Neuroscience and Behavioral Physiology, Vol. 32, No.

4, 2002

Plasticity of Social Behavior in Drosophila

N. G. Kamyshev, G. P. Smirnova, E. A. Kamysheva, O. N. Nikiforov,

I. V. Parafenyuk, and V. V. Ponomarenko*

Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 86, No. 11, pp. 1426–1434,
November, 2000. Original article submitted April 28, 2000.

This article presents results obtained from studies of the plasticity of changes in social behavior in
Drosophila (interactions between individuals in groups) in conditions of homo- and heterogeneous envi-
ronments. This is the first report of data illustrating self-starting acquisition by female Drosophila of a
classical conditioned reflex to contextual factors signaling possible threats from other individuals and
blocking the initiation of activity. A previously described operant conditioned reflex also helped flies
avoid aggression from other individuals and make more efficient use of food resources by decreasing the
initially high level of activity. Classical conditioning had the effect that the fly did not need to repeat
acquisition of the conditioned reflex each time: when placed into an analogous situation, the fly’s activi-
ty automatically decreased as a result of exposure to the conditioned stimulus, i.e., contextual factors.

KEY WORDS: Drosophila melanogaster, learning, classical conditioned reflex, social behavior.

The interaction of two or more organisms, usually of
the same species, includes a mutual exchange of stimuli
which regulates the triggering, maintenance, and termina-
tion of the corresponding behavioral acts, and this is termed
social (community) behavior [11]. This definition includes
sexual behavior. Here, however, the discussion concerns
only those aspects of the interaction between individuals
which are not associated with sexual behavior. When
Drosophila individuals are placed in a group situation, dif-
ferent authors have observed strengthening [8–10, 14], sup-
pression [12], and the absence of any changes in activity
[13]. The contradictions in these data are evidence for the
possible involvement of previously unknown motivations,
unconditioned reflexes, and plastic changes in behavior in
controlling movement activity.
Our studies have demonstrated [1] that the direction
of changes in activity when a group situation is created
depends on whether the initial level of activity of the indi-
viduals is high or low. Initially high levels of activity
decrease, while low levels increase. One of the factors
* higher, while the probability of the transition from inactiv-
Deceased.
Laboratory for Comparative Behavioral Genetics, I. P. Pavlov
Institute of Physiology, 6 Makarov Bank, 199034 St. Peters-
burg, Russia.
401
0097-0549/02/3204-0401$27.00 ©2002 Plenum Publishing Corporation
determining differences in the level of movement activity
in individuals is the size of the experimental chamber. We
interpreted increases in activity seen in flies when space is
restricted as a Pavlovian freedom reflex [5]. The uncondi-
tioned reflex reaction whose appearance leads to increased
activity in the group situation is escape from approaching
individuals. This reaction leads to increases in the numbers
of excursions made by the flies in a group. The uncondi-
tioned reaction whose appearance leads to decreased activ-
ity in the group situation is interruption of excursions in
response to encountering another individual. This reaction
leads to decreases in the duration of excursions made by
flies in a group. Both unconditioned reactions led to a ten-
dency of flies to avoid coming into contact with each other
[9, 15] and exchanging blows [9]. The interaction of these
two reactions fixes activity and the number of contacts
dependent on activity at a relatively high level. There is,
however, a factor responsible for time-related decreases
both in the activity of flies within a group and in the fre-
quency of encounters. From the very first seconds, the
probability of the transition from activity to encounter is
ity to encounter is lower than the probability of random
movement. This creates the conditions for operant learn-
ing, whereby activity on the part of the flies leads to blows
from other individuals and inactivity allows this to be
402 Kamyshev, Smirnova, Kamysheva, Nikiforov, Parafenyuk, and Ponomarenko
Separate individuals 1 live and 9 dead
(set 1) (set 2)
70
50
30
10
A
Separate individuals Separate individuals
(set 1) (set 2)
70
50
30
10
P C1 C2 C3 C4 B P C1 C2 C3 C4 B
B
Fig. 1. The effects on the distribution of three-day Drosophila flies of the
presence of nine cold-killed flies (A) on the nutrient medium and the
absence of after-effects fro this procedure after removal of dead flies (B).
The horizontal axes show vertical zones of the flask: P = plastic tube;
C1–C4 = four zones of identical height within the flask; B = base (surface
of the medium); the vertical axes show the numbers of individuals (%).
The upper curve shows the percentage of individuals in each zone, which
is made up of two components – the percentage of immobile individuals
(shaded) and the percentage of mobile individuals (unshaded zones). Each
set was observed two times over a period of 30 min (one observation per
fly per minute): 30 min after placing live individuals with dead flies and 30
min after removal of the dead flies. Set one consisted of 110 individuals;
set 2 consisted of 102 individuals.
avoided [1]. As a result, the frequency of excursions
decreases over time and the duration of inactive periods
increases. The acquisition of an operant conditioned reflex
by flies is not accompanied by any kind of after-effect
when individuals are isolated from the group [1]. These
experiments were performed in conditions of a homoge-
neous environment – flies were placed in experimental
chambers made of a single material. After-effects, howev-
er, were seen if flies were kept in a heterogeneous environ-
ment before being placed in groups and activity recordings
on individuals taken from groups were made in the hetero-
geneous environment – standard flasks for breeding
Drosophila, with a plastic tube containing nutrient medium
on the floor [3]. This article presents previously unpub-
lished data and summarizes results from studies of plastic
changes accompanying social behavior of Drosophila in
homo- and heterogeneous environments.
METHODS
All experiments were performed using virgin females
of wild-type Canton S flies, aged three days. Flies were kept
on raisin-yeast medium at 25°C and a 12:12 light regime.
Experimental individuals were collected immediately after
hatching without any kind of immobilization. Before exper-
iments, flies were kept either singly or in groups of 10 indi-
viduals in glass flasks (diameter 25 mm, height 100 mm) on
fresh nutrient medium without live yeast. Movement activ-
ity was recorded in flasks of the same type with or without
medium, usually from 12:00 to 16:00, using a modification
of the method described by Luchnikova [4]. Vessels were
placed on a background of a white screen illuminated at an
intensity of 150 Lx and recording was started 30 min later.
The unit of observation consisted of 10 individuals kept
either in a single flask or in individual flasks. The number
of moving individuals in each observation unit were record-
ed at various time points. The index of activity (IA) for each
time period and each observation unit was calculated as
k
∑ mi
IA = i =1 ⋅ 100 %,
h⋅k
where h is the number of individuals in the observation unit;
k is the number of observations made at the given time
point; mi is number of moving individuals during the ith
observation, and i = 1 ... k. Activity indexes calculated by
this method were subjected to further statistical analysis
using parametric statistical methods (see captions to Tables
and Figures) without prior transformation of the data.
RESULTS AND DISCUSSION
In conditions of a heterogeneous environment, two
activity-suppressing factors could be added to the mecha-
nisms controlling the level of movement activity in groups.
The first appeared only when the group of flies was located
directly on the nutrient medium. Inhibition of activity
among individuals on the medium occurred even when a
group of dead individuals was placed there (Fig. 1, A;
Table 1). This activity-suppressing component did not per-
sist after individuals were transferred to another flask con-
taining medium with no other flies (Fig. 1, B). Thus, all this
unconditioned reflex activity needed was for the flies sim-
ply to perceive the presence of other individuals on the food
resource. This response reflects competitive interactions for