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Brief ganglioside treatment produces delayed enhancement of functional recovery after medial septal lesions
396
Brain Research. 495 (19~t/) 39{~-40(I Elsevier
BRES 23673
Brief ganglioside treatment produces delayed enhancement of functional recovery after medial septal lesions A l e x P o p l a w s k y I a n d R o b e r t L. Isaacson 2 1Department of Psychology, Bloomsburg University of Pennsylvania, Bloomsburg, PA 17815 (U. S. A. ) and 2Department of Psychology, State University of New York at Binghamton, Binghamton, NY 13901 (U.S.A.) (Accepted 2 May 1989) Key words: Medial septal lesion; Ganglioside; Recovery from brain damage; Body weight; Activity; Stereotypy
The effects of a 5-day ganglioside (GMI) treatment (30 mg/kg) on body weight and water intake subsequent to medial septal lesions were evaluated for 44 consecutive days. In addition, activity, rearing, and repetitive motor acts were measured on postsurgery days 5, 10, 40, and 60. The rate of increase in the body weights of rats with medial septal lesions treated with GM1 was equivalent to that of controls, while untreated rats with such lesions had reduced body weights. Rats with medial septal lesions treated with GM1 also exhibited movement times and frequency of repetitious motor acts similar to those of control rats by postsurgery day 60. No differences were found in water intake between any of the groups. Rats with medial septal lesions, whether treated with GM1 or not, had equivalent frequencies of rearings that were lower than control rats. This study emphasizes that even brief regimes of GM1 administration can exert behavioral changes in brain-damaged rats well after the treatment was administered, i.e. 40-60 days after surgery.
Gangliosides exert neuritogenic and neuronotrophic effects on the central and peripheral nervous systems 11'12'21. T h e r e is considerable evidence that gangliosides facilitate behavioral recovery following d a m a g e to several brain nuclei. For instance, the GM1 ganglioside reduced hyperemotionality after septal lesions 16, r e d u c e d locomotor hyperactivity 17, and enhanced learned alternation following lesions of the entorhinal cortex. Behavioral recovery has also been e n h a n c e d following GM1 t r e a t m e n t after bilateral caudate lesions 2°, bilateral lesions of the mediofrontal cortex 2°, and unilateral vestibular lesions 15.
damage, resulting in increased behavioral recovery 9. U n d e r some experimental conditions ganglioside treatments have not proven effective. GM1 did not protect the hippocampus from d a m a g e by colchicine 22, did not stimulate striatum reinnervation by substantia nigra grafts 5, and failed to improve brightness and pattern discrimination following bilateral ablation of the visual cortex 2. These studies emphasize that GM1 treatments are not always effective in minimizing the effects of neurological damage. In general, it would a p p e a r that the ganglioside is most effective when a tract or pathway is only partially interrupted 6"t9'23.
Many of these behavioral effects were found shortly after surgery suggesting that gangliosides may reduce the severity of brain damage close to the time of insult by decreasing e d e m a 9""~, and protecting m e m b r a n e function by maintaining ( N a + , K + ) ATPase activity 9"~3. The acute effects of GM1 t r e a t m e n t may prevent or limit early m e m b r a n e dysfunction and thus protect the brain from further
In many ways the septal area represents a nexus of neuropathways and systems related to forebrain, diencephalic, and epithalamic mechanisms. It might be anticipated that lesions of this region would interrupt some of these to a greater extent than others. In our previous study of the effects of GM1 in rats with large septal area lesions, we found that the ganglioside reduced lesion-induced h y p e r e m o -
Correspondence: R.L. Isaacson, Department of Psychology, State University of New York at Binghamton, Binghamton, NY 13901, U.S.A. 0006-8993/89/$03.50 © 1989 Elsevier Science Publishers B.V. (Biomedical Division)
397 tionality but did not alter activity behaviors. Another effect of septal area lesions is a prolonged decrease in body weight. This effect is found after nearly complete septal lesions 1"24 as well as lesions affecting more medial regions 3'4'7. This study was undertaken to investigate the effects of a short-term postoperative pharmacological treatment with GM1 on the maintenance of ingestive behaviors (body weight and water intake) following small septal lesions (medial) in rats, and to determine the effects of GM1 treatment following medial septal lesions on activity and the occurrence of repetitive motor acts on postsurgical days, 5, 10, 40, and 60. Thirty-seven adult male albino rats of the Sprague-Dawley strain (Camm Research, N J), with an average weight of 312 g, were used in this experiment. Each rat was housed individually with free access to food and water. The colony room was on a 12:12 h light-dark cycle with lights on at 07.00 h with a temperature of 22 °C. Rats were individually housed in the colony room a minimum of 20 days before surgery. Their body weights (g) and water intake (ml) were recorded daily (graduated cylinder). Rats were matched on preoperative body weights and assigned to the following groups: rats with medial septal lesions and vehicle injections (Group MS; n = 9), rats with medial septai lesions injected with ganglioside GM1 (Group MSGM; n = 11), rats with control operations and vehicle injections (Group CON; n = 8), and rats with control operations injected with GM1 (Group CONGM; n = 8). The rats were anesthetized with Chloropent (2.5 ml/kg) and placed in a Kopf stereotaxic instrument. Anodal electrolytic lesions were accomplished stereotaxically using a stainless-steel electrode insulated by epoxy except for 0.5 mm at the tip. The medial septal lesion parameters were: 9.5 mm anterior the frontal zero plane, midline, and 5.5 mm below the surface of the brain with a current of 1.5 mA passed for 13 s. The incisor bar was adjusted until lambda and bregma were on the same horizontal plane. The control operation consisted of the same procedure but current was not passed through the electrode. Rats in Groups MSGM and C O N G M received intraperitoneal injections of 30 mg/kg of ganglioside GM1 (generously supplied by Fidia Research Laboratories, Abano Terms, Italy) dissolved in 0.9% saline
solution on the day of surgery and on the next 4 successive days. Groups MS and CON received intraperitoneal injections of the saline solution on these 5 days. Body weights (g) and water intake (ml) were measured daily for 44 days after surgery. On postsurgery days 5, 10, 40, and 60, rats from Group MS (n = 9), Group CON (n = 9; including one rat whose drinking data were incomplete but for whom behavioral data were added), and Group MSGM (n = 10; data from one rat whose activity data were incomplete was eliminated) were monitored (10 min) by a computerized Digescan animal activity monitor (Omnitech, Inc., model RXYZCM). The floor of the activity chamber was 41.9 c m x 41.9 cm with 30.5 cm high plexiglass walls. The activityrelated variables measured included total distance traveled (in.), movement time (s), number of movements, time spent rearing (s), number of rearing movements, and the frequency of repeated motor actions (stereotypy). Stereotypy was defined as the breaking of the same photoelectric beam more than once in less than 1 s. Approximately 12 weeks after surgery, all rats were placed under deep chloral hydrate hypnosis (400 mg/kg, i.p.) and perfused intracardially with a physiological saline solution followed by a 10% formalin-saline solution. Brains were extracted and frozen sections made at 80 pm, saving every other section through the extent of the lesion. The sections were mounted and stained with Cresyl violet to evaluate the extent of damage. Fig. 1 shows the range of the medial septal lesions from smallest to largest at the approximate middle of the anterior-posterior extent of the lesion. The lesions extended from 9.00 mm to 7.6 mm in front of vertical plane zero TM. In most cases the posterior aspects of the medial septal area were spared. Most rats sustained some damage to the anteroventral aspects of the lateral septal area and the anterior medial parolfactory area. The size and location of the lesions were equivalent for Groups MS and MSGM. A 4 x 11 mixed design analysis of variance was used for the statistical analysis. The between factor was groups (MS, CON, MSGM, CONGM) and the within factor was 11 four-day blocks of observations. The dependent variables were body weight (in g) and water intake (ml/100 g b. wt.). Fig. 2 (top)
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Fig. 1. Reconstruction of medial septal lesions through the coronal section (A8.2) at which the greatest destruction was observed. The solid black area represents the smallest lesion found, whereas the solid and the stripped area represents the largest lesion.
presents the mean body weights across the 44 days of m e a s u r e m e n t s for all groups. As is c o m m o n after most surgical interventions, all groups shared an initial d r o p in body weight for a few days afterwards that was followed by a progressive increase thereafter. This increase was significant for all groups, Fro,320 = 654, P < 0.001. There was also a significant groups × sessions interaction, F30,32o = 3.03, P < 0.001. This indicated that although the body weights increased for all groups they did not do so at the same rate. Rats in group MS had the slowest rate of weight increase, while rats with medial septal lesions treated with GM1 had body weight increments equivalent to rats in group CON. There were no significant differences in water intake/100 g for groups, F~,32 = 1.57 or groups × test-days interaction, E30.32o = 1.42. During the postoperative observation periods rats with medial septal lesions
exhibited normal emotional reactivity to handling. Activity d a t a were analyzed by a 3 × 4 (groups × test days) mixed-design analysis of variance. Fig. 2 (middle) presents the mean amounts of time the rats were moving for each group over the 4 test days. The largest decrease in m o v e m e n t time for these groups occurred from postsurgical days 5 to 10, F3,75 = 6.43, P = 0.001. A significant interaction between groups and test days revealed that G r o u p M S G M had m o v e m e n t times equivalent to controls on postsurgical day 60, while G r o u p MS had significantly lower m o v e m e n t times on this test day, F6,75 = 3.40, P = 0.005. The total distance traveled scores reflected these trends for m o v e m e n t time across groups and test days. H o w e v e r , the groups x test days interaction was not significant for total distance traveled, F6,7_~ = 1.43. G r o u p s MS and M S G M had lower frequencies of
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Fig. 2. Mean (+ S.E.M.) body weights of all groups of rats over 44 days after surgery (top), mean time spent moving in the open field (middle), and the mean time spent making repetitive movements (bottom) on postsurgery days 5, 10, 40 and 60 for all groups except the control group given GMI. Abbreviations: ms, rats with medial septal lesions given saline after surgery; con, rats receiving control operations and given saline after surgery; ms-GM1, rats with medial septal lesions treated with GM1 after surgery; con-GMl, rats with control lesions treated with GMI after surgery.
rearing behaviors than Group CON over all test days (both of the former groups averaged 16 rearings/test day compared to 27 rearings/test day for the controls), F2.25 = 3.92, P < 0.05. Rats in Groups MS and MSGM also spent less time rearing (17 s) compared to Group CON (49 s), F2,25 = 8.18, P = 0.002. Fig. 2 (bottom) presents the frequency of stereotypy for each group during the test days. There was a significant groups x test-days interaction, F6,75 = 3.41, P = 0.005. Stereotypy was lowest for Group MSGM and highest for Group CON on postsurgery day 5, F2.6s = 6.22, P < 0.01; however, by postsurgery day 60, Groups CON and MSGM had equivalent frequencies of stereotypy that were significantly higher than Group MS, F2.65 = 3.20, P < 0.05. As expected on the basis of many previous reports (for a review, see ref. 8), medial septal lesions in rats produce few obvious effects immediately after surgery. They do, however, produce subtle but long lasting changes, many of which develop during the post-traumatic interval. Some of these changes are ones that are components of the 'septal syndrome', i.e., those changes that are commonly found after destruction of the entire septal region. Certain of these changes were alleviated by our relatively short, 5 day ganglioside treatment regime. Furthermore, the benefits of the ganglioside treatment were not apparent immediately during the treatment or shortly thereafter, but developed over the 60 days the animals were studied. This indicates that the ganglioside can initiate repair or recovery processes that can proceed after the drug treatment has been discontinued and do not depend on the continued presence of the agent. In addition the enhanced recovery produced by the ganglioside was symptom specific. Activity, weight gain, and stereotypy were all 'normalized' by the drug intervention but the lesion-induced reduction in rearing was not. This pattern of results fits with the general notion that systems involved with the first 3 types of behaviors or reactions were only partially affected by the lesions but that the mechanisms related to rearing were more severely affected. It is also likely that the effectiveness of the GM1 treatment depends on both the degree to which a behavior-related neural system is damaged and the time at which the tests are made. For instance, in a
400 previous study 16, we found that rats with complete septal lesions treated with GM1 had activity levels similar to vehicle-treated rats with the same lesions
treatments have a beneficial effect on lesion-induced behaviors when the damage to tracts or systems related to the behaviors is incomplete. Beyond
when tested on postsurgery days 12 and 30. In the present study, differences between the G M l - t r e a t e d and vehicle-treated lesioned rats only appeared after
extending previous studies to the study of a different portion of the nervous system and different behaviors, our results show even a short course of
postsurgery day 40. In overall perspective, our results are in keeping with the proposals made by others 6"923 that GM1
ganglioside treatment can be effective, provided
l Beatty, W.W. and Schwartzbaum, J.S., Consummatory behavior for sucrose following septal lesions in the rat, J. Comp. Physiol. Psychol., 78 (1972) 83-90. 2 Butler, W.M., Griesbach, E., Lable, R. and Stein, D.G., Gangliosides fail to enhance behavioral recovery after bilateral ablation of the visual cortex, J. Neurosci. Res., 17 (1987) 404-409. 3 deCastro, J.M., Stoerzinger, A., Barkmeier, D. and Ellen, P., Medial septat lesions: disruption of microregulatory patterns and circadian rhythmicity in rats, J. Comp. Physiol. Psychol.. 92 (1978) 71-84. 4 deCastro, J.M. and Taggart, D., Medial septal lesions: body weight loss and its relationship to polyuria in rats, Physiol. Behav., 22 (1979) 855-859. 5 Freed, W.J., GM1 ganglioside does not stimulate reinnervation of the striatum by substantia nigra grafts, Brain Res. Bull., 14 (1985) 91-95. 6 Gradkowska, M., Skup, M., Kiedrowski, L., Calzolari, S. and Oderfeld-Nowak, B., The effect of GMI ganglioside on cholinergic and serotonergic systems in the rat hippocampus following partial denervation is dependent on the degree of fiber degeneration, Brain Research, 375 (1986) 417-422. 7 Harrell, L.E., Parsons, D.S., Peagler, A. and Barlow, S., Alterations in regulatory behaviors induced by medial septal lesions and superior cervical ganglionectomy, Brain Research, 408 (1987) 131-140. 8 Isaacson, R.L., The Limbic System, Plenum, New York, 1982. 9 Karpiak, S.E., Li, Y.S. and Mahadik, S.P., Gangliosides (GMI and AGF2) reduce mortality due to ischemia: protection of membrane function, Stroke, 18 (1987) 184187. 10 Karpiak, S.E. and Mahadik, S.P., Reduction of cerebral edema with GM1 ganglioside, J, Neurosci. Res., 12 (1984) 485-492. 11 Ledeen, R.W., Biology of gangliosides: neurogenic and neuronotrophic properties, J. Neurosci. Res., 12 (1984) 147-159. 12 Ledeen, R.W., Gangliosides of the neuron, Trends Neurosci., 8 (1985) 169-174. 13 Li, Y.S., Mahadik, S.P., Rapport, M.M. and Karpiak, S.E., Acute effects of GM1 ganglioside: reduction in both
behavioral asymmetry and loss of Na +, K +-ATP-ase after nigrostriatal transection, Brain Research, 377 (1986) 292297. 14 Pelligrino, L.J. and Cushman, A.J., A Stereotaxic Atlas of the Rat Brain, Appleton Century Crofts, New York, 1967. 15 Petrosini, L., Behavioral recovery from unilateral vestibular lesion is facilitated by GM1 ganglioside treatment, Behav. Brain Res., 23 (1987) 117-126. 16 Poplawsky, A. and Isaacson, R.L., The GMI ganglioside hastens the reduction of hyperemotionality after septal lesions, Behav. Neural. Biol., 48 (1987) 150-158. 17 Ramirez, J.J., Fass, B., Karpiak, S.E. and Steward, O., Ganglioside treatments reduce locomotor hyperactivity after bilateral lesions of the entorhinal cortex, Neurosci. Lett., 75 (1987) 283-287. 18 Ramirez, J,J., Fass, B., Kilfoil, T., Hensehel, B., Grones, W. and Karpiak, S.E., Ganglioside-induced enhancement of behavioral recovery after bilateral lesions of the entorhinal cortex, Brain Research, 414 (1987) 85-90, 19 Sabel, B.A., Dunbar, G.L., Fass, B. and Stein, D.G., Gangliosides, neuroplasticity, and behavioral recovery after brain damage. In B.E. Will, P. Schmitt and J.C. Dalrymple-Alford (Eds.), Advances in Behavioral Biology: Volume 28. Brain Plasticity, Learning, and Memory, Plenum, New York, 1985, pp. 481-493. 20 Sabel, B.A., Dunbar, G.L. and Stein, D.G., Gangliosides minimize behavioral deficits and enhance structural repair after brain injury, J. Neurosei. Res., 12 (1984) 429-443. 21 Tettamanti, G., Ledeen, R.W., Sandhoff, K., Nagai, Y. and Toffano, G., Gangliosides and Neural Plasticity, Springer, New York, 1986. 22 Tilson, H.A., Harry, G.J., Nanry, K,, Rogers, B., Peterson, J., Jensen, K. and Dyer, R., Experiential factors in the expression of hyperemotionality produced by intradentate colchicine: lack of effect of GM1 ganglioside on colchicine-induced loss of granule cells and mossy fibers, J. Neurosci. Res., 17 (1987) 410-416. 23 Toffano, G., Agnati, L.F. and Fuxe, K.G., The effect of the ganglioside GM1 on neuronal plasticity, Int. J. Devl. Neurosci., 4 (1986) 97-100. 24 Zuromski, E.S., Donovick, P.J. and Burright, R.G., Bar •pressing for illumination change in albino rats with septal lesions, J. Comp. Physiol. Psychol., 78 (1972) 83-90.
sufficient time is allowed for the ganglioside treatment to become patent.
Brain Research. 495 (19~t/) 39{~-40(I Elsevier
BRES 23673
Brief ganglioside treatment produces delayed enhancement of functional recovery after medial septal lesions A l e x P o p l a w s k y I a n d R o b e r t L. Isaacson 2 1Department of Psychology, Bloomsburg University of Pennsylvania, Bloomsburg, PA 17815 (U. S. A. ) and 2Department of Psychology, State University of New York at Binghamton, Binghamton, NY 13901 (U.S.A.) (Accepted 2 May 1989) Key words: Medial septal lesion; Ganglioside; Recovery from brain damage; Body weight; Activity; Stereotypy
The effects of a 5-day ganglioside (GMI) treatment (30 mg/kg) on body weight and water intake subsequent to medial septal lesions were evaluated for 44 consecutive days. In addition, activity, rearing, and repetitive motor acts were measured on postsurgery days 5, 10, 40, and 60. The rate of increase in the body weights of rats with medial septal lesions treated with GM1 was equivalent to that of controls, while untreated rats with such lesions had reduced body weights. Rats with medial septal lesions treated with GM1 also exhibited movement times and frequency of repetitious motor acts similar to those of control rats by postsurgery day 60. No differences were found in water intake between any of the groups. Rats with medial septal lesions, whether treated with GM1 or not, had equivalent frequencies of rearings that were lower than control rats. This study emphasizes that even brief regimes of GM1 administration can exert behavioral changes in brain-damaged rats well after the treatment was administered, i.e. 40-60 days after surgery.
Gangliosides exert neuritogenic and neuronotrophic effects on the central and peripheral nervous systems 11'12'21. T h e r e is considerable evidence that gangliosides facilitate behavioral recovery following d a m a g e to several brain nuclei. For instance, the GM1 ganglioside reduced hyperemotionality after septal lesions 16, r e d u c e d locomotor hyperactivity 17, and enhanced learned alternation following lesions of the entorhinal cortex. Behavioral recovery has also been e n h a n c e d following GM1 t r e a t m e n t after bilateral caudate lesions 2°, bilateral lesions of the mediofrontal cortex 2°, and unilateral vestibular lesions 15.
damage, resulting in increased behavioral recovery 9. U n d e r some experimental conditions ganglioside treatments have not proven effective. GM1 did not protect the hippocampus from d a m a g e by colchicine 22, did not stimulate striatum reinnervation by substantia nigra grafts 5, and failed to improve brightness and pattern discrimination following bilateral ablation of the visual cortex 2. These studies emphasize that GM1 treatments are not always effective in minimizing the effects of neurological damage. In general, it would a p p e a r that the ganglioside is most effective when a tract or pathway is only partially interrupted 6"t9'23.
Many of these behavioral effects were found shortly after surgery suggesting that gangliosides may reduce the severity of brain damage close to the time of insult by decreasing e d e m a 9""~, and protecting m e m b r a n e function by maintaining ( N a + , K + ) ATPase activity 9"~3. The acute effects of GM1 t r e a t m e n t may prevent or limit early m e m b r a n e dysfunction and thus protect the brain from further
In many ways the septal area represents a nexus of neuropathways and systems related to forebrain, diencephalic, and epithalamic mechanisms. It might be anticipated that lesions of this region would interrupt some of these to a greater extent than others. In our previous study of the effects of GM1 in rats with large septal area lesions, we found that the ganglioside reduced lesion-induced h y p e r e m o -
Correspondence: R.L. Isaacson, Department of Psychology, State University of New York at Binghamton, Binghamton, NY 13901, U.S.A. 0006-8993/89/$03.50 © 1989 Elsevier Science Publishers B.V. (Biomedical Division)
397 tionality but did not alter activity behaviors. Another effect of septal area lesions is a prolonged decrease in body weight. This effect is found after nearly complete septal lesions 1"24 as well as lesions affecting more medial regions 3'4'7. This study was undertaken to investigate the effects of a short-term postoperative pharmacological treatment with GM1 on the maintenance of ingestive behaviors (body weight and water intake) following small septal lesions (medial) in rats, and to determine the effects of GM1 treatment following medial septal lesions on activity and the occurrence of repetitive motor acts on postsurgical days, 5, 10, 40, and 60. Thirty-seven adult male albino rats of the Sprague-Dawley strain (Camm Research, N J), with an average weight of 312 g, were used in this experiment. Each rat was housed individually with free access to food and water. The colony room was on a 12:12 h light-dark cycle with lights on at 07.00 h with a temperature of 22 °C. Rats were individually housed in the colony room a minimum of 20 days before surgery. Their body weights (g) and water intake (ml) were recorded daily (graduated cylinder). Rats were matched on preoperative body weights and assigned to the following groups: rats with medial septal lesions and vehicle injections (Group MS; n = 9), rats with medial septai lesions injected with ganglioside GM1 (Group MSGM; n = 11), rats with control operations and vehicle injections (Group CON; n = 8), and rats with control operations injected with GM1 (Group CONGM; n = 8). The rats were anesthetized with Chloropent (2.5 ml/kg) and placed in a Kopf stereotaxic instrument. Anodal electrolytic lesions were accomplished stereotaxically using a stainless-steel electrode insulated by epoxy except for 0.5 mm at the tip. The medial septal lesion parameters were: 9.5 mm anterior the frontal zero plane, midline, and 5.5 mm below the surface of the brain with a current of 1.5 mA passed for 13 s. The incisor bar was adjusted until lambda and bregma were on the same horizontal plane. The control operation consisted of the same procedure but current was not passed through the electrode. Rats in Groups MSGM and C O N G M received intraperitoneal injections of 30 mg/kg of ganglioside GM1 (generously supplied by Fidia Research Laboratories, Abano Terms, Italy) dissolved in 0.9% saline
solution on the day of surgery and on the next 4 successive days. Groups MS and CON received intraperitoneal injections of the saline solution on these 5 days. Body weights (g) and water intake (ml) were measured daily for 44 days after surgery. On postsurgery days 5, 10, 40, and 60, rats from Group MS (n = 9), Group CON (n = 9; including one rat whose drinking data were incomplete but for whom behavioral data were added), and Group MSGM (n = 10; data from one rat whose activity data were incomplete was eliminated) were monitored (10 min) by a computerized Digescan animal activity monitor (Omnitech, Inc., model RXYZCM). The floor of the activity chamber was 41.9 c m x 41.9 cm with 30.5 cm high plexiglass walls. The activityrelated variables measured included total distance traveled (in.), movement time (s), number of movements, time spent rearing (s), number of rearing movements, and the frequency of repeated motor actions (stereotypy). Stereotypy was defined as the breaking of the same photoelectric beam more than once in less than 1 s. Approximately 12 weeks after surgery, all rats were placed under deep chloral hydrate hypnosis (400 mg/kg, i.p.) and perfused intracardially with a physiological saline solution followed by a 10% formalin-saline solution. Brains were extracted and frozen sections made at 80 pm, saving every other section through the extent of the lesion. The sections were mounted and stained with Cresyl violet to evaluate the extent of damage. Fig. 1 shows the range of the medial septal lesions from smallest to largest at the approximate middle of the anterior-posterior extent of the lesion. The lesions extended from 9.00 mm to 7.6 mm in front of vertical plane zero TM. In most cases the posterior aspects of the medial septal area were spared. Most rats sustained some damage to the anteroventral aspects of the lateral septal area and the anterior medial parolfactory area. The size and location of the lesions were equivalent for Groups MS and MSGM. A 4 x 11 mixed design analysis of variance was used for the statistical analysis. The between factor was groups (MS, CON, MSGM, CONGM) and the within factor was 11 four-day blocks of observations. The dependent variables were body weight (in g) and water intake (ml/100 g b. wt.). Fig. 2 (top)
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Fig. 1. Reconstruction of medial septal lesions through the coronal section (A8.2) at which the greatest destruction was observed. The solid black area represents the smallest lesion found, whereas the solid and the stripped area represents the largest lesion.
presents the mean body weights across the 44 days of m e a s u r e m e n t s for all groups. As is c o m m o n after most surgical interventions, all groups shared an initial d r o p in body weight for a few days afterwards that was followed by a progressive increase thereafter. This increase was significant for all groups, Fro,320 = 654, P < 0.001. There was also a significant groups × sessions interaction, F30,32o = 3.03, P < 0.001. This indicated that although the body weights increased for all groups they did not do so at the same rate. Rats in group MS had the slowest rate of weight increase, while rats with medial septal lesions treated with GM1 had body weight increments equivalent to rats in group CON. There were no significant differences in water intake/100 g for groups, F~,32 = 1.57 or groups × test-days interaction, E30.32o = 1.42. During the postoperative observation periods rats with medial septal lesions
exhibited normal emotional reactivity to handling. Activity d a t a were analyzed by a 3 × 4 (groups × test days) mixed-design analysis of variance. Fig. 2 (middle) presents the mean amounts of time the rats were moving for each group over the 4 test days. The largest decrease in m o v e m e n t time for these groups occurred from postsurgical days 5 to 10, F3,75 = 6.43, P = 0.001. A significant interaction between groups and test days revealed that G r o u p M S G M had m o v e m e n t times equivalent to controls on postsurgical day 60, while G r o u p MS had significantly lower m o v e m e n t times on this test day, F6,75 = 3.40, P = 0.005. The total distance traveled scores reflected these trends for m o v e m e n t time across groups and test days. H o w e v e r , the groups x test days interaction was not significant for total distance traveled, F6,7_~ = 1.43. G r o u p s MS and M S G M had lower frequencies of
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510
610
Fig. 2. Mean (+ S.E.M.) body weights of all groups of rats over 44 days after surgery (top), mean time spent moving in the open field (middle), and the mean time spent making repetitive movements (bottom) on postsurgery days 5, 10, 40 and 60 for all groups except the control group given GMI. Abbreviations: ms, rats with medial septal lesions given saline after surgery; con, rats receiving control operations and given saline after surgery; ms-GM1, rats with medial septal lesions treated with GM1 after surgery; con-GMl, rats with control lesions treated with GMI after surgery.
rearing behaviors than Group CON over all test days (both of the former groups averaged 16 rearings/test day compared to 27 rearings/test day for the controls), F2.25 = 3.92, P < 0.05. Rats in Groups MS and MSGM also spent less time rearing (17 s) compared to Group CON (49 s), F2,25 = 8.18, P = 0.002. Fig. 2 (bottom) presents the frequency of stereotypy for each group during the test days. There was a significant groups x test-days interaction, F6,75 = 3.41, P = 0.005. Stereotypy was lowest for Group MSGM and highest for Group CON on postsurgery day 5, F2.6s = 6.22, P < 0.01; however, by postsurgery day 60, Groups CON and MSGM had equivalent frequencies of stereotypy that were significantly higher than Group MS, F2.65 = 3.20, P < 0.05. As expected on the basis of many previous reports (for a review, see ref. 8), medial septal lesions in rats produce few obvious effects immediately after surgery. They do, however, produce subtle but long lasting changes, many of which develop during the post-traumatic interval. Some of these changes are ones that are components of the 'septal syndrome', i.e., those changes that are commonly found after destruction of the entire septal region. Certain of these changes were alleviated by our relatively short, 5 day ganglioside treatment regime. Furthermore, the benefits of the ganglioside treatment were not apparent immediately during the treatment or shortly thereafter, but developed over the 60 days the animals were studied. This indicates that the ganglioside can initiate repair or recovery processes that can proceed after the drug treatment has been discontinued and do not depend on the continued presence of the agent. In addition the enhanced recovery produced by the ganglioside was symptom specific. Activity, weight gain, and stereotypy were all 'normalized' by the drug intervention but the lesion-induced reduction in rearing was not. This pattern of results fits with the general notion that systems involved with the first 3 types of behaviors or reactions were only partially affected by the lesions but that the mechanisms related to rearing were more severely affected. It is also likely that the effectiveness of the GM1 treatment depends on both the degree to which a behavior-related neural system is damaged and the time at which the tests are made. For instance, in a
400 previous study 16, we found that rats with complete septal lesions treated with GM1 had activity levels similar to vehicle-treated rats with the same lesions
treatments have a beneficial effect on lesion-induced behaviors when the damage to tracts or systems related to the behaviors is incomplete. Beyond
when tested on postsurgery days 12 and 30. In the present study, differences between the G M l - t r e a t e d and vehicle-treated lesioned rats only appeared after
extending previous studies to the study of a different portion of the nervous system and different behaviors, our results show even a short course of
postsurgery day 40. In overall perspective, our results are in keeping with the proposals made by others 6"923 that GM1
ganglioside treatment can be effective, provided
l Beatty, W.W. and Schwartzbaum, J.S., Consummatory behavior for sucrose following septal lesions in the rat, J. Comp. Physiol. Psychol., 78 (1972) 83-90. 2 Butler, W.M., Griesbach, E., Lable, R. and Stein, D.G., Gangliosides fail to enhance behavioral recovery after bilateral ablation of the visual cortex, J. Neurosci. Res., 17 (1987) 404-409. 3 deCastro, J.M., Stoerzinger, A., Barkmeier, D. and Ellen, P., Medial septat lesions: disruption of microregulatory patterns and circadian rhythmicity in rats, J. Comp. Physiol. Psychol.. 92 (1978) 71-84. 4 deCastro, J.M. and Taggart, D., Medial septal lesions: body weight loss and its relationship to polyuria in rats, Physiol. Behav., 22 (1979) 855-859. 5 Freed, W.J., GM1 ganglioside does not stimulate reinnervation of the striatum by substantia nigra grafts, Brain Res. Bull., 14 (1985) 91-95. 6 Gradkowska, M., Skup, M., Kiedrowski, L., Calzolari, S. and Oderfeld-Nowak, B., The effect of GMI ganglioside on cholinergic and serotonergic systems in the rat hippocampus following partial denervation is dependent on the degree of fiber degeneration, Brain Research, 375 (1986) 417-422. 7 Harrell, L.E., Parsons, D.S., Peagler, A. and Barlow, S., Alterations in regulatory behaviors induced by medial septal lesions and superior cervical ganglionectomy, Brain Research, 408 (1987) 131-140. 8 Isaacson, R.L., The Limbic System, Plenum, New York, 1982. 9 Karpiak, S.E., Li, Y.S. and Mahadik, S.P., Gangliosides (GMI and AGF2) reduce mortality due to ischemia: protection of membrane function, Stroke, 18 (1987) 184187. 10 Karpiak, S.E. and Mahadik, S.P., Reduction of cerebral edema with GM1 ganglioside, J, Neurosci. Res., 12 (1984) 485-492. 11 Ledeen, R.W., Biology of gangliosides: neurogenic and neuronotrophic properties, J. Neurosci. Res., 12 (1984) 147-159. 12 Ledeen, R.W., Gangliosides of the neuron, Trends Neurosci., 8 (1985) 169-174. 13 Li, Y.S., Mahadik, S.P., Rapport, M.M. and Karpiak, S.E., Acute effects of GM1 ganglioside: reduction in both
behavioral asymmetry and loss of Na +, K +-ATP-ase after nigrostriatal transection, Brain Research, 377 (1986) 292297. 14 Pelligrino, L.J. and Cushman, A.J., A Stereotaxic Atlas of the Rat Brain, Appleton Century Crofts, New York, 1967. 15 Petrosini, L., Behavioral recovery from unilateral vestibular lesion is facilitated by GM1 ganglioside treatment, Behav. Brain Res., 23 (1987) 117-126. 16 Poplawsky, A. and Isaacson, R.L., The GMI ganglioside hastens the reduction of hyperemotionality after septal lesions, Behav. Neural. Biol., 48 (1987) 150-158. 17 Ramirez, J.J., Fass, B., Karpiak, S.E. and Steward, O., Ganglioside treatments reduce locomotor hyperactivity after bilateral lesions of the entorhinal cortex, Neurosci. Lett., 75 (1987) 283-287. 18 Ramirez, J,J., Fass, B., Kilfoil, T., Hensehel, B., Grones, W. and Karpiak, S.E., Ganglioside-induced enhancement of behavioral recovery after bilateral lesions of the entorhinal cortex, Brain Research, 414 (1987) 85-90, 19 Sabel, B.A., Dunbar, G.L., Fass, B. and Stein, D.G., Gangliosides, neuroplasticity, and behavioral recovery after brain damage. In B.E. Will, P. Schmitt and J.C. Dalrymple-Alford (Eds.), Advances in Behavioral Biology: Volume 28. Brain Plasticity, Learning, and Memory, Plenum, New York, 1985, pp. 481-493. 20 Sabel, B.A., Dunbar, G.L. and Stein, D.G., Gangliosides minimize behavioral deficits and enhance structural repair after brain injury, J. Neurosei. Res., 12 (1984) 429-443. 21 Tettamanti, G., Ledeen, R.W., Sandhoff, K., Nagai, Y. and Toffano, G., Gangliosides and Neural Plasticity, Springer, New York, 1986. 22 Tilson, H.A., Harry, G.J., Nanry, K,, Rogers, B., Peterson, J., Jensen, K. and Dyer, R., Experiential factors in the expression of hyperemotionality produced by intradentate colchicine: lack of effect of GM1 ganglioside on colchicine-induced loss of granule cells and mossy fibers, J. Neurosci. Res., 17 (1987) 410-416. 23 Toffano, G., Agnati, L.F. and Fuxe, K.G., The effect of the ganglioside GM1 on neuronal plasticity, Int. J. Devl. Neurosci., 4 (1986) 97-100. 24 Zuromski, E.S., Donovick, P.J. and Burright, R.G., Bar •pressing for illumination change in albino rats with septal lesions, J. Comp. Physiol. Psychol., 78 (1972) 83-90.
sufficient time is allowed for the ganglioside treatment to become patent.