Behavior Genetics, Vol. 16, No.

5, 1986

Behavior-Genetic Analysis of the Paradise Fish,

Macropodus opercularis. II. Passive Avoidance
Learning in Inbred Strains
Vilmos Cs~inyP and Judit Gervai ~

Received 26 July 1985--Final 12 Mar. I986

Passive dark avoidance conditioning was studied in four inbred strains

of paradise fish in the presence and absence of a fish-like dummy during
the training process. Strain differences were found in exploratory activity
during habituation trials and also in the sensitivity to the mild electric
shock punishment. The impact of the fish-like dummy also depended on
the genotype.
KEY WORDS: passive avoidance learning; strain differences; key stimuli; genotype-en-
vironment interaction.

INTRODUCTION
Passive dark avoidance conditioning of paradise fish has been studied in
previous experiments by Cs~inyi (1985b) using F1 hybrid fish of two inbred
strains, in which entrance to the dark compartment of an aquatic shuttle
box was punished by electric shock pulses. In this experiment it was also
shown that a live fish of a different species or a fish-like dummy in the
dark compartment enhanced the avoidance behavior at a low level of
shock intensity, which by itself was ineffective. By using various dummies
it was found that a dummy with two eye-like spots acted as a key stimulus,
overshadowing the less salient "darkness" signal. The important role of
eye-like key stimuli in the recognition and avoidance of a predator in this
species has also been shown in experiments with living pike (Cs~nyi,
1985a).

This work was supported by Grant 328/82 from the Hungarian Academy of Sciences.
Department of Behavior Genetics, L. E6tv6s University, G6d, Hungary 2131.
553
0001-8244/86/0900-0553505.00/0 9 1986 Plenum Publishing Corporation
554 Cs~inyi and Gervai

In the present report the passive avoidance learning of fish of different

genetic background is described in the presence and absence of a dummy,
carrying a key stimulus.

MATERIALS AND METHODS

Subjects
Female paradise fish from four inbred strains, denoted P, S, U, and
V, were used. Origin, breeding, and maintenance of these animals have
been described in detail in the first article in this series (Gervai and Cs~nyi,
1985). The strains were characterized by several marker genes (Monos-
tory et al., 1984) and by various behavioral measurements in novel sit-
uations (Gervai and Cs~nyi, 1985). The animals were tested at between
20 and 24 weeks of age.

Procedure
Forty-eight hours prior to the training experiments the fish, 24 at a
time, a total of 120, were placed in individual tanks. Passive avoidance
learning was measured using a shuttle tank described by Cs~nyi (1985b).
Shuttling activity was monitored by an infrared photoelectric device lo-
cated in the gate. This device also served as a trigger for a train of electric
shock pulses (20-msec train of 500 Hz, AC, 50 mA), if the fish entered
the dark compartment. Each animal was given two 15-rain sessions twice
daily for 5 days. The fish was placed into the light side of the apparatus
with a small hand net, and at the end of testing it was returned to its home
tank. The water in the apparatus was changed after each subject.
Sessions 1 to 6 were allowed for habituation, when entry to the dark
side was not punished. The time spent in the dark increased gradually
during habituation. Conditioning from trial 7 to trial 10 was carried out
in either the presence or the absence of a fish-like dummy. This dummy
was made from the head part of a red plastic toy goldfish, with two small
red LEDs put in place of the eyes. The dummy was placed in the back
part of the dark compartment.

RESULTS AND DISCUSSION

The results are shown in Table I.
Latency to the first entry into the dark compartment, time spent in
the dark, and number of entries to the dark side were recorded in each
session. Raw strain means and standard deviations before and during the
Behavior-Genetic Analysis of Macropodus opercularis. II. 555

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556 Csfinyi and Gervai

training sessions are given in Table I. Scaling problems were encountered

for all three measures, and In (x + 1) transformations were used to nor-
malize distributions, although within-group variances were not com-
pletely homogeneous for time spent in the dark.
The entrance latency at the end of the habituation period is somewhat
lower in strain U. Mild electric shock punishment alone resulted in in-
creased latencies in all the strains [F(4,208) = 26.64, P < 0.001] except
strain U, where the change was not significant (0.3 > P > 0.2, t test).
There was also no significant difference among the strains in the rate of
change. While the presence of the dummy greatly enhanced the increase
in latency in strain U compared with the dark signal stimulus, it had no
further significant increasing effect in strains S, V, and P. The learning
rate in the presence of the dummy was strongly genotype dependent, as
shown by the genotype-trial interaction component of variance [F(12,188)
= 4.03, P < 0.001].
Habituated mean values of time spent in the dark were high and
homogeneous across the strains, showing that all strains have a strong
preference for darkness in the shuttle box. The significant decrease in
time in the dark as a result of the shock punishment alone was found in
all four strains [F(4,208) = 47.72, P < 0.001]. It seems, though, that there
is a range of sensitivity to the mild, 50-mA shock train, with strain U
being significantly less sensitive than the other strains (P < 0.01, Duncan
multiple comparison test). This differential sensitivity is also shown by
the significant genotype-dependent difference of the rate of decrease
[F(12,208) = 2.92, P < 0.001]. However, the presence of the fish-like
dummy together with the shock punishment resulted in a further large
decrease in time spent in the dark in strain U, while a comparison of
means of control and dummy groups showed that in strains P, S, and V
there were no significant changes.
There is a considerable variation among the strains in the number of
entries to the dark compartment during the habituation period. Strains P
and V show significantly less shuttling activity than strain U, strain S
being intermediate. The shock punishment itself has a moderate decreas-
ing effect [F(4,208) = 19.37, P < 0.001] on strains P, S, and V, leaving
strain U apparently unaffected (0.2 > P > 0.1). The presence of the
dummy again results in a dramatic change in strain U, leaving the others
unchanged.
The main conclusions of this study therefore are that there is genetic
variation in the response topassive dark avoidance conditioning among
the paradise fish, and the overpowering effect of the key stimulus on the
learning process also depends substantially on the genotype. These find-
ings imply within-species variation in the interaction of species-specific
Behavior-Genetic Analysis of Macropodus opercularis. II. 557

key stimuli (SSKS) with pain or fear, which lead to a species-specific

defense reaction (SSDR) (Bolles, 1970; Cs~myi, 1985b).
It seems that among the laboratory strains P and U represent ex-
tremes, as P is a "low explorer," is sensitive to mild electric shock, and
ignores the eye-like signal stimulus, while U is a "high explorer," is less
sensitive to the electric shock, and exploits greatly the presence of the
key stimulus signal in the acquisition process. The previous study of
Csgmyi (1985b) on S x U hybrid fish showed strong directional dominance
in the direction of strain U.

ACKNOWLEDGMENT
The authors wish to thank Andrea Szebenyi for her technical
assistance.

REFERENCES
Bolles, R. C. (1970). Species-specific defence reactions and avoidance learning. Psychol.
Rev. 71:32-48.
Csfinyi, V. (1985a). Ethological analysis of predator avoidance by the paradise fish, Ma-
cropodus opercularis L. I. Recognition and learning of predators. Behaviour 92:227-
240.
Csfinyi, V. (1985b). Ethological analysis of predator avoidance by the paradise fish, Ma-
cropodus opercularis L. II. Key stimuli in avoidance learning. Anita. Learn. Behav.
14:101-109.
Gervai, J., and Csfinyi, V. (1985). Behavior-genetic analysis of the paradise fish, Macropodus
opercularis I. Characterization of the behavioral responses of inbred strains in novel
environments. A factor analysis. Behav. Genet. 15:503-519.
Monostory, Zs., Nagy, A., Gervai, J., and C s~inyi, V. (1984). Polymorphism and inheritance
of serum esterases and 13-globulins in the paradise fish, Macropodus opercularis: An-
abantidae. Anirn. Blood Groups Biochem. Genet. 15:1-11.

Edited by David Blizard