HIPPOCAMPUS 15:260 –272 (2005)

Hippocampus and Remote Spatial Memory in Rats

Robert E. Clark,1,2* Nicola J. Broadbent,2 and Larry R. Squire1,2,3,4

ABSTRACT: Damage to the hippocampus typically produces tempo- taken to support the idea that the structures damaged in
rally graded retrograde amnesia, whereby memories acquired recently are amnesia have a temporary role in the formation and stor-
impaired more than memories acquired remotely. This phenomenon has
been demonstrated repeatedly in a variety of species and tasks. It has also age of memory (Squire and Alvarez, 1995). Formal stud-
figured prominently in theoretical treatments of memory and hippocam- ies have clarified the phenomenon considerably. Thus,
pal function. Yet temporally graded retrograde amnesia has not been retrograde amnesia tends to be temporally graded when
demonstrated following hippocampal damage in spatial tasks like the damage is limited to the hippocampal region (Manns et
water maze. We have assessed recent and remote spatial memory follow- al., 2003; Kapur and Brooks, 1999) and extensive and
ing hippocampal lesions in three different tests of spatial memory: (1) the
standard water maze; (2) the Oasis maze, a dry-land version of the water ungraded when the damage includes lateral temporal
maze; and (3) the annular water maze, where training and testing occur neocortex (Kapur, 1993; Squire and Alvarez, 1995).
within a circular corridor. Training protocols were developed for each During the 1990s, retrograde amnesia began to be stud-
task such that retention of spatial memory could be expressed after very ied prospectively in experimental animals. Through 2003,
long retention intervals. In addition, retention in each task was assessed
with single probe trials so that the assessment of remote memory did not one can identify as many as 20 studies in which animals were
depend on the ability to relearn across multiple trials. The findings were given equivalent amounts of training on two or more occa-
consistent across the three tasks. In the standard water maze (Experiment sions before damage to the hippocampus, fornix, or ento-
1), spatial memory was impaired after training–surgery intervals of 1 day, rhinal cortex. Of these, 16 studies found temporally graded
8 weeks, or 14 weeks. Similarly, in the Oasis maze (Experiment 2), spatial
retrograde amnesia. The typical extent of the retrograde am-
memory was impaired after training–surgery intervals of 1 day and
9 weeks. Finally, in the annular water maze (Experiment 3), spatial mem- nesia gradient was about 30 days (Squire et al., 2005). In-
ory was impaired after training–surgery intervals of 9 weeks and terestingly, in the four cases in which a temporal gradient of
14 weeks. Dorsal hippocampal lesions impaired performance to the same retrograde amnesia was not observed following hippocam-
extent as complete lesions. The impairment in remote spatial memory pal damage, three involved the water maze (Bolhuis et al.,
could reflect disruption of previously acquired spatial information. Alter-
natively, it is possible that in these tasks hippocampal lesions might 1994; Mumby et al., 1999; Sutherland et al., 2001). In these
produce an impairment in performance that prevents the expression of an three studies, memory was impaired similarly regardless of
otherwise intact spatial memory. Published 2004 Wiley-Liss, Inc.† the interval between training and surgery. The water maze
requires the animal to swim to a specific point in space, using
KEY WORDS: water maze; annular maze; retrograde; Oasis maze
distal cues as guides. Studies with the water maze can be
contrasted with three others that also required the animal to
INTRODUCTION use spatial information, but only to discriminate between
two (Cho et al., 1993; Cho et al., 1995) or three (Ramos,
Descriptions of human memory impairment have long emphasized that 1998) arms of a maze rather than to find a specific location.
remote memory is typically spared relative to recent memory (Ribot, 1881). In these latter studies, temporal gradients of retrograde am-
This phenomenon, termed temporally graded retrograde amnesia, has been nesia were observed, that is, remote memory was relatively
preserved.
It is unclear why remote memory was impaired by
1
Veterans Affairs Medical Center, San Diego, California; 2Department of hippocampal lesions in a task that requires the animal to
Psychiatry, University of California, San Diego, La Jolla, California; 3De- remember a specific point in space. One possibility is that
partment of Neurosciences, University of California, San Diego, La Jolla, this type of spatial memory always requires the hip-
California; 4Department of Psychology, University of California, La Jolla, pocampus because the hippocampus forms and stores the
California
Grant sponsor: Medical Research Service of the Department of Veterans
essential spatial maps needed for task performance
Affairs; Grant sponsor: National Institute of Mental Health (NIMH); Grant (O’Keefe and Nadel, 1978). Alternatively, it is possible
number: 24600; Grant sponsor: Metropolitan Life Foundation; Grant spon- that a hippocampal lesion interferes with the ability to
sor: National Institute of Aging; Grant number: P50 AG05131; Grant perform the spatial task, rather than the ability to remem-
sponsor: National Science Foundation (NSF); Grant number: 0237053; ber a specific location. Thus, the lesion might impair
Grant sponsor: James S. McDonnell Foundation; Grant number: 21002077;
Grant sponsor: National Alliance for Research on Schizophrenia and De- navigational abilities, or the lesion might impair new
pression Effie Beeman Investigator Award. learning that is required so that the animal can update its
*Correspondence to: Robert E. Clark, Department of Psychiatry, Room position in space during the performance test (Knowlton
0603, UCSD La Jolla, CA 92093. E-mail: reclark@ucsd.edu and Fanselow, 1998). One should also consider the pos-
Accepted for publication 15 September 2004
DOI 10.1002/hipo.20056 sibility that some peculiarity of the water maze task itself
Published online 2 November 2004 in Wiley InterScience (www. underlies the finding that remote spatial memory was
interscience.wiley.com). impaired by hippocampal lesions. In this respect, it is

Published 2004 WILEY-LISS, INC. †This article is a US government work and, as such, is
in the public domain in the United States of America.
____________________________________ HIPPOCAMPUS AND REMOTE SPATIAL MEMORY IN RATS 261

notable that some sparing of remote spatial memory was reported

in a dry land version of the water maze task (Kubie et al., 1999).
It is also notable that patient E.P., who developed profound
amnesia at the age of 70 following extensive bilateral damage to the
medial temporal lobe, demonstrated intact spatial memory for the
neighborhood in which he grew up (Teng and Squire, 1999).
Perhaps spatial memory requires quite a long time to become in-
dependent of the hippocampus. Interestingly, it has been difficult
to assess spatial memory in rodents after very long retention inter-
vals, because intact animals often do not remember for longer than FIGURE 1. Top view of the three spatial tasks. All three tasks
a few weeks (e.g., Sutherland et al., 2001), or they perform near require the use of distal spatial cues (e.g., posters located on testing
chance levels (e.g., near 30% when chance ⫽ 25%; Bolhuis et al., room walls). A: Water maze (Experiment 1). Rats were trained to find
an escape platform hidden just below the surface of the water. B: Oasis
1994). maze (Experiment 2). Water-deprived rats were trained to find a sin-
These considerations suggest that the time is ripe for a thorough gle well containing a 0.3 ml drop of water among 426 wells. C: An-
examination of the effects of hippocampal lesions on recent and nular water maze (Experiment 3). Rats were trained to find an escape
remote spatial memory. Accordingly, we have assessed spatial platform hidden just below the surface of the water. The swim path
was restricted by clear walls that formed an annulus, such that rats
memory after long retention intervals in each of three different
necessarily encountered the platform location in the course of swim-
spatial memory tasks: (1) the water maze, which has been the ming. Recognition of the platform location during probe trials was
benchmark task of spatial memory in the rat (Experiment 1); (2) indicated by a slowing of swim speed in the vicinity of the platform.
the Oasis maze, which is a dry land version of the water maze task Scale bar ⴝ 1 m for all three mazes.
(Experiment 2); and (3) the annular water maze, which is a task of
spatial recognition memory that removes the need for spatial nav-
igation (Experiment 3).
Apparatus
We gave animals extensive training to improve the long-term Water maze testing was conducted in a pool of water (1.8-m
retention of spatial memory and also to provide a long period diameter at the water level) that was rendered opaque by the addi-
during which memories might become independent of the hip- tion of powdered milk. The room was illuminated by four 30-W
pocampus. Further, the entire hippocampus was removed at dif- spotlights pointed at a white ceiling. The water was maintained at
ferent times after training so that results would not be complicated room temperature. The testing room contained a number of con-
by sparing of hippocampal tissue. Finally, retention of spatial stant, salient visual cues (posters, objects, and equipment). A video
memory was assessed with probe trials, in order to obtain a direct camera was mounted on the ceiling directly above the pool and was
measure of memory that was not confounded by the difficulty used in conjunction with a video tracking system (San Diego In-
animals would be expected to have in relearning the tasks. struments) to record the swim path of each rat.
We used an Atlantis platform which could be raised or lowered
remotely (Spooner et al., 1994). When the platform (12.7-cm
EXPERIMENT 1: WATER MAZE diameter) was in the lowered position, the rat could neither detect
the platform nor escape from the water. When the platform was in
In the water maze, a rat learns to escape water by using spatial the raised position (1.5 cm below the surface of the water), it
cues to locate a platform hidden just below the water surface (Mor- remained invisible to the rat but provided a means to escape the
ris, 1984) (Fig. 1A). This task has proved to be quite sensitive to water. The Atlantis platform provides two advantages over the
hippocampal damage, and even a partial lesion is sufficient to standard, static platform. First, it provides the opportunity to
impair performance (e.g., Moser et al., 1993, 1995; de Hoz et al., present reinforced probe trials. That is, a probe trial can be pre-
2003; Broadbent et al., 2004). We developed a training protocol sented (to assess retention) with the platform in the lowered posi-
that would allow spatial memory to be expressed on a single probe tion. When the probe trial ends, the platform is raised so that the
trial even after long retention intervals. We then compared the rat can escape and be reinforced for searching in the correct loca-
performance of control animals and animals with large hippocam- tion. Second, the rats can be shaped to “dwell” over the correct
pal lesions made 1 day, 8 weeks, or 14 weeks following training. platform location. This procedure trains the rat to be both accurate
For comparison, an additional group was included that received and persistent and it substantially improves the retention perfor-
dorsal hippocampal lesions 8 weeks after training. mance of normal rats on probe trials (i.e., it increases the percent-
age of time spent in the correct location; Spooner et al., 1994).

Subjects Behavioral Training

We tested 73 experimentally naive, male Long-Evans rats
Phase I. Platform training
weighing 300 –350 g at the beginning of the experiment. Rats were
housed in pairs and maintained on a 12:12-h light/dark cycle. All During this phase of training, curtains surrounded the pool and
rats had free access to food and water. blocked the distal spatial cues. An object was suspended 20 cm
262 CLARK ET AL.

above the hidden platform and served as a beacon for the rats to For the rats given complete hippocampal lesions (H-RF), sur-
find the platform. To begin each trial, the rat was placed in the gery was scheduled 1 day (n ⫽ 8; control group, n ⫽ 8), 8 weeks
water, facing the pool wall, at one of 4 start points around the pool (n ⫽ 9; control group, n ⫽ 16), or 14 weeks (n ⫽ 12; control
(i.e., North, South, East, West). Two trials were given each day group, n ⫽ 12) after training. For rats given dorsal hippocampal
from each start point (eight trials/day for 3 days). The hidden lesions (DH-RF), surgery was scheduled 8 weeks after training
platform was raised to within 1.5 cm of the water surface only (n ⫽ 8).
when the rat was both accurate (⬍20 cm from the platform) and At the completion of behavioral testing, rats were administered
persistent in searching at the correct location (dwell time of 0.5 s an overdose of sodium pentobarbital and perfused transcardially
on the first day, 1.5 s on the second day, and 2.5 s on the third day). with buffered 0.9% NaCl solution followed by 10% formaldehyde
After escaping, the rats remained on the platform for 30 s before (in 0.1 M phosphate buffer). The brains were then removed and
being removed. If the rat failed to find the platform within 90 s, it cryoprotected in 20% glycerol/10% formaldehyde. Coronal sec-
was guided to the platform where it remained for 30 s. This pro- tions (50 ␮m) were cut with a freezing microtome. Every fifth
cedure shaped a focused and sustained search at the platform loca- section was mounted and stained with thionin to assess the extent
tion. of the lesions.

Phase II. Spatial training Phase III. Retention probe

Following platform training, the curtains were removed to re- Three 60-s probe trials were administered 14 days after surgery.
veal the distal spatial cues. Rats were given eight trials each day for The trials began by placing the rat in the water, facing the pool
10 days (intertrial interval approximately 8 min; 2 trials/day from wall, at one of the 4 start points (counterbalanced within groups).
each of the starting points and with the sequence of starting points The platform remained lowered for 60 s before being raised to
counterbalanced within groups). The platform location was the provide escape. Each rat remained on the platform for 30 s before
same for all animals. The platform was raised to permit escape from being removed. Performance on the probe trials was calculated by
the water when the rat dwelled within 20 cm of the platform for measuring the percentage of time that each rat spent in the quad-
2.5 s. After escaping, it remained on the platform for 30 s. The first rant of the pool where the platform had been located during train-
and fifth trial of each training day were 30-s, reinforced probe ing (chance performance ⫽ 25%). We also used an alternative
trials. During these trials, the platform was in the lowered position. method for analyzing probe trial performance (Moser et al., 1993)
After 30 s the platform was raised. This procedure allowed spatial by calculating the percentage of time that a rat spent in a circular
learning to be tracked during the 10 training days. zone (30 cm diameter) around the point where the platform had
been located during training. Chance performance ⫽ 4% (i.e., a
Surgery and histology. Anesthesia was maintained throughout 30-cm circle represents 4% of the total area of the pool).
surgery with isoflurane gas (0.8 –2.0% isoflurane delivered in O2 at
1 L/min). The animals were placed in a Kopf stereotaxic instru-
ment, and the incisor bar was adjusted so that bregma was level Results
with lambda. Thermocoagulation lesions were made with a radio-
frequency electrode and generator (Radionics model RF-4A). The Histological findings
electrode was first lowered to the surface of the dura, and a small
puncture was made in the dura just below the electrode tip. The Figure 2A illustrates the extent of the largest and smallest lesion
electrode was then lowered to the target and left in place for 1 min in each group.
before heating the tissue to 80 –90°C (depending on the target site)
for a period of 1 min. The current to the electrode was then turned Hippocampal damage H-RF groups. All animals sustained
off, and the electrode was removed after the tip temperature fell to extensive bilateral damage to all the cell fields of the hippocampus,
41°C (see Clark et al., 2000 for additional details). Lesions were including the dentate gyrus. The average percentage damage to the
intended to damage the dorsal and ventral hippocampus (H-RF) hippocampus was 85.3% (range 74.5–91.0%). The primary areas
or only the dorsal hippocampus (DH-RF). Lesions were made at of spared hippocampal tissue were located along the midline of the
multiple locations: All coordinates are in millimeters and relative dorsal hippocampus and involved primarily the most medial as-
to bregma (leveled to lambda). AP ⫺2.4, ML ⫾ 1.0, DV ⫺3.5; AP pects of CA1 and the crest of the dentate gyrus. All rats had sparing
⫺3.2, ML ⫾ 1.4, DV ⫺2.7; AP ⫺3.2, ML ⫾ 3.0, DV ⫺2.7; AP of the most ventromedial portion of the ventral hippocampus.
⫺4.0, ML ⫾ 2.5, DV ⫺2.3; AP ⫺4.0, ML ⫾ 3.7, DV ⫺2.7; AP Occasionally, small islands of spared hippocampal tissue were ob-
⫺4.8, ML ⫾ 4.9, DV ⫺6.8*; AP ⫺4.8, ML ⫾ 4.3, DV ⫺7.4*, served within the damaged areas of the hippocampus. All animals
⫺3.5; AP ⫺5.4, ML ⫾ 4.2, DV ⫺4.2*; AP ⫺5.4, ML ⫾ 5.0, DV had some damage to the alveus and to the fimbria on the dorsal
⫺6.5*, ⫺5.5*, ⫺4.5*. Asterisks indicate those locations that were edge of the dorsal hippocampus. Most animals sustained at least
omitted when the surgery was designed to damage only the dorsal minor damage to the dorsal and ventral subiculum. When present,
hippocampus. The wounds were then closed, and the rats recov- the damage tended to occur in the ventral subiculum just below the
ered from anesthesia on a water-circulating heating pad. All ani- ventral hippocampus. The large portion of the subiculum posterior
mals were allowed to recover for 14 days before behavioral testing. to the ventral hippocampus was spared.
____________________________________ HIPPOCAMPUS AND REMOTE SPATIAL MEMORY IN RATS 263

FIGURE 2. Reconstructions of coronal sections showing the largest (striped) and smallest
(black) area of damage for rats with large hippocampal lesions (H-RF) and dorsal hippocampal
lesions (DH-RF). Reconstructions from rats in Experiment 1(A), Experiment 2 (B), and
Experiment 3 (C). Each series of sections progresses (top to bottom) from anterior to posterior
levels. Numbers represent the distance in millimeters posterior to bregma.

Other damage. Three animals sustained minor damage to the (25% and 4% respectively; t ⬎ 6.7, P ⬍ 0.0001). Based on these
entorhinal cortex that amounted to less than 10% of total entorhi- scores, the rats were divided into equivalent lesion and control
nal volume. The entorhinal cortex was entirely spared in all other groups.
animals. In most cases, there was some detectable damage to the
cortical regions directly dorsal to the dorsal hippocampus. Most Initial retention probe: quadrant analysis. Figure 3 illus-
frequently this damage was in the form of cortical thinning, but a trates the percentage of time that each group spent in the training
few cases involved direct tissue damage. No animal had any dam- quadrant during the initial 60-s retention probe trial (chance ⫽
age to the perirhinal cortex or the amygdala. 25%). For the three control groups, the scores were 67.6 ⫾ 5.2%,
50.9 ⫾ 5.6%, and 31.2 ⫾ 6.7% for the 1-day, 8-week, and
Hippocampal damage DH-RF group. All animals sustained 14-week intervals, respectively. These scores were above chance for
extensive bilateral damage to all the cell fields of the dorsal hip- the 1-day and 8-week groups (t ⬎ 4.5, P ⬍ 0.001), but not for the
pocampus, including the dentate gyrus. The average percentage 14-week group (t[11] ⫽ 0.9, P ⬎ 0.10). Thus, forgetting was
damage to the hippocampus was 37.9% (range 31.3– 41.7%). The evident in the control group as time passed after training (one-way
primary areas of spared hippocampal tissue were located along the analysis of variance [ANOVA], (F[2] ⫽ 7.2, P ⬍ 0 .01).
midline of the dorsal hippocampus and involved primarily the By contrast, rats with hippocampal lesions performed similarly
most medial aspects of CA1 and the crest of the dentate gyrus. All to each other (t ⬍ 0.7, P ⬎ 0.10) and at chance at all training–
rats had sparing of the ventral hippocampus and the ventral sub- surgery intervals (t ⬍ 1.9, P ⬎ 0.09). The scores were 30.2 ⫾
iculum. Several of the animals had detectable damage to the alveus 4.2%, 29.6 ⫾ 2.5%, and 26.1⫾ 4.7% for the 1-day, 8-week, and
and to the fimbria. 14-week H-RF groups, respectively (chance ⫽ 25%), and the score
for the DH-RF group was 27.0% ⫾4.5%. The 1-day H-RF group
Other damage. There was no substantial extrahippocampal performed more poorly than the 1-day control group (t[14] ⫽ 5.6,
damage in this group, and no cortical thinning as noted in the P ⬍ 0.0001), and both the H-RF and DH-RF groups at 8 weeks
H-RF groups. performed more poorly than the 8-week control group (t ⬎ 2.7,
P ⬍ 0.05).
Behavioral Findings
Acquisition. All rats readily learned the platform location Initial retention probe: circle analysis. The results were the
within the first few days of training. During the first probe trial on same as in the quadrant analysis when retention was measured by
the tenth and final day of training, rats spent 50.4 ⫾ 3.8% of the the percentage of time that rats spent in a small circle around
time in the target quadrant and 24.3 ⫾ 2.6% in the circle around the platform location (chance ⫽ 4.0%). For the three control
the platform. Both of these scores were well above chance levels groups, the scores were 41.2 ⫾ 7.1%, 21.1 ⫾ 3.8%, and 8.2 ⫾
264 CLARK ET AL.

39.9 ⫾ 4.6%, respectively; t ⫽ 1.3, P ⬎ 0.1). On the third probe

trial, the control groups continued to increase their percentage
time in the target quadrant (all scores ⬎70%). However, among
the lesion groups, only the 8-week DH-RF group remained above
chance (47.8 ⫾ 5.6%; t ⫽ 4.1, P ⬍ 0.01). For this probe trial the
14-week H-RF group spent only 33.6 ⫾ 6.1% in the target quad-
rant.

Retention probes 2 and 3: circle analysis. The findings from

the circle analysis were the same as for the quadrant analysis with
the following exception: The 14-week H-RF group did improve on
probe trial 2 (from 6.9 ⫾ 2.1% to 11.3 ⫾ 2.8%; t[11] ⫽ 2.1, P ⫽
0.06) but was still impaired relative to the 14-week control group
on the second probe trial (26.7 ⫾ 5.9%; t[24] ⫽ 2.4, P ⬍ 0.05).

Discussion
The control groups exhibited forgetting as the retention interval
lengthened, and there was scant evidence of retention in the
FIGURE 3. Water maze performance (Experiment 1). Percentage 14-week group. Performance of the lesion groups was identical on
time in the training quadrant during a retention probe given 14 days the initial probe trials irrespective of the retention interval. Fur-
after surgery. Control rats (CON), rats with complete hippocampal ther, none of the lesion groups exhibited evidence of memory
lesions (H-RF), and rats with dorsal hippocampal lesions (DH-RF) retention, either by an analysis of percentage time in the target
were tested for retention after a 1-day (CON, n ⴝ 8; H-RF, n ⴝ 8), quadrant (Fig. 3) or by an analysis of percentage time in a small
8-week (CON, n ⴝ 16; H-RF, n ⴝ 9; DH-RF, n ⴝ 8), or 14-week
(CON, n ⴝ 12; H-RF, n ⴝ 12) training–surgery interval. None of the circle around the target.
lesion groups performed above chance (25%). The 1-day and 8-week On the second probe trial, the performance of all of the control
control groups performed above chance and better than the corre- groups improved, as did the performance of the DH-RF 8-week
sponding lesion group. Error bars ⴝ SEM. group and the H-RF 14-week group. The good performance of the
14-week H-RF group on probe trial 2 could be evidence of a
2.5% for the 1-day, 8-week, and 14-week intervals, respectively. reminder effect from the first probe trial that built on a weak spatial
These values were above chance for the 1-day and 8-week groups memory. If so, some spatial memory was present in this group, but
(t ⬎ 4.4, P ⬍ 0.01), but not for the 14-week group (t[11] ⫽ 1.6, not in the other groups where large hippocampal lesions were made
P ⫽ 0.13). By contrast, the lesion groups performed similarly to sooner after training. However, it is also notable that performance
each other (t ⬍ 1.5, P ⬎ 0.10) and at chance at all training–surgery of the 14-week H-RF group declined to chance on the third probe
intervals (t ⬍ 1.5, P ⬎ 0.10). The scores were 6.0 ⫾ 3.0%, 4.7 ⫾ trial.
2.0%, and 6.9 ⫾ 2.1% for the 1-day, 8-week, and 14-week H-RF
groups, respectively, and the score for the DH-RF group was 4.0%
⫾ 1.2%. The 1-day H-RF group performed more poorly than the EXPERIMENT 2: OASIS MAZE
1-day control group (t[14] ⫽ 4.6, P ⬍ 0.001), and the H-RF and
DH-RF groups at 8 weeks performed more poorly than the 8-week To date, previous studies using the water maze (Bolhuis et al.,
control group (t ⬎ 3.0, P ⬍ 0.01). 1994; Mumby et al., 1999; Sutherland et al., 2001; Experiment 1
present study) have found remote spatial memory to be impaired
Retention probes 2 and 3: quadrant analysis. On the second following hippocampal damage. However, the single study to use a
probe trial (administered approximately 15 min after the initial dry version of the water maze, where the rat must locate a specific
probe trial), all the control groups increased their percentage time point in space, found some evidence of spared remote memory
in the target quadrant (probe 2 performance for control groups; after hippocampal lesions (Kubie et al., 1999). Accordingly, in this
1-day ⫽ 80.4 ⫾ 3.8%; 8-week ⫽ 72.8 ⫾ 4.1%; 14-week ⫽ experiment we used a probe trial to assess recent and remote spatial
54.5 ⫾ 10.1%). This improvement might have been due to a memory for a land-based task following large hippocampal lesions.
reminder effect from probe trial 1 or to relearning (the probe trial We developed a land-based spatial memory task (Oasis maze) in
was reinforced). Interestingly, the 1-day and 8-week H-RF groups which a thirsty rat uses distal spatial cues to search an open field for
did not improve on probe 2 and remained at chance levels, but a specific location (Oasis) containing water (Fig. 1B). This task,
both the 8-week DH-RF group and the 14-week H-RF group which is conceptually similar to a previously described, food mo-
improved to above chance levels (8-week DH-RF ⫽ 53.9 ⫾ 8.8%; tivated task (Kesner et al., 1991), was designed to approximate the
14-week H-RF ⫽ 39.9 ⫾ 4.6%, t ⬎ 3.3, P ⬍ 0.05). Indeed, there spatial memory demands that are required by the water maze task
was no difference in performance on probe 2 between the 14-week (e.g., the open field of the Oasis maze was the same diameter as the
control group and the 14-week H-RF group (54.5 ⫾ 10.1% vs. water maze pool). We compared the performance of control ani-
____________________________________ HIPPOCAMPUS AND REMOTE SPATIAL MEMORY IN RATS 265

mals and animals with large hippocampal lesions made 1 day or At the beginning of each acquisition trial, the rats were placed at
9 weeks after training. For comparison, an additional group was one of four start points (North, South, East, and West) around the
included that received dorsal hippocampal lesions 9 weeks after edge of the board. Across the eight trials of each day (intertrial
training. interval was approximately 15 min), two trials were given from
each of the four start points, and the sequence was counterbalanced
Subjects within each group. A trial was terminated when the rat located the
We tested 106 experimentally naive, male Long-Evans rats water or after 5 min. If the rat failed to find the water within 5 min,
weighing 300 –350 g at the beginning of the experiment. Rats were it was guided to the correct well and allowed to drink. Once the
housed individually or in pairs and maintained on a 12:12-h light/ water was located, the rat was allowed to drink the water before
dark cycle. All rats had unlimited access to food (see water restric- being returned to the home cage. Following each training session,
tion below). the rats were allowed to drink freely for 1 h.
Following acquisition, rats in the spatial group were divided into
Apparatus equivalent surgery and control groups based on performance on
day 10, with the constraint that the animals in the surgery and
The Oasis maze was a circular, acrylic board (1.8 meters in control groups were equally likely to have been trained to find
diameter) that was painted flat white and raised 76 cm from the water in each quadrant.
floor by a table that allowed the board to be freely rotated on its
center axis. The surface of the board contained 426 evenly spaced Surgery and histology. The surgical procedures and coordi-
wells (2.5 cm in diameter, 1.3 cm in depth) in which small nates for the complete hippocampal lesion were the same as in
amounts of water (0.3 ml) could be hidden. Water was used as a Experiment 1. For rats given complete hippocampal lesions (H-
reward because, unlike food rewards, rats cannot locate the water RF), surgery was scheduled 1 day (n ⫽ 16; control group n ⫽ 16)
using olfactory cues (see results from RANDOM group below). or 9 weeks (n ⫽ 22; control group n ⫽ 32) after training. The
surgical procedures and coordinates for the dorsal hippocampal
Phase I: Pretraining lesions were the same as in Experiment 1. For rats given dorsal
Rats were water deprived by removing their water for 23 h/day hippocampal lesions (DH-RF), surgery was scheduled 9 weeks
for 3 days. Next, water was placed randomly in approximately 50% (n ⫽ 8) after training. The RANDOM group (n ⫽ 12) did not
of the wells (0.3 ml per well), and the rats were given 5 min to find undergo surgery. Following surgery, all rats were allowed 14 days
and drink as much water as they could on two occasions separated to recovery before retention probe testing. The neurohistological
by 30 min. The next day this procedure was repeated with only methods were the same as in Experiment 1.
25% of the wells containing water. On the final day, only five of
the wells contained water. Pretraining on this day ended when the
rats located and consumed water from all five baited wells.
Phase III: Retention probe
A 5-min probe trial was given 14 days after surgery. The trial
Phase II: Spatial training began by placing the rat at one of two start points around the edge
Following pretraining, rats were given eight trials a day for of the board. Start points were always in one of the two quadrants
10 days. On each trial, water was placed in a single well. For each adjacent to the quadrant that contained the training well location.
rat, the water was always located in the same position relative to the For example, for rats with training wells in the North, start points
testing room and distal spatial cues. Four different target well lo- for the probe trial were either West or East (counterbalanced
cations were used, one in the center of each quadrant, to control for within each group). No water was available during the probe trial.
any position biases the rats might have. The target well location Probe trials on the Oasis maze differed in two important ways
each rat was assigned to was balanced within each experimental from probe trials in the water maze. First, 5-min probe trials were
group. Local cues were made irrelevant (including odor trails) by used, rather than 1-min probe trials, to insure that rats explored a
rotating the entire apparatus a predetermined distance and direc- substantial portion of the Oasis maze. Second, in the Oasis maze
tion after each trial. However, the water was always placed in the rats had a strong tendency to explore the edges of the board and
same location relative to the testing room and distal spatial cues. also to return to the start point. For these reasons, chance perfor-
In addition to the groups trained to find water by using spatial mance cannot be calculated in a straightforward way. We esti-
cues, another group (RANDOM) was given an equivalent amount mated chance performance by assessing the performance of the
of training, except that the location of the water was varied ran- RANDOM group (because rats in this group could not acquire a
domly from trial to trial and could appear in any of the 426 wells. memory for a specific spatial location). Specifically, we compared
Thus, this group could develop optimal strategies for finding the the percentage of time that a trained rat spent within a 58-cm-
water efficiently, but memory of specific locations was irrelevant. diameter circle around the trained well with the percentage of time
The RANDOM group provided a measure of where rats with no that rats in the RANDOM group spent in this same region (target
spatial memory of a trained location will spend their search time locations for the RANDOM group were counterbalanced across
during probe trials. The probe trials for this group were adminis- rats). A 58-cm circle was the diameter of the largest circle that
tered 14 days after the end of training. could be completely contained within one quadrant of the maze.
266 CLARK ET AL.

Results

Histological findings
Figure 2B illustrates the extent of the largest and smallest lesion
in each group.

Hippocampal damage H-RF groups. All animals sustained

extensive bilateral damage to all the cell fields of the hippocampus,
including the dentate gyrus. The average percentage damage to the
hippocampus was 82.7% (range 70.5–93.0%). The pattern of
sparing was highly similar to the pattern reported in Experiment 1.

Other damage. Five animals sustained minor unilateral damage

to the entorhinal cortex that amounted to less than 10% of total
entorhinal volume. The entorhinal cortex was entirely spared in all FIGURE 4. Performance on the Oasis maze (Experiment 2). Per-
other animals. As in Experiment 1, in most cases there was some centage of time in the training circle during a retention probe trial
detectable damage to the cortical regions directly dorsal to the that was given 14 days after surgery. Control rats (CON) and rats with
dorsal hippocampus. Most frequently this damage was in the form complete hippocampal lesions (H-RF) or dorsal hippocampal lesions
(DH-RF) were tested for retention after a 1-day (CON, n ⴝ 16; H-RF,
of cortical thinning, but a few cases involved direct tissue damage. n ⴝ 16) or 9-week (CON, n ⴝ 24; H-RF, n ⴝ 22; DH-RF, n ⴝ 8)
Most animals sustained at least minor damage to the dorsal and training–surgery interval. None of the lesion groups performed better
ventral subiculum. One animal had bilateral damage to the medial than rats that were presented on each trial with random water loca-
and lateral habenula. No animal had any damage to the perirhinal tions (RANDOM, n ⴝ 12). Mean performance of the RANDOM
group is represented by the filled square. The 1-day and 9-week con-
cortex or the amygdala. trol groups performed better than the corresponding lesion groups
and better than the RANDOM group. Error bars ⴝ SEM.
Hippocampal damage DH-RF group. All animals sustained
extensive bilateral damage to all the cell fields of the dorsal hip-
pocampus, including the dentate gyrus. The average percentage Retention probe: circle analysis
damage to the hippocampus was 35.7% (range 33.5–39.5%). The
pattern of spared tissue was highly similar to the pattern described Figure 4 illustrates the percentage of time that each group spent
in the training circle during the 5-min retention probe trial that
in Experiment 1.
was given at different times after the completion of training. The
control groups exhibited forgetting across the intervals tested
Other damage. There was no substantial extrahippocampal
(1 day, 9.8 ⫾ 1.3% vs. 9 weeks, 6.4 ⫾ 0.9%; t[46] ⫽ 2.2, P ⬍
damage in this group, and no cortical thinning as noted in the
0.05.). Both scores were much better than the 3.1 ⫾ 0.7% score
H-RF groups.
obtained by the RANDOM group (t ⬎ 2.2, P ⬍ 0.05). By con-
trast, rats with complete hippocampal lesions (H-RF) or restricted
Behavioral Findings dorsal lesions (DH-RF) performed similarly to each other (t ⬍ 1.1,
P ⬎ 0.10) and no better than the RANDOM group (t ⬍ 0.6, P ⬎
Acquisition 0.10). The scores for the lesion groups were as follows: 1-day
The average distance that each group traveled to locate the H-RF ⫽ 4.4 ⫾ 2.9%; 9-week H-RF ⫽ 3.7 ⫾ 0.7%, 9-week
water across the first eight trials of training day 1 did not differ DH-RF ⫽ 3.1 ⫾ 0.8%.
between rats given spatial training and the rats trained with
random locations (trained groups ⫽ 1,035 ⫾ 75 cm; RAN- Discussion
DOM group ⫽ 1,349 ⫾ 75 cm, t[54] ⫽ 1.6, P ⬎ 0.1). By The control groups exhibited forgetting as time passed after
training day 2 however, the trained groups were already per- training and performed better than the RANDOM group at both
forming better than the RANDOM group (562 ⫾ 30 cm vs. retention intervals. Each lesion group performed more poorly than
1324 ⫾ 127 cm, t[54] ⫽ 8.5, P ⬎ 0.0001). By the tenth and the corresponding control group and no better than the RAN-
final day of training, the average distance traveled during all DOM group. Thus, the lesion groups failed to demonstrate reten-
eight trials was 198 ⫾ 8 cm for the trained groups and 640 ⫾ tion at either a short or long training–surgery interval.
110 cm for the RANDOM group. The improvement exhibited These findings are consistent with Experiment 1 and with pre-
by the RANDOM group reflects the development of more ef- vious studies that have used the water maze task (Bolhuis et al.,
ficient search strategies. The substantial improvement exhibited 1994; Mumby et al., 1999; Sutherland et al., 2001). However,
by the trained group reflects the learning of specific information these findings are at odds with the only previous study that used a
about the location of the water. dry land version of the water maze task (Kubie et al., 1999). This
____________________________________ HIPPOCAMPUS AND REMOTE SPATIAL MEMORY IN RATS 267

study reported spared remote spatial memory when hippocampal for 2 min, while the platform was not available. This procedure
lesions were made 14 weeks after training, but not 3 days after encouraged the rats to swim around the entire annulus.
training. In that study, rats had to search a large arena (1.83-m
diameter) to locate food buried in sawdust. One difference be- Phase II. Annular spatial training
tween the two studies is the length of the training–surgery interval
(9 weeks in our study vs. 14 weeks in the earlier study) and this Following pretraining, the curtains were removed to reveal the
difference might be an important factor. distal spatial cues. Rats were given eight trials each day for 10 days
(2 trials each day from the North, South, East, and West starting
points and with the sequence of starting points counterbalanced
EXPERIMENT 3: ANNULAR MAZE within groups). Rats were tested in squads of 8 (approximate in-
tertrial interval ⫽ 8 min). To escape the water, a rat was required to
In the annular water maze (Hollup et al., 2001), the rat is con- swim one complete lap around the pool. Once the lap was com-
fined to a circular corridor within a pool of water (Fig. 1C). The rat pleted, the escape platform was made available. After escaping, the
is trained to swim around the annulus, and an escape platform is rat remained on the platform for 30 s before being removed (or it
subsequently raised to just below the water level. A normal rat will was removed when it climbed onto the annulus walls and moved
learn the location of the hidden platform relative to distal land- away from the platform). The first and fifth trials of each day were
marks and will swim rapidly until it reaches the platform where it 60-s reinforced probe trials. During these trials, the platform was
can escape the water. Memory for the platform location can be initially in the lowered and inaccessible position. After 60 s elapsed
measured with probe trials while the platform is inaccessible. In and the rat had completed at least one lap, the platform was raised
that circumstance, memory is indicated when the rat decreases its to just below the water surface so that escape was possible.
swim speed (i.e., spends more time) in the platform location com- Following acquisition, rats were divided into equivalent surgery
pared to other zones of the annulus. The tendency to remain near and control groups based on performance on the first probe trial on
the platform location reveals that the rat has recognized the target day 10, with the constraint that each group had the same distribu-
location. Rats with hippocampal lesions made prior to training do tions of platform locations.
not acquire this tendency (Hollup et al., 2001). Using the annular
water maze, we compared performance of control animals and Surgery and Histology
animals with large hippocampal lesions made 9 weeks or 14 weeks
following training. The surgical procedures and coordinates for the complete hip-
pocampal lesion were the same as in Experiment 1. For this lesion,
surgery was scheduled 9 weeks (n ⫽ 7; control group n ⫽ 7) or
Subjects 14 weeks (n ⫽ 7; control group n ⫽ 7) after training. Rats were
We tested 28 experimentally naive, male Long-Evans rats allowed 14 days to recover before retention probe testing. The
weighing 300 –350 g at the beginning of the experiments. Rats neurohistological methods were the same as in Experiment 1.
were housed individually or in pairs and maintained on a 12:12-h
light/dark cycle. All rats had free access to food and water. Phase III: Retention probe testing
A probe trial was given 14 days after surgery. For probe trials, the
Apparatus annulus corridor was divided into 12 zones of 30° arcs (Fig 5A).
The annular maze consisted of a clear Plexiglas annulus (outer The platform was located in the center of one of the 30° zones
wall ⫽ 103-cm diameter, inner wall ⫽ 75-cm diameter) placed in (target zone). We measured the percentage of time that the rat
the center of the same pool of water used for the standard water spent in the target zone and the time spent in each of 11 other
maze task. The walls of the annulus extended 14 cm above the zones (chance performance ⫽ 8.3% per zone; Fig. 5A). The probe
water line and created a circular corridor that was 14 cm wide. The trial began by placing the rat immediately opposite the trained
distal spatial cues were the same as in Experiment 1. Two pneu- platform location and allowing the rat to swim for 2 min. The
matically controlled Atlantis platforms were placed on opposite timer began when the rat was placed in the pool and began to swim
sides of the pool within the annulus. When one platform was used away from the start point. During the probe trial, spatial memory
as a training location, the other remained in the lowered and inac- for the platform location was expressed as a reduction in swim
cessible position. Each rat received spatial training using one of the velocity when the rat was in the vicinity of the target zone. As a
two platform locations (counterbalanced within groups). result, the rat spent more time in this zone than in other zones.

Phase I: Pretraining Results

During pretraining, curtains surrounded the pool and blocked

Histological findings
the distal spatial cues. Each rat received four trials per day for
3 days. For each trial, the rat was placed at the North, South, East, Figure 2C illustrates the extent of the largest and smallest lesion
or West starting points and allowed to swim around the annulus in each group.
268 CLARK ET AL.

FIGURE 5. Performance on the annular water maze (Experi- training zone replotted as a function of training–surgery interval. For
ment 3). A: For scoring, twelve 30° arc zones were constructed around rats with complete hippocampal lesions (H-RF), surgery occurred
the training (platform location) zone. Chance performance is 8.3% 9 weeks or 14 weeks after training (n ⴝ 7 for all groups). None of the
(100% divided by 12 zones). B: Percentage time spent by each group lesion groups performed above chance (8.3%). Both the 9-week CON
in the 12 zones during the retention probe trial that was given 14 days and the 14-week CON groups performed better than the correspond-
after surgery. Memory of the platform location is indicated as in- ing lesion groups. Error bars ⴝ SEM.
creased percentage time in the training zone. C: Percentage time in the

Hippocampal damage H-RF groups. All animals sustained group spent in the target zone as a function of training–surgery
extensive bilateral damage to all the cell fields of the hippocampus, interval. The control groups spent 22.6 ⫾ 4.3% and 14.4 ⫾ 3.0%
including the dentate gyrus. The average percentage damage to the of the time in the target zone in the 9-week and 14-week condi-
hippocampus was 87.4% (range 76.4 –91.9%). The pattern of tions, respectively. The 9-week control group performed above
sparing was highly similar to the pattern reported in Experiment 1. chance (t[6] ⫽ 3.3, P ⬍ 0.05) and better than the 9-week H-RF
group (8.3 ⫾ 0.6%; t[12] ⫽ 3.2, P ⬍ 0.01). The 14-week control
Other damage. One animal sustained minor unilateral damage group performed marginally above chance (t[6] ⫽ 2.0, P ⫽ 0.09)
to the entorhinal cortex that amounted to less than 5% of total and better than the 14-week H-RF group (7.7 ⫾ 0.7%; t[12] ⫽
entorhinal volume. The entorhinal cortex was entirely spared in all 2.2, P ⫽ 0.05). The two groups with hippocampal lesions per-
other animals. As in Experiments 1 and 2, most cases showed some formed similarly to each other and no better than chance (t ⬍ 0.13,
detectable damage to the cortical regions directly dorsal to the P ⬎ 0.10).
dorsal hippocampus. Most frequently this damage was in the form
of cortical thinning. Most animals sustained at least minor damage Discussion
to the dorsal and ventral subiculum. In two cases, there was some
The control groups exhibited forgetting as time passed after
minor unilateral damage to the lateral dorsal nucleus of the thala-
learning. Each lesion group performed more poorly than the cor-
mus. No animal had any damage to the perirhinal cortex or the
responding control group and no better than chance. Thus, the
amygdala.
lesion group exhibited no evidence of retention at either training–
surgery interval (Fig. 5C). It was previously reported that hip-
Behavioral findings
pocampal lesions impair anterograde memory on this task (Hollup
Acquisition. On the first probe trial (trial 1) of spatial training, et al., 2001). The present work extends these findings and shows
rats spent 9.8 ⫾ 1.0% of the time in the target zone. This score was that hippocampal lesions also impair retrograde memory.
not above chance (chance ⫽ 8.3%, t[27] ⫽ 1.5, P ⬎ 0.10). On the
first probe trial on the tenth and final day of training, rats spent
19.7 ⫾ 1.9% of the time in the target zone. This score was well GENERAL DISCUSSION
above chance (chance ⫽ 8.3%, t[27] ⫽ 6.0, P ⬍ 0.0001) and
better than performance on the first probe trial on the first day of The findings were clear and consistent across three experiments
training (t[27] ⫽ 5.1, P ⬍ 0.0001). that examined recent and remote spatial memory following dam-
age to the hippocampus. The results in the standard water maze
Retention probe. Figure 5B shows the percentage of time that (Experiment 1) revealed no evidence of spared spatial memory
each group spent in each zone during the 2-min retention probe with training–surgery intervals of 1 day, 8 weeks or 14 weeks.
trial. Notice the elevated percentage of time the control rats spent Similarly, the results in the Oasis maze (Experiment 2) revealed no
in the target zone. Figure 5C shows the percentage of time each evidence of spared spatial memory with training–surgery intervals
____________________________________ HIPPOCAMPUS AND REMOTE SPATIAL MEMORY IN RATS 269

of 1 day or 9 weeks. Finally, in the annular maze (Experiment 3), derstand the present results, and there are also other findings that
there was no evidence of spared spatial memory with training– must be considered in any discussion of remote spatial memory
surgery intervals of 9 weeks or 14 weeks. In summary, there was no and hippocampal function.
tendency for the lesion groups, in any of the experiments, to per- One important finding is that in humans, remote spatial mem-
form well at the longer training–surgery intervals. Further, rela- ory was spared even following large medial temporal lobe lesions.
tively small, dorsal hippocampal lesions impaired performance to Thus, patient E.P. was able to recall the spatial layout of the region
the same extent as complete lesions. where he grew up and from which he moved away as a young adult
Two points merit discussion. First, our lesion method (radiofre- more than 50 years earlier. He could mentally navigate, construct
quency) damaged both cell bodies and fibers. We were concerned novel routes, and point correctly to landmarks while imagining
that excitotoxic lesions might damage structures, and possibly im- himself at various locations. Yet E.P. has no knowledge of the
pair remote memories, outside of the hippocampus as the result of neighborhood where he has lived since 1993, the year after he
the sustained hyperactivity of hippocampal neurons following the became amnesic (Teng and Squire, 1999). In another study, pa-
injection of a fiber-sparing excitotoxin (e.g., Anagnostaras et al., tient K.C., who has bilateral hippocampal damage, as well as other
2001). Further, because most fiber bundles within the hippocam- significant damage, can retrieve remotely formed spatial memories
pus have hippocampal neurons as their origin or target (Amaral of major landmarks, construct routes, and estimate distances and
and Witter, 1995), most of these fiber bundles would have been directions in the neighborhood in which he grew up and has lived
compromised by either a radiofrequency lesion or an excitotoxic for approximately 40 years (Rosenbaum et al., 2000). These find-
lesion. Nonetheless, the possibility remains that our results might ings show that the human medial temporal lobe is needed to ac-
have differed if fiber-sparing lesions had been used. Second, the quire new spatial knowledge (Teng and Squire, 1999), but that it is
effects of dorsal hippocampal lesions (as opposed to complete le- not the repository of remotely acquired spatial maps (Teng and
sions) were studied only at an 8-week (water maze, Experiment 1) Squire, 1999; Rosenbaum et al., 2000). One difference between
and 9-week (oasis maze, Experiment 2) training–surgery interval. these studies in humans and our studies in rats is that spatial learn-
It is possible that had the effects of dorsal hippocampal lesions been ing in the rats occurred during a limited period of time when the
studied in all three tasks and also at shorter or longer training– animals were adults, whereas the spatial learning studied in the
surgery intervals, some sparing of remote memory might have been patients was acquired beginning at an early age and the learning
observed. Nonetheless, the results following complete hippocam- continued over many years. Perhaps spatial memories must be very
pal lesions as well as the results following dorsal hippocampal le- well learned over a long period of time, or perhaps they must be
sions were unambiguous and provided no evidence of spared re- acquired early in life (or both) if they are to be spared following
mote spatial memory. hippocampal damage (for a thoughtful discussion of these possi-
These findings can be contrasted with the pattern of results that bilities, and others, see Rosenbaum et al. (2001).
have typically been observed following hippocampal damage on Although there was little or no evidence of spared remote spatial
tests of nonspatial memory, such as trace eyeblink conditioning memory in the present study, other studies have clearly demon-
(Kim et al., 1995; Takehara et al., 2002, 2003). In these cases, strated spared remote memory following hippocampal lesions in
hippocampal lesions made soon after training impaired perfor- rats in tasks that appear to require some form of spatial memory.
mance, and lesions made one month after training had no effect. A For example, spared remote (and impaired recent ) memory has
number of other similar findings have been reported (for review, consistently been reported when spatial information must be used
see Squire et al., 2004). For example, in the odor-odor association to guide a simple two-choice (Cho et al., 1993, 1995) or three-
task known as the social transmission of food preference (Galef and choice (Ramos, 1998) spatial discrimination between arms of a
Wigmore, 1983), temporal gradients of retrograde amnesia cover- maze. Additionally, in an 8-arm spatial discrimination task in
ing 1–5 days have been observed following lesions of the dorsal mice, brain imaging ([14C]2-deoxyglucose uptake) revealed greater
hippocampus (Winocur, 1990) and following larger lesions that hippocampal activation during a retention test 5 days after learning
damaged the dorsal and ventral hippocampus (Winocur et al., (recent spatial memory) than during a retention test 25 days after
2001). When the damage was even more extensive, including dor- learning (remote spatial memory) (Bontempi et al., 1999). Finally,
sal and ventral hippocampus and substantial portions of the dorsal infusion of lidocaine into dorsal hippocampus 1 day after training
and ventral subiculum, a 1–30-day gradient of retrograde amnesia impaired memory for the trained arm of a 5-arm maze, but infu-
was observed (Clark et al., 2002). sion after 30 days had no effect (Maviel et al., 2004). These results
On first examination, the present findings appear broadly con- are not consistent with the view that spatial memory is perma-
sistent with the view, which grew out of the discovery of hip- nently stored in the hippocampus.
pocampal place cells in the rat, that the hippocampus is essential Contextual fear conditioning also involves spatial memory, be-
for forming and storing spatial maps (O’Keefe and Nadel, 1978). cause the rat explores a novel environment and encodes numerous
By this view, temporally graded retrograde amnesia should not stimulus features, including visuospatial information, which must
occur for spatial memory because spatial maps (i.e., spatial mem- be integrated in order to represent a context (Fanselow, 2000).
ories) thought to be stored in the hippocampus are destroyed when Temporal gradients of retrograde amnesia (i.e., spared remote
the hippocampus is damaged. Accordingly, a hippocampal lesion memory and impaired recent memory) have consistently been ob-
should impair spatial memory, no matter how long after learning served following hippocampal damage in the case of classically
the lesion is made. However, there are other possible ways to un- conditioned fear to a context (Kim and Fanselow, 1992; Maren et
270 CLARK ET AL.

al., 1999; Anagnostaras et al., 1999). In support of these studies, which can be impaired after fornix lesions, and “knowing
findings with ␣-CaMKII⫹/⫺ mutant mice and activity-dependent where,” which can be spared (e.g., Whishaw et al., 1995). Rats
genes suggest that both contextual fear conditioning and spatial with fornix lesions were impaired in learning how to locate a
learning in the water maze become progressively more dependent hidden platform as opposed to learning where a platform is
on the neocortex as time passes after learning (Frankland et al., located.
2001, 2004). Rats with hippocampal damage also exhibited marked impair-
One feature that may distinguish tasks where remote spatial ments in path integration (Whishaw and Maaswinkel, 1998;
memory is spared following hippocampal lesions from tasks where Maaswinkel et al., 1999). In path integration (also referred to as
remote spatial memory is abolished involves how animals use spa- dead-reckoning; Darwin 1873), an animal continually updates
tial information. Interestingly, only in those tasks where the ex- and computes its position in space relative to some starting point
pression of memory requires the animal to move to a specific spatial by integrating the body movement cues generated by physical lo-
location has remote spatial memory been impaired following hip-
comotion (for review, see Etienne and Jeffery, 2004). In addition,
pocampal lesions (see Kubie et al., 1999, for the single exception).
many hippocampal neurons exhibit physiological properties that
In tasks where animals must use spatial information to discrimi-
would be useful for navigation. Thus, in addition to place cells, the
nate between two or three maze arms, rather than to find a specific
hippocampus possesses head direction cells, and the firing proper-
location, remote memory is spared (Cho et al., 1993, 1995;
Ramos, 1998; Maviel et al., 2004). Here we consider two possible ties of these cells are influenced by body movements (for review, see
reasons why remote spatial memory is impaired by hippocampal Redish, 2001; Etienne and Jeffery, 2004). These observations pro-
lesions in the first case, but not in the second case. vide additional reasons for supposing that hippocampal lesions
First, remote spatial memory might survive hippocampal le- might impair performance whenever a task requires navigation.
sions, but be reduced in quality or quantity. By this account, the At first glance, it might appear that the findings of Experiment 3
surviving spatial information is sufficient to guide a spatial discrim- count against this proposal that hippocampal lesions impair per-
ination between maze arms, or to reactivate conditioned fear of a formance by impairing the new learning that is needed to navigate
context (Kim and Fanselow, 1992; Maren et al., 1999; Anagnost- and express a remote spatial memory. In Experiment 3, remote
aras et al., 1999), but not sufficient to guide navigation to a specific spatial memory was impaired even though the annular maze would
point in space. This idea is consistent with the view that the hip- seem to minimize the need for navigation. Animals must simply
pocampus stores the rich spatial details (maps) that are critical for swim until they encounter (and recognize) the target location. Yet,
guiding an animal to a specific location (O’Keefe and Nadel, it is unknown how animals actually accomplish this task. It is true
1978). that the annular maze task is less sensitive to hippocampal disrup-
Yet, another possibility is that remote spatial memory fully sur- tion than the standard water maze task (Brun et al., 2002), but this
vives hippocampal damage, but that the lesion impairs the expres- finding is consistent with the possibility that normal animals in this
sion of spatial memory (i.e., the ability to perform the task). By this task demonstrate their spatial memory by continually updating
view, the navigational demands of expressing spatial memory re- their position in space relative to distal spatial cues, just as in the
quire an intact hippocampus, perhaps because navigating to a spe- standard water maze. By this view, animals with hippocampal le-
cific point requires new learning. In other words, the animal must sions are unable to update their position in space, do not know
be able to update (encode) its position in space continually in order where they are, and thus do not recognize the target location when
for a specific spatial memory to be expressed (Knowlton and they approach it.
Fanselow, 1998). In the same sense, an amnesic patient would not
It will be difficult to test the idea that hippocampal lesions
be expected to be able to express an otherwise intact spatial mem-
impair performance, i.e., the expression of memory, in animals
ory, if the performance test required executing a number of turns
with permanent hippocampal lesions. However, in other para-
and moving along several routes. In this case, the memory demands
digms, reversible lesions have been successful in distinguishing
of the performance test would exceed what can be maintained in
between impaired memory and the impaired expression of
immediate memory.
In contrast, the ability to accomplish new learning would not be memory (Clark et al., 1992; Clark and Lavond, 1993; Krupa et
required for spatial discrimination tasks where the animal needs al., 1993). Reversible lesions afford the important advantage
only to choose the correct arm of a maze. Once the correct arm is that retention can be tested after the lesion has been reversed
chosen, the animal needs only to proceed along the arm to obtain and during a time when the hippocampus is functional. Some
food. Similarly in the case of contextual fear conditioning, memory progress has been made using reversible lesions (e.g., with an
is measured by the amount of freezing (no locomotion is required), AMPA antagonist) in studies of spatial memory (Riedel et al.,
and the performance test does not require the acquisition of new 1999; Maviel et al., 2004).
information. In the same sense, amnesic patients do not need to
acquire new information in order to answer questions about their Acknowledgments
remote spatial memory.
These considerations raise the possibility that hippocampal The authors thank Laura Entwistle, Marie-Claire Doré, Gene
lesions impair task performance rather than memory itself. This Delay, Natalie Shanks, Daniel Guadarrama, Darwin-Dean
idea is reminiscent of the distinction between “getting there,” Castillo, and Stuart Zola for assistance.
____________________________________ HIPPOCAMPUS AND REMOTE SPATIAL MEMORY IN RATS 271

Kapur N, Brooks DJ. 1999. Temporally-specific retrograde amnesia in

REFERENCES two cases of discrete bilateral hippocampal pathology. Hippocampus
9:247–254.
Kesner RP, Farnsworth G, Kametani H. 1991. Role of parietal cortex and
Amaral DG, Witter MP. 1995. Hippocampal formation. In: Paxinos G, hippocampus in representing spatial information. Cereb Cortex
editor. The rat nervous system. 2nd ed. San Diego, CA: Academic 1:367–373.
Press. p 443– 493. Kim JJ, Fanselow MS. 1992. Modality-specific retrograde amnesia of fear.
Anagnostaras SG, Maren S, Fanselow MS. 1999. Temporally graded ret- Science 256:675– 677.
rograde amnesia of contextual fear after hippocampal damage in rats: Kim JJ, Clark RE, Thompson RF. 1995. Hippocampectomy impairs the
within-subjects examination. J Neurosci 19:1106 –1114. memory of recently, but not remotely, acquired trace eyeblink condi-
Anagnostaras SG, Gale GD, Fanselow MS. 2001. Hippocampus and con- tioned responses. Behav Neurosci 109:195–203.
textual fear conditioning: recent controversies and advances. Hip- Knowlton BJ, Fanselow MS. 1998. The hippocampus, consolidation and
pocampus 11:8 –17. on-line memory. Curr Opin Neurobiol 8:293–296.
Bolhuis J, Stewart CA, Forrest EM. 1994. Retrograde amnesia and mem- Krupa DJ, Thompson JK, Thompson RF. 1993. Localization of a mem-
ory reactivation in rats with ibotenate lesions to the hippocampus or ory trace in the mammalian brain. Science 260:989 –991.
subiculum. Q J Exp Psychol B 47:129 –150. Kubie JL, Sutherland RJ, Muller RU. 1999. Hippocampal lesions produce
Bontempi B, Laurent-Demir C, Destrade C, Jaffard R. 1999. Time- a temporally graded retrograde amnesia on a dry version of the Morris
dependent reorganization of brain circuitry underlying long-term swimming task. Psychobiology 27:313–330.
memory storage. Nature 400:671– 675. Maaswinkel H, Jarrard LE, Whishaw IQ. 1999. Hippocampectomized
Broadbent NJ, Squire LR, Clark RE. 2004. Spatial memory, recognition rats are impaired in homing by path integration. Hippocampus 9:553–
memory, and the hippocampus. Proc Natl Acad Sci USA 101:14515– 561.
14520. Manns JR, Hopkins RO, Reed JM, Kitchener EG, Squire LR. 2003.
Brun VH, Otnass MK, Molden S, Steffenach HA, Witter MP, Moser Recognition memory and the human hippocampus. Neuron 37:1–20.
MB, Moser EI. 2002. Place cells and place recognition maintained by Maren S, Aharonov G, Fanselow MS. 1999. Neurotoxic lesions of the
direct entorhinal-hippocampal circuitry. Science 296:2243–2246. dorsal hippocampus and Pavlovian fear conditioning in rats. Behav
Cho YH, Beracochea D, Jaffard R. 1993. Extended temporal gradient for Brain Res 110:436 – 442.
the retrograde and anterograde amnesia produced by ibotenate ento- Maviel T, Durkin T, Menzaghi F, Bontempi B. 2004. Sites of neocortical
rhinal cortex lesions in mice. J Neurosci 13:1759 –1766.
reorganization critical for remote spatial memory. Science 305:96 –99.
Cho YH, Kesner RP, Brodale S. 1995. Retrograde and anterograde am-
Morris RGM. 1984. Developments of a water-maze procedure for study-
nesia for spatial discrimination in rats: role of hippocampus, entorhinal
ing spatial learning in the rat. J Neurosci Methods 11:47– 60.
cortex, and parietal cortex. Psychobiol 23:185–194.
Moser E, Moser M-B, Andersen P. 1993. Spatial learning impairment
Clark RE, Lavond DG. 1993. Reversible lesions of the red nucleus during
parallels the magnitude of dorsal hippocampal lesions, but is hardly
acquisition and retention of a classically conditioned behavior in rab-
present following ventral lesions. J Neurosci 13:3916 –3925.
bits. Behav Neurosci 107:264 –270.
Moser M-B, Moser EI, Forrest E, Andersen P, Morris RGM. 1995. Spatial
Clark RE, Zhang AA, Lavond DG. 1992. Reversible lesions of the cere-
learning with a minislab in the dorsal hippocampus. Proc Natl Acad
bellar interpositus nucleus during acquisition and retention of a clas-
sically conditioned behavior. Behav Neurosci 106:879 – 888. Sci USA 92:9697–701.
Clark RE, Zola SM, Squire LR. 2000. Impaired recognition memory in Mumby DG, Astur RS, Weisend MP, Sutherland RJ. 1999. Retrograde
rats after damage to the hippocampus. J Neurosci 20:8853– 8860. amnesia and selective damage to the hippocampal formation: memory
Clark RE, Broadbent NJ, Zola-Morgan S, Squire LR. 2002. Anterograde for places and object discriminations. Behav Brain Res 106:97–107.
amnesia and temporally-graded amnesia for a nonspatial memory task O’Keefe J, Nadel L. 1978. The hippocampus as a cognitive map. London:
following lesions of hippocampus and subiculum. J Neurosci Oxford University Press.
22:4663– 4669. Ramos J. 1998. Retrograde amnesia for spatial information: dissociation
Darwin C. 1873. On the origin of certain instincts. Nature 7:417– 418. between intra and extramaze cues following hippocampus lesions in
de Hoz L, Knox J, Morris RGM. 2003. Longitudinal axis of the hip- rats. Eur J Neurosci 10:3295–3301.
pocampus: both septal and temporal poles of the hippocampus sup- Redish AD. 2001. The hippocampal debate: are we asking the right ques-
port water maze spatial learning depending on the training protocol. tions? Behav Brain Res 127:81–98.
Hippocampus 13:587– 603. Ribot T. 1881. Les maladies de la memoire [English translation: Diseases
Etienne AS, Jeffery KJ. 2004. Path integration in mammals. Hippocam- of memory]. New York: Appleton-Century-Crofts.
pus 14:180 –192. Riedel G, Micheau J, Lam AG, Roloff E, Martin SJ, Bridge H, de Hoz L,
Fanselow MS. 2000. Contextual fear, gestalt memories, and the hip- Poeschel B, McCulloch J, Morris RGM. 1999. Reversible neural in-
pocampus. Behav Brain Res 110:73– 81. activation reveals hippocampal participation in several memory pro-
Frankland PW, O’Brien C, Ohno M, Kirkwood A, Silva AJ. 2001. Alpha- cesses. Nat Neurosci 2:898 –905.
CaMKII-dependent plasticity in the cortex is required for permanent Rosenbaum RS, Prisela, S, Kohler S, Black SE, Gao F, Nadel L, Mosco-
memory. Nature 411:309 –313. vitch M. 2000. Remote spatial memory in an amnesic person with
Frankland PW, Bontempi B, Talton LE, Kaczmarek L, Silva AJ. 2004. extensive bilateral hippocampal lesions. Nat Neurosci 3:1044 –1048.
The involvement of the anterior cingulate cortex in remote contextual Rosenbaum RS, Winocur G, Moscovitch M. 2001. New views on old
fear memory. Science 304:881– 883. memories: re-evaluating the role of the hippocampal complex. Behav
Galef BG, Wigmore SR. 1983. Transfer of information concerning dis- Brain Res 127:183–197.
tant foods: a laboratory investigation of the “information-centre” hy- Spooner RIW, Thomson A, Hall J, Morris RGM, Salter SH. 1994. The
pothesis. Anim Behav 31:748 –758. Atlantis platform: a new design and further developments of Bureso-
Hollup SA, Kjelstrup KG, Hoff J, Moser M-B, Moser E. 2001. Impaired va’s on-demand platform for the water maze. Learn Mem 3:203–211.
recognition of the goal location during spatial navigation in rats with Squire LR, Alvarez P. 1995. Retrograde amnesia and memory consolida-
hippocampal lesions. J Neurosci 12:4505– 4513. tion: a neurobiological perspective. Curr Opin Neurobiol 5:169 –177.
Kapur N. 1993. Focal retrograde amnesia in neurological disease: a critical Squire LR, Clark RE, Knowlton BJ. 2001. Retrograde amnesia. Hip-
review. Cortex 29:217–234. pocampus 11:50 –55.
272 CLARK ET AL.

Squire LR, Clark RE, Bayley PJ. 2005. Medial temporal lobe function and Teng E, Squire LR. 1999. Memory for places learned long ago is intact
memory. In: Gazzinaga M, editor. The cognitive neurosciences. 3rd after hippocampal damage. Nature 400:675– 677.
ed. Cambridge: MIT Press p 691–708. Whishaw IQ, Maaswinkel H. 1998. Rats with fimbria-fornix lesions are
Sutherland RJ, Weisend MP, Mumby D, Astur RS, Hanlon FM, Ko- impaired in path integration: a role for the hippocampus in “sense of
erner A, Thomas MJ, Wu Y, Moses SN, Cole C, Hamilton DA, direction.” J Neurosci 18:3050 –3058.
Hoesing JM. 2001. Retrograde amnesia after hippocampal damage: Whishaw IQ, Cassel JC, Jarrad LE. 1995. Rats with fimbria-fornix lesions
recent vs. remote memories in three tasks. Hippocampus 11:27– display a place response in a swimming pool: a dissociation between
42. getting there and knowing where. J Neurosci 15:5779 –5788.
Takehara K, Kawahara S, Takatsuki K, Kirino Y. 2002. Time-limited role Winocur G. 1990. Anterograde and retrograde amnesia in rats with dorsal
of the hippocampus in the memory for trace eyeblink conditioning in hippocampal or dorsomedial thalamic lesions. Behav Brain Res 38:
mice. Brain Res 951:183–190. 145–154.
Takehara K, Kawahara S, Kirino Y. 2003. Time-dependent reorganization Winocur G, McDonald RM, Moscovitch M. 2001. Anterograde and
of the brain components underlying memory retention in trace eye- retrograde amnesia in rats with large hippocampal lesions. Hippocam-
blink conditioning. J Neurosci 23:9897–9905. pus 11:18 –26.