used as a rough basis of comparison between individuals and between experimental
treatments. Data analysis can be done 'on-line' as the animal is behaving. Other
strengths of the MacKugel are its low price, simplicity of design, and efficient
implementation of existing microcomputer technology.
This research was supported by grants from NSF and NIH to JAD, AP and
Ronald R. Hoy. We thank Alan Cohen and Rick Nicoletti for technical assistance and
Wendy Sussdorf for preparing Fig. 1. Ronald Hoy, Peter Brodfuehrer and Jud
Crawford read earlier drafts of this manuscript.
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