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Fig. 1. A time chart of animal preparation and experiments. Pairs of animals share
aquaria (rectangular boxes), where they can be isolated from each other by an
opaque barrier (gray vertical bar), kept apart by an open divider that allowed
pairs to smell, see and touch each other but not to fight (broken gray bar),
or interact freely (gray double-headed arrow). The sequence of treatments is
shown vertically, in rows labeled from A to K. Tests are indicated by light
coloring of the rectangles. Animals were divided into two groups, Isolates and
Social Group. (A) All animals were separated from an aquarium partner by a
closed divider (i.e. isolated) for four weeks. (B) (two rows) Isolates were
separated for two more weeks and thereafter through the tests. Social pairs
were allowed to interact freely and form dominance relationships for 30 min
day–1; at other times they were kept apart by an open
divider. Dominance status was determined during each 30 min of interaction by
scoring the numbers of aggressive and submissive behaviors performed by each
member of a pair (see equation in Materials and methods and
Fig. 3). (C) At the end of the
two weeks, the social pairs were re-isolated overnight (
15 h) to prevent
further agonistic interactions. (D) On experimental Day 1, Isolates and the
socially paired animals were tested while separated. (E) Isolates and socially
paired animals remained separated overnight. (F) On Day 2, Isolates remained
separated while social pairs were allowed to interact freely with their
partners for 30 min. (G) All animals were tested, Isolates while separated and
social pairs while in the presence of their partner. (H) Social pairs were
kept apart by an open divider for 30 min and then (I) tested over a 2–3
h period. (J) Social pairs were then re-isolated by a closed divider for 30
min and then (K) tested again over 2–3 h while isolated.
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