A combined study of behavior and Fos expression in limbic structures after re-testing Wistar rats in the elevated plus-maze.

The elevated plus-maze is an animal model used to study anxiety. In a second session, rats show a reduction in the exploratory behavior even when the two sessions are separated by intervals as large as 7 days. The aim of the present study was to investigate whether the reduction in the exploratory behavior is maintained after intervals larger than 7 days. Additionally, we aimed at investigating eventual correlations between behaviors in the plus-maze and activation of limbic structures as measured by Fos protein expression after the second session. Rats were tested for 5 min in the elevated plus-maze and re-tested 3, 9 or 33 days later. Other groups were tested only once. The rat brains were processed for immunohistochemical detection of Fos protein. The results show a decrease in the open arms exploration in the second trial with intervals of 3, 9 and 33 days. The expression of Fos protein in the piriform cortex, septal nucleus and paraventricular hypothalamic nucleus in the groups tested with intervals of 9 and 33 days were statistically different from the other groups. The alterations observed in exploratory behavior in the second session in the plus-maze did not correlate with Fos expression. In conclusion, although the specific test conditions were sufficient to evoke behavioral alterations in exploration in the elevated plus-maze, they were enough to induce significant Fos protein expression in piriform cortex, septal nucleus and thalamic and hypothalamic paraventricular nuclei but not in other areas such as dorsomedial nucleus of the hypothalamus and amygdala nuclei, known to be also active participants in circuits controlling fear and anxiety.

Thigmotactic responses in an open-field.

The goal of the present study was to investigate the role of thigmotaxis (the tendency to remain close to vertical surfaces) in rat exploratory behavior in an open-field. Thigmotaxis was investigated in a parametric way, using 24 experimentally adult naive male Wistar rats (210-230 g). Exploratory behavior was studied in an open-field (N = 12) in 5-min sessions and behavior was analyzed in terms of where it occurred: in areas surrounded by two, one, or no walls. Another group of rats (N = 12) was studied in an open-field with blocks placed near two of the corners so as to make these corner areas surrounded by three walls. The floor of the open-fields was divided into 20-cm squares in order to locate the exact place of occurrence of each behavior. The following behaviors were recorded: entries into the squares, rearings, and groomings. In both types of open-field the rats chose to remain longer in the squares surrounded by the largest possible number of walls. In one of the open-fields, the mean time (seconds) spent in squares surrounded by two walls was longer than the time spent in squares surrounded by one or no walls (37.2, 7.7, and 1.8 s, respectively). In the other open-field, the mean time spent in squares surrounded by three walls was longer than the time spent in squares surrounded by two, one or no walls (41.7, 20.4, 7.0, and 2.6 s, respectively). Other measures presented a similar profile. These results indicate that rats are sensitive to the number of walls in an environment and prefer to remain close to them.

Thigmotactic responses in an open-field

The goal of the present study was to investigate the role of thigmotaxis (the tendency to remain close to vertical surfaces) in rat exploratory behavior in an open-field. Thigmotaxis was investigated in a parametric way, using 24 experimentally adult naive male Wistar rats (210-230 g). Exploratory behavior was studied in an open-field (N = 12) in 5-min sessions and behavior was analyzed in terms of where it occurred: in areas surrounded by two, one, or no walls. Another group of rats (N = 12) was studied in an open-field with blocks placed near two of the corners so as to make these corner areas surrounded by three walls. The floor of the open-fields was divided into 20-cm squares in order to locate the exact place of occurrence of each behavior. The following behaviors were recorded: entries into the squares, rearings, and groomings. In both types of open-field the rats chose to remain longer in the squares surrounded by the largest possible number of walls. In one of the open-fields, the mean time (seconds) spent in squares surrounded by two walls was longer than the time spent in squares surrounded by one or no walls (37.2, 7.7, and 1.8 s, respectively). In the other open-field, the mean time spent in squares surrounded by three walls was longer than the time spent in squares surrounded by two, one or no walls (41.7, 20.4, 7.0, and 2.6 s, respectively). Other measures presented a similar profile. These results indicate that rats are sensitive to the number of walls in an environment and prefer to remain close to them.

Thermal stress decreases general motor activity of rats in the elevated plus-maze but does not alter aversion to the open arms.

Environmental temperature is known to affect a variety of biological processes ranging from simple chemical reactions up to ecological distribution of animal species. To our knowledge, however, there are no studies relating environmental temperature and exploration in the elevated plus-maze. The present study was aimed at investigating the influence of animal house temperature on the exploration of an elevated plus-maze. Fifty-seven male Wistar-derived rats were divided into five groups which were kept for 96-h in an animal house with different temperatures (18, 22, 26, 30 or 34 degrees C) and then tested in the elevated plus-maze. Results showed that the animals submitted to the higher temperatures decreased body weight, frequency of entries into both the open and closed arm, time spent in the open arm extremities, distance run in the closed arms and frequency of rearing while increasing the mean duration of each entry into both the open and closed arms. There were no significant effects on the time spent in the open arms and the percentage of entries into the open arms. These effect are probably due to thermal stress and do not relate to emotional changes but rather to modifications in general activity. The frequency of stretching and head-dipping exhibited different profile reactions to temperature when compared to the above measures. These behavioral modifications are also consistent with the effects of thermal stress rather than alterations in emotionality.

Effects of septal cholinergic lesion on rat exploratory behavior in an open-field.

The medial septum participates in the modulation of exploratory behavior triggered by novelty. Also, selective lesions of the cholinergic component of the septohippocampal system alter the habituation of rats to an elevated plus-maze without modifying anxiety indices. We investigated the effects of the intraseptal injection of the cholinergic immunotoxin 192 IgG-saporin (SAP) on the behavior of rats in an open-field. Thirty-nine male Wistar rats (weight: 194-230 g) were divided into three groups, non-injected controls and rats injected with either saline (0.5 microl) or SAP (237.5 ng/0.5 microl). Twelve days after surgery, the animals were placed in a square open-field (120 cm) and allowed to freely explore for 5 min. After the test, the rats were killed by decapitation and the septum, hippocampus and frontal cortex were removed and assayed for acetylcholinesterase activity. SAP increased acetylcholinesterase activity in the septum, hippocampus and frontal cortex and decreased the total distance run (9.15 +/- 1.51 m) in comparison to controls (13.49 +/- 0.91 m). The time spent in the center and at the periphery was not altered by SAP but the distance run was reduced during the first and second minutes (2.43 +/- 0.36 and 1.75 +/- 0.34 m) compared to controls (4.18 +/- 0.26 and 3.14 +/- 0.25 m). SAP-treated rats showed decreased but persistent exploration throughout the session. These results suggest that septohippocampal cholinergic mechanisms contribute to at least two critical processes, one related to the motivation to explore new environments and the other to the acquisition and storage of spatial information (i.e., spatial memory).

Effects of septal cholinergic lesion on rat exploratory behavior in an open-field

The medial septum participates in the modulation of exploratory behavior triggered by novelty. Also, selective lesions of the cholinergic component of the septohippocampal system alter the habituation of rats to an elevated plus-maze without modifying anxiety indices. We investigated the effects of the intraseptal injection of the cholinergic immunotoxin 192 IgG-saporin (SAP) on the behavior of rats in an open-field. Thirty-nine male Wistar rats (weight: 194-230 g) were divided into three groups, non-injected controls and rats injected with either saline (0.5 æl) or SAP (237.5 ng/0.5 æl). Twelve days after surgery, the animals were placed in a square open-field (120 cm) and allowed to freely explore for 5 min. After the test, the rats were killed by decapitation and the septum, hippocampus and frontal cortex were removed and assayed for acetylcholinesterase activity. SAP increased acetylcholinesterase activity in the septum, hippocampus and frontal cortex and decreased the total distance run (9.15 ± 1.51 m) in comparison to controls (13.49 ± 0.91 m). The time spent in the center and at the periphery was not altered by SAP but the distance run was reduced during the first and second minutes (2.43 ± 0.36 and 1.75 ± 0.34 m) compared to controls (4.18 ± 0.26 and 3.14 ± 0.25 m). SAP-treated rats showed decreased but persistent exploration throughout the session. These results suggest that septohippocampal cholinergic mechanisms contribute to at least two critical processes, one related to the motivation to explore new environments and the other to the acquisition and storage of spatial information (i.e., spatial memory)

Vibrissal sense is not the main sensory modality in rat exploratory behavior in the elevated plus-maze.

Four groups of male Wistar rats were submitted to acute bilateral removal of mystacial vibrissae at different lengths from the follicle. Each group was divided into two subgroups, tested under high (150 Lux) and low environmental illumination (2 Lux). All the subjects were allowed to freely explore an elevated plus-maze for 5 min. Results indicated that rats tested under low illumination tended to explore the open arms more frequently and longer then rats tested under high illumination. When tested under low illumination, rats in the group that suffered whole vibrissa removal stayed longer in the open arms than those in the other groups but did not differ in the number of entries. The average increase in the length of open arm entries, rather than a decrease in aversion to the open arms, may be due to the need of more time to obtain information about the environment since there is no light and the vibrissae were removed. This effect was not seen with rats tested under high illumination, possibly because vision could be used to obtain relevant information.

Dissociation of memory and anxiety in a repeated elevated plus maze paradigm: forebrain cholinergic mechanisms.

The effect of intraseptal injection of the cholinergic immunotoxin 192-IgG-saporin on behavior in the elevated plus maze was investigated. A 5-min test-retest paradigm, with minute-by-minute analysis of the first session, was used to evaluate both anxiety and memory in this task. Biochemical analyses revealed a decrease in acetylcholinesterase (AChE) activity in the hippocampus (HPC), septum, and frontal cortex of animals injected with IgG-192 saporin (237.5 ng) when compared with controls. No statistical differences were found between groups in terms of behaviors associated with locomotor activity, conventional measures of anxiety, or ethological behaviors during either session 1 or 2. During test session 2 the controls exhibited decreased exploratory activity and increased indices of anxiety. In contrast, the saporin-treated rats did not exhibit these experience-dependent behavioral changes from session 1 to 2. The minute-by-minute analysis showed a significant decrease in exploratory as well in anxiety associated behaviors during the first session for the control group, but not for the saporin-treated group. These results suggest that the cholinergic innervation of the HPC, the frontal cortex, or both forebrain structures, modulate the initiation of exploratory activity which, results in the acquisition and retention of spatial information, but does not affect the expression of anxiety in the elevated plus-maze.