The Journal of Neuroscience, December 1989, 9(12): 4355-4370 Lesions of Perirhinal and Parahippocampal Cortex That Spare the Amygdala and Hippocampal Formation Produce Severe Memory Impairment Stuart Zola-Morgan,’ Larry Ft. Squire,’ David G. Amaral,2 and Wendy A. Suzuki2J Veterans Administration Medical Center, San Diego, California, 92161, and Department of Psychiatry, University of California, San Diego, La Jolla, California 92093, The Salk Institute, San Diego, California 92136, and 3Group in Neurosciences, University of California, San Diego, La Jolla, California 92093 In monkeys, bilateral damage to the medial temporal region Moss, 1984). (In this notation, H refers to the hippocampus, A produces severe memory impairment. This lesion, which in- to the amygdala, and the plus superscript (+) to the cortical cludes the hippocampal formation, amygdala, and adjacent tissue adjacent to each structure.) This lesion appears to con- cortex, including the parahippocampal gyrus (the H+A+ le- stitute an animal model of medial temporal lobe amnesia like sion), appears to constitute an animal model of human me- that exhibited by the well-studied patient H.M. (Scoville and dial temporal lobe amnesia. Reexamination of histological Milner, 1957). material from previously studied monkeys with H+A+ lesions The H+A+ lesion produces greater memory impairment than indicated that the perirhinal cortex had also sustained sig- a lesion limited to the hippocampal formation and parahip- nificant damage. Furthermore, recent neuroanatomical stud- pocampal cortex-the H+ lesion (Mishkin, 1978; Mahut et al., ies show that the perirhinal cortex and the closely associated 1982; Zola-Morgan and Squire, 1985, 1986; Zola-Morgan et al., parahippocampal cortex provide the major source of cortical 1989a). One possibility to explain these findings is that the more input to the hippocampal formation. Based on these 2 find- severe deficit occurs because the amygdala is damaged in the ings, we evaluated the severity of memory impairment in a larger H+A+ lesion but not in the more selective H+ lesion (Mish- group of monkeys that received bilateral lesions limited to kin, 1978; Murray and Mishkin, 1984, 1986; Saunders et al., the perirhinal cortex and parahippocampal gyrus (the PRPH 1984; Bachevalier et al., 1985). However, we recently found that lesion). The performance of the PRPH group was compared stereotaxic bilateral lesions of the amygdala, which spared ad- with that of monkeys with H+A+ lesions, who had been stud- jacent cortex, did not impair performance on 4 different memory ied previously, and with a group of normal monkeys. Mon- tasks and did not exacerbate the memory impairment associated keys with PRPH lesions were severely impaired on 3 am- with hippocampal formation (H+) lesions alone (Zola-Morgan nesia-sensitive tasks: delayed nonmatching to sample, object et al., 1989b). retention, and d-pair concurrent discrimination. On pattern A second possibility is that the H+A+ lesion produces more discrimination, a task analogous to ones that amnesic pa- severe memory impairment than the H+ lesion because of dam- tients perform well, monkeys in the PRPH group performed age to perirhinal cortex rather than because of damage to the normally. Overall, monkeys with PRPH lesions were as im- amygdala. Reexamination of the histological material from one paired or more impaired than the comparison group of mon- of the studies involving the H+A+ lesion @la-Morgan et al., keys with H+A+ lesions. These and other recent findings 1982) indicated that the perirhinal cortex sustained substantial (Zola-Morgan et al., 1989b) suggest that the severe memory damage (see the histological findings in Results). Furthermore, impairment in monkeys and humans associated with bilat- recent neuroanatomical studies show that the perirhinal cortex eral medial temporal lesions results from damage to the (areas 35 and 36) and the closely associated parahippocampal hippocampal formation and adjacent, anatomically related cortex (areas TH and TF) provide the major source of cortical cortex, not from conjoint hippocampus-amygdala damage. input to the entorhinal cortex (Insausti et al., 1987). These 2 cortical regions thus provide the principal route by which in- Bilateral damage to the hippocampal formation, amygdala, and formation is exchanged between the neocortex and the hippo- adjacent cortex, including the parahippocampal gyrus (the H+A+ campal formation. lesion), produces severe memory impairment in monkeys In the present study, we evaluated the severity of memory (Mishkin, 1982; Squire and Zola-Morgan, 1983; Mahut and impairment in a group of monkeys that received conjoint bi- lateral lesions limited to the perirhinal cortex and the parahip- pocampal gyrus (the PRPH lesion). The performance of this Received Mar. 8, 1989; revised June 9, 1989; accepted June 16, 1989. group was compared with the performance of monkeys with the This work was supported by the Medical Research Service of the Veterans H+A+ lesion that had been studied previously (Zola-Morgan et Administration, NIH grants 19063, NS 16980, and MH 4 1479, the Office ofNaval Research, and the M&night Foundation. We thank Melodee Strukelj, Jasmine al., 1982; Zola-Morgan and Squire, 1984, 1985) and with a Allen, Michael Mesches, Amy Lockwood, and Melissa Hsu for behavioral testing, group of normal monkeys. Performance was tested on the de- and Cecelia Leclair and Janet Weber for histoloaical assistance. Correspondence should be addressed to Stua> Zola-Morgan, Department of layed nonmatching to sample task, on 2 other tasks sensitive to Psychiatry M-003, U.C.S.D. School of Medicine, La Jolla, CA 92093. human amnesia (delayed retention of object discriminations and Copyright 0 1989 Society for Neuroscience 0270-6474/89/124355-16$02.00/O 8-pair concurrent discrimination learning; Squire et al., 1988), 4356 Zola-Morgan et al. l Cortex Lesions That Impair Memory and on a task analogous to ones that amnesic patients perform Wisconsin General Test Apparatus (Harlow and Bromer, 1938). Four well (pattern-discrimination learning; Zola-Morgan and Squire, different tasks-i.e., trial-unique delayed nonmatching to sample, pat- tern discrimination, delayed retention of object discriminations, and 1984). concurrent discrimination learning-were administered to all monkeys Materials and Methods in the order listed under Behavioral findings. These tasks are described in detail in Zola-Morgan et al. (1989a). Subjects The findings from 14 cynomolgus monkeys (Mucucu fascicularis)will be presented. All monkeys weighed between 3.0 and 4.8 kg at the be- Retrograde tracer studies ginning of behavioral testing and were estimated to be 3-5 years old, In order to determine the extent to which the intended PRPH lesion i.e., young adults (Hartley et al., 1984; Szabo and Cowan, 1984). had deafferented the hippocampal formation (particularly the entorhinal The 14 monkeys belonged to 3 experimental groups. Four male mon- cortex) from its cortical inputs, in 2 of the 4 PRPH animals the ento- keys received bilateral lesions of the PRPH. The intent was to spare the rhinal cortex was injected with a retrograde tracer. The contralateral amygdaloid complex and the hippocampal formation (dentate gyrus, amygdala was injected with a second retrograde tracer to insure that hippocampus proper, subicular complex, and entorhinal cortex). The 4 significant loss of corticoamygdaloid connections had not occurred due monkeys in the H+A+ group (2 males, 2 females) received bilateral to inadvertent damage to fibers traveling within or close to the lesioned lesions with the intent to remove the amygdala and all subdivisions of area. Following surgical procedures similar to those described above, the hippocampal formation, as well as much of the parahippocampal animals PRPHl and PRPH3 were placed in a Kopf stereotaxic appa- cortex. Behavioral data for this group have been presented as part of 3 ratus. Using coordinates derived from the atlas of Szabo and Cowan previous reports (Zola-Morgan et al., 1982; Zola-Morgan and Squire, (1984) and guided by electrophysiological recording along the trajectory 1984. 1985) and are included here for purposes of comparison. Finally, of the intended injection site, injections of retrograde tracers were placed 6 monkeys (5 males, 1 female) were unoperated and comprised a normal in the entorhinal cortex of one side and in the amygdaloid complex on control group (IV). Three of these monkeys (N4, N5, N6) were tested at the other side. In PRPH 1, 500 nl of 2.0% Diamidino yellow (DY) was the same time as the monkeys in the PRPH group. The other 3 normal placed into the entorhinal cortex on the left side, and 500 nl of 3.0%. monkeys (N 1, N2, N3) were-tested at the same time as the H+A+ group. Fast blue (FB) was aimed at the lateral nucleus of the amygdala on the Behavioral data for Nl , N2, and N3 have also been reported previously right side. The animal was allowed to survive 2 weeks after the injec- (Zola-Morgan and Squire, 1984, 1985; Zola-Morgan et al., 1989a, b). tions. In PRPH3, the amygdaloid injection was similar (though on the left side), but the entorhinal injection on the right side consisted of 100 nl of a 2% solution of WGA-HRP. The survival time after the DY Surgery injection was 14 d and, after the WGA-HRP injection, 2 d (the 2 tracers PRPH group. At the time of surgery, animals were preanesthetized with were placed in separate surgeries to accommodate the different survival ketamine hvdrochloride (12-15 ma& i.m.) and then brought to a times). surgical plane of anesthesia with in&~enous administration of Nem- butal (25-30 mg&). Heart rate and body temperature were monitored continuously, and additional anesthetic was administered when nec- Histological processing essary. The animals were killed by transcardial perfusion of fixatives. Following The monkey’s head was positioned in a specially designed headholder deep anesthetization and loss of the comeal reflex, the animals were that permitted unobstructed access to the temporal portion of the skull.