Effects of postnatal protein malnutrition on learning and memory procedures

Abstract

Protein malnutrition induces structural, neurochemical and functional changes in the central nervous system leading to alterations in cognitive and behavioral development of rats. The aim of this work was to investigate the effects of postnatal protein malnutrition on learning and memory tasks. Previously malnourished (6% protein) and well-nourished rats (16% protein) were tested in three experiments: working memory tasks in the Morris water maze (Experiment I), recognition memory of objects (Experiment II), and working memory in the water T-maze (Experiment III). The results showed higher escape latencies in malnourished animals in Experiment I, lower recognition indexes of malnourished animals in Experiment II, and no differences due to diet in Experiment III. It is suggested that protein malnutrition imposed on early life of rats can produce impairments on both working memory in the Morris maze and recognition memory in the open field tests.     

Effects of early protein malnutrition and scopolamine on learning and memory in the Morris water maze

The present study investigated the effects of early protein malnutrition on the spatial learning and memory processes. The consequences of malnutrition for the cholinergic system were evaluated by comparing the performance of malnourished and control animals in the Morris water maze after treatment with scopolamine. The learning test consisted of placing the animal in the maze to escape to a submerged platform with 12 trials per day for two consecutive days. After 24 trials, the platform was removed, the rats were placed in the maze and the time spent by them in each quadrant was recorded. After 28 days the animals were tested in a single trial to verify the retention of the spatial information. In the first Experiment, scopolamine (0.0, 0.2, 0.4 and 0.6 mg/kg per ml. i.p.) was administered 20 min before the experimental sessions. In the second experiment, a dose of 0.6 mg/kg was administered after the sessions, during the period in which learning consolidation occurs. In the first experiment, there was a significant effect of the drug, with scopolamine impairing, learning in both nutritional conditions. In the saline condition, control animals presented a better performance when compared with malnourished animals. However, 28 days later, both groups increased their latencies. With 0.2 and 0.4 mg/kg of scopolamine, the performance of both nutritional groups was similar and with 0.6 mg/kg malnourished animals performed better than controls. In the second experiment, malnourished animals were also less reactive to the effects of scopolamine, resulting in lower impairments as compared to control animals. These data suggest long-term changes in learning and memory as the result of changes produced by protein malnutrition in the cholinergic neurotransmitter system.     

Effects of early protein malnutrition and scopolamine on learning and memory in the Morris water maze

The present study investigated the effects of early protein malnutrition on the spatial learning and memory processes. The consequences of malnutrition for the cholinergic system were evaluated by comparing the performance of malnourished and control animals in the Morris water maze after treatment with scopolamine. The learning test consisted of placing the animal in the maze to escape to a submerged platform with 12 trials per day for two consecutive days. After 24 trials, the platform was removed, the rats were placed in the maze and the time spent by them in each quadrant was recorded. After 28 days the animals were tested in a single trial to verify the retention of the spatial information. In the first Experiment, scopolamine (0.0, 0.2, 0.4 and 0.6 mg/kg per ml. i.p.) was administered 20 min before the experimental sessions. In the second experiment, a dose of 0.6 mg/kg was administered after the sessions, during the period in which learning consolidation occurs. In the first experiment, there was a significant effect of the drug, with scopolamine impairing, learning in both nutritional conditions. In the saline condition, control animals presented a better performance when compared with malnourished animals. However, 28 days later, both groups increased their latencies. With 0.2 and 0.4 mg/kg of scopolamine, the performance of both nutritional groups was similar and with 0.6 mg/kg malnourished animals performed better than controls. In the second experiment, malnourished animals were also less reactive to the effects of scopolamine, resulting in lower impairments as compared to control animals. These data suggest long-term changes in learning and memory as the result of changes produced by protein malnutrition in the cholinergic neurotransmitter system.     

Brain fatty acid profiles and spatial learning in malnourished rats: effects of nutritional intervention

The aim of the present study was to determine the effects of malnutrition and nutritional rehabilitation on learning and memory performance and brain fatty acid composition. Pregnant and lactating Wistar rats were either fed ad libitum on a commercial laboratory chow or a multideficient diet from north-eastern Brazil (regional basic diet; RBD). After weaning, RBD offspring either continued on the multideficient diet (malnourished group) or switched to a control diet (rehabilitated group), until day 70. There was no difference in the passive avoidance test among the experimental groups, but malnourished rats showed important deficits in performance of the Morris water maze which were improved in the rehabilitated group. The hippocampus and cerebellum of the malnourished rats showed important changes in fatty acid profile obtained by gas-liquid chromatography, but the rehabilitated group had decreased n-3 polyunsaturated fatty acids and an increase in the proportion of arachidonic acid. The data suggest that nutritional rehabilitation results in partial restoration of fatty acid profiles and cognitive performance.     

Brain fatty acid profiles and spatial learning in malnourished rats: effects of nutritional intervention

Abstract

The aim of the present study was to determine the effects of malnutrition and nutritional rehabilitation on learning and memory performance and brain fatty acid composition. Pregnant and lactating Wistar rats were either fed ad libitum on a commercial laboratory chow or a multideficient diet from north-eastern Brazil (regional basic diet; RBD). After weaning, RBD offspring either continued on the multideficient diet (malnourished group) or switched to a control diet (rehabilitated group), until day 70. There was no difference in the passive avoidance test among the experimental groups, but malnourished rats showed important deficits in performance of the Morris water maze which were improved in the rehabilitated group. The hippocampus and cerebellum of the malnourished rats showed important changes in fatty acid profile obtained by gas-liquid chromatography, but the rehabilitated group had decreased n-3 polyunsaturated fatty acids and an increase in the proportion of arachidonic acid. The data suggest that nutritional rehabilitation results in partial restoration of fatty acid profiles and cognitive performance.     

Early protein malnutrition changes learning and memory in spaced but not in condensed trials in the Morris water-maze

Early protein malnutrition induces structural, neurochemical and functional changes in the central nervous system leading to alterations in cognitive and behavioral development of rats. The aim of the present study was to investigate the effects of protein malnutrition during lactation on acquisition and retention of spatial information using different training procedures (spaced x condensed trials). Rats treated with 16% (well-nourished) or 6% (malnourished) protein diets during the lactation phase and nutritionally recovered until 70 days of age were tested in the Morris water-maze in procedures of 1 trial/day (spaced trials), 4, 8, 12 trials/day (intermediate density) and 24 trials/day (condensed trials), completing 24 trials at the end of training. Seven and 28 days after the training the animals were tested again in just one trial to assess long-term memory. The results showed that protein malnutrition caused deficits on the spatial learning and memory in spaced but not in intermediate and condensed trials procedure. Seven and 28 days after the training there was an increase in the latency to find the platform but only malnourished animals submitted to 1 trial/day had significantly higher latency as compared with well-nourished controls. One of the possible hypotheses is that the effect protein malnutrition only in the procedure of spaced trials could be due to deficits in memory consolidation. It is suggested that these deficits can be the result of alterations produced by protein malnutrition in the hippocampal formation or in long-lasting emotional and/or motivational aspects of the rat's behavior.     

Castration Differentially Affects Spatial Working and Reference Memory in Male Rats

A male advantage for spatial learning and memory tasks is well documented among humans and rodents. A possible physiological cause for this male advantage is activational effects of androgens among males. The spatial memory of eight castrated and eight sham-castrated adult male rats was compared using a working-reference memory version of the eight-arm radial arm maze followed by a reference memory version of the Morris water maze. After maze testing, blood was collected from each rat, and testosterone levels were determined using radioimmunoassay. In the radial arm maze, castrates committed significantly more working memory errors and significantly fewer reference memory errors than did shams. In the water maze, no statistically significant differences were found for acquisition or retention. There was a trend for shams with higher testosterone levels to have better retention in the water maze, but this seemed to be due to higher levels of perseverance rather than better reference memory. Castration may have affected performance in the radial arm maze and not in the water maze because the radial arm maze was a more difficult task or because the water maze was aversively motivated while the radial arm maze was appetitively motivated. Our results indicate that androgens improve working memory and may impair reference memory, but the effects of androgens on reference memory seem to be task dependent.     

锰对大鼠空间学习记忆影响

目的探讨锰染毒大鼠空间学习记忆变化的规律。方法50只雄性大鼠适应性喂养1周后,随机分为5组,即生理盐水组(A组),Mn2+2.5,5,10,20 mg/kg(B、C、D、E组)。腹腔注射生理盐水或MnCl2溶液0.5 mL/d,共30 d。染毒结束后,采用Morris水迷宫试验测定大鼠潜伏期、总路程和穿台次数的变化。结果经多组比较的单因素方差分析,潜伏期、总路程、穿台次数各组均数间差异有统计学意义(F=7.813,P=0.000;F=8.190,P=0.000;F=3.461,P=0.000);与A组比较,B、C、D和E组潜伏期延长,差异有统计学意义(P0.05);与B组比较,D和E组潜伏期延长,差异有统计学意义(P0.05)。与A组比较,C、D和E组总路程延长,差异有统计学意义(P0.01);与B组比较,C、D和E组总路程延长,差异有统计学意义(P0.01)。与A组比较,B、C、D、E 4组穿台次数减少,差异有统计学意义(P0.05)。结论锰染毒可以引起大鼠空间学习记忆障碍。     

Spatial learning deficits induced by muscimol and CL218,872: lack of effect of prenatal malnutrition

The sensitivity of prenatal protein malnourished rats to the amnestic properties of the direct GABAA receptor agonist muscimol and the selective benzodiazepine (BZ) receptor agonist, CL218,872, was studied in the male offspring of rats provided with a protein deficient diet (6% casein) for 5 weeks prior to mating and throughout pregnancy. At postnatal day 90, rats were tested during acquisition of the submerged platform version of the Morris water maze task using four systemic doses of muscimol (0.1, 0.3, 1.0 and 1.8 mg/kg i.p.) or three systemic doses of CL218,872 (1.0, 3.2, and 5.6 mg/kg i.p.). In a dose dependent manner both drugs impaired acquisition of the task and impaired accuracy of the search pattern on the probe trial (platform removed). However, neither drug dissociated the performance of the two nutritional groups. These data are important in light of previous findings of differential behavioral effects of the non-specific BZ agonist, chlordiazepoxide (CDP), on spatial learning and on drug discrimination in prenatally malnourished rats and in the context of previous findings of reduced sensitivity to the anxiolytic effects of non-specific BZ receptor agonists across a wide variety of models of malnutrition. The present findings also support the concept that prenatal malnutrition does not affect the global functioning of the GABAA receptor, but fundamentally alters the way in which a subset of GABAA receptors (i.e. those containing the alpha2, alpha3 and/or the alpha5 but not the alpha1 subunit) is modulated by BZs.     

燃煤型氟中毒大鼠智力与脑ERK1/2表达关系

目的探讨燃煤型氟中毒大鼠智力变化及其发生机制。方法复制燃煤型氟中毒大鼠模型,用Morris水迷宫检测大鼠行为学改变;蛋白印迹及实时荧光定量PCR检测信号转导系统细胞外调节蛋白激酶1/2(ERK1/2)表达水平。结果高、低氟组大鼠逃避潜伏时间分别为(14.44±10.18)和(11.64±7.08)s,较对照组明显延长(P0.05);氟中毒大鼠第7 d穿台次数及逗留平台相限时间均较对照组减少(P0.05);染氟组大鼠脑组织中ERK1/2磷酸化蛋白较对照组升高1.5~2.3倍(P0.05),mRNA表达水平较对照组升高2.0~3.5倍(P0.05),呈剂量-效应关系。结论燃煤型氟中毒可引起大鼠学习、记忆能力降低,其机制可能与脑组织中ERK/MAPK信号转导通路的激活有关。     

The effects of perinatal protein malnutrition on spatial learning and memory behaviour and brain-derived neurotrophic factor concentration in the brain tissue in young rats

This study aimed to investigate the effects of perinatal protein malnutrition on brain derived-neurotrophic factor (BDNF) concentration in brain tissue and spatial learning and memory performance in young rats. Nine pregnant Wistar rats were assigned into three groups. Rats in one group were fed with a control diet containing 20% protein. Rats in remaining two groups were fed with a diet containing 6% protein from gestation day eight and day 15 respectively till four weeks after birth. At four weeks of age, the rat pups were evaluated for spatial learning ability using Morris Water Maze (MWM) task. At the end of the behaviour tests, rat pups were sacrificed and the brain tissue samples were collected for measurement of total protein and BDNF concentrations. It was found that rat pups fed the low protein diet had lower body weight and slightly lighter brain compared to the control pups. Total protein levels in hippocampus and cerebral cortex were significantly lower in malnourished pups than the controls. The concentration of BDNF in the hippocampus was also significantly lower in rat pups suffered protein malnutrition from early pregnancy than in the controls. MWM tests showed that perinatal protein deprivation, particularly from early pregnancy, significantly impaired learning and memory ability. The results of the present study indicate that perinatal protein malnutrition had adverse influence on spatial navigation and brain BDNF levels in rats. The decreased hippocampal BDNF concentration might partially contribute to the poor learning memory performance in the protein deprived rats.     

二硫化碳致大鼠行为改变的实验观察

本实验选用自发活动、转棒测试、步下法及Ｍｏｒｒｉｓ水迷津，观察大鼠吸入不同浓度ＣＳ２（０，３００，６００，１２００，２４００ｍｇ／ｍ＾３；４小时／日）２个月后的行为改变。结果提示，接触ＣＳ２对自发活动无明显影响，可使大鼠肌肉协调能力受损，学习记忆能力降低。     

Effect of curcumin on intracerebroventricular colchicine-induced cognitive impairment and oxidative stress in rats

This study was designed to investigate the protective effects of curcumin against colchicine-induced cognitive impairment and oxidative stress in rats. Male Wistar rats (weighing 150-200 g) received colchicine intracerebroventricularly (15 microg per rat), and cognitive dysfunctions were evaluated by the Morris water maze and the plus maze performance task and supported by biochemical tests. Central administration of colchicine caused memory deficit in both the Morris water maze and the elevated plus maze task paradigm tasks. Chronic treatment with curcumin (5-50 mg/kg, p.o.) twice daily for a period of 25 days beginning 4 days prior to colchicine injection significantly improved the colchicine-induced cognitive impairment. Biochemically, chronic administration of curcumin significantly reduced the elevated lipid peroxidation, restored the decreased reduced glutathione level and acetylcholinesterase activity, and attenuated the raised colchicine-induced elevated nitrite levels. The results of the present study indicate that curcumin has a protective role against colchicine-induced cognitive impairment and associated oxidative stress.     

母鼠妊娠前慢性不可预见性应激对子代学习记忆能力及海马突触结合蛋白I和突触结合蛋白IV表达的影响

目的 探讨母鼠妊娠前慢性不可预见性应激(chronic unpredictable stress,CUS)对1月龄子鼠学习记忆能力及大脑海马突触结合蛋白 I(synaptotagmin I,syt I)和突触结合蛋白 IV(syt IV)表达的影响。方法 SD系成年健康雌性大鼠随机分为正常对照组(8只)和CUS组(10只);采用11种刺激方法构建CUS模型;子鼠1月龄时行Morris水迷宫试验;应用Elisa法检测实验动物血清的皮质醇(cortisol,COR)和促肾上腺皮质激素释放激素(corticotropin-releasing hormone,CRH)水平;采用免疫组织化学法检测子鼠大脑海马syt I和syt IV的表达情况。 结果 1)应激结束时和分娩后,CUS组母鼠血清CRH和COR水平均高于正常对照组。2)水迷宫实验中,CUS组子鼠寻找平台潜伏期长于正常对照组子鼠,而穿越平台次数少于正常对照组子鼠,差异有统计学意义。3)1月龄CUS组子代CRH和COR水平均高于正常组子代。4)CUS组和正常对照组子鼠大脑海马syt I和syt IV蛋白表达的平均光密度(MOD)值,只在CA3层和DG层差异均有统计学意义。 结论 母鼠妊娠前CUS会使子鼠的学习记忆能力下降,并使大脑海马syt I和syt IV的表达减少。     

Malnutrition and Environmental Stimulation in Rats: Wave Latencies of the Brainstem Auditory Evoked Potentials

It has been shown that environmental stimulation may reduce the damage caused by malnutrition to morphological and behavioural parameters; however, there are no data on the effects of stimulation on the Brainstem Auditory Evoked Potentials (BAEPs). The aim of this study was to evaluate the effects of protein malnutrition, nutritional recovery and environmental stimulation on the BAEPs of the rat. On the first day of life, the animals were divided into Well-nourished (W) and Malnourished (M) groups. At weaning, half the M rats were submitted to nutritional recovery (R) until the test day. All groups were subdivided into Stimulated (S) and Non-Stimulated (N) rats. BAEPs was tested in animals exposed to clicks of 90, 80 and 70 dB of intensity. The BAEPs latencies of waves I, II, III and IV in the left ear were analysed in independent groups of rats on the 14th, 18th, 22nd, 32nd, and 42nd days of age. Statistical analysis showed diet and environmental stimulation interaction on the latencies of waves I, II, III and IV at all tested ages. WN rats showed longer latencies of waves I, II, III and IV than WS rats, and MN rats also showed longer latencies of these waves compared to WN, MS and RN at all tested ages. The results showed that malnutrition caused a delay in the latency of all BAEPs waves in rats of all ages. However, environmental stimulation reduced these latencies, reversing some damage caused by malnutrition. These data suggest that the auditory brainstem pathway is vulnerable to nutritional insults, and its structures show plasticity with environmental stimulation.     

Malnutrition and environmental stimulation in rats: wave latencies of the brainstem auditory evoked potentials

It has been shown that environmental stimulation may reduce the damage caused by malnutrition to morphological and behavioural parameters; however, there are no data on the effects of stimulation on the Brainstem Auditory Evoked Potentials (BAEPs). The aim of this study was to evaluate the effects of protein malnutrition, nutritional recovery and environmental stimulation on the BAEPs of the rat. On the first day of life, the animals were divided into Well-nourished (W) and Malnourished (M) groups. At weaning, half the M rats were submitted to nutritional recovery (R) until the test day. All groups were subdivided into Stimulated (S) and Non-Stimulated (N) rats. BAEPs was tested in animals exposed to clicks of 90, 80 and 70 dB of intensity. The BAEPs latencies of waves I, II, III and IV in the left ear were analysed in independent groups of rats on the 14th, 18th, 22nd, 32nd, and 42nd days of age. Statistical analysis showed diet and environmental stimulation interaction on the latencies of waves I, II, III and IV at all tested ages. WN rats showed longer latencies of waves I, II, III and IV than WS rats, and MN rats also showed longer latencies of these waves compared to WN, MS and RN at all tested ages. The results showed that malnutrition caused a delay in the latency of all BAEPs waves in rats of all ages. However, environmental stimulation reduced these latencies, reversing some damage caused by malnutrition. These data suggest that the auditory brainstem pathway is vulnerable to nutritional insults, and its structures show plasticity with environmental stimulation.     

Gastrodia elata Bl. Attenuated learning deficits induced by forced-swimming stress in the inhibitory avoidance task and Morris water maze

This study adopted the forced-swimming paradigm to induce depressive symptoms in rats and evaluated the effects on learning and memory processing. Furthermore, the effects of the water extract of Gastrodia elata Bl., a well-known Chinese traditional medicine, on amnesia in rats subjected to the forced-swimming procedure were studied. Rats were subjected to the forced-swimming procedure, and the inhibitory avoidance task and Morris water maze were used to assess learning and memory performance. The acquisition of the two tasks was mostly impaired after the 15-minute forced-swimming procedure. Administration of the water extract of G. elata Bl. for 21 consecutive days at a dosage of 0.5 or 1.0?g/kg of body weight significantly improved retention in the inhibitory avoidance test, and the lower dose showed a better effect than the higher one and the antidepressant fluoxetine (18?mg/kg of body weight). In the Morris water maze, the lower dose of the water extract of G. elata Bl. significantly improved retention by shortening escape latency in the first test session and increasing the time in searching the target zone during the probe test. These findings suggest that water extracts of G. elata Bl. ameliorate the learning and memory deficits induced by forced swimming.     

噪声对幼鼠学习记忆影响

目的 观察噪声对幼鼠学习记忆能力的影响.方法 将48只SD幼鼠,随机分为噪声组和对照组;噪声组在80 dB(A)噪声下持续暴露1个月,用Morris水迷宫和大鼠穿梭箱测试幼鼠学习记忆能力.结果 在4 d Morris水迷宫定航实验中,噪声组幼鼠的平均逃避潜伏期分别为(57.24±20.11),(39.28±11.23),(20.18±6.23),(9.06±2.98)s,对照组幼鼠的平均逃避潜伏期分别为(39.26±14.86),(25.61±6.77),(12.73±4.87),(4.11±1.12)s,差异均有统计学意义(P<0.01);平均有效搜索策略百分比分别为20.35%,25.32%,30.13%,38.83%,均低于对照组(P<0.05),噪声组幼鼠空间搜索实验中第1次寻找平台的潜伏期,较对照组延长(P<0.01);穿越平台的次数以及在平台象限停留的时间,均少于对照组(P<0.01);与对照组比较,噪声组幼鼠穿梭箱实验主动回避反应率降低、被动回避反应潜伏期延长(P<0.01).结论 噪声能降低幼鼠的学习记忆能力.     

Spatial Learning Deficits Induced by Muscimol and CL218,872: Lack of Effect of Prenatal Malnutrition

Abstract

The sensitivity of prenatal protein malnourished rats to the amnestic properties of the direct GABA<PRE>A</PRE> receptor agonist muscimol and the selective benzodiazepine (BZ) receptor agonist, CL218,872, was studied in the male offspring of rats provided with a protein deficient diet (6% casein) for 5 weeks prior to mating and throughout pregnancy. At postnatal day 90, rats were tested during acquisition of the submerged platform version of the Morris water maze task using four systemic doses of muscimol (0.1, 0.3, 1.0 and 1.8 mg/kg i.p.) or three systemic doses of CL218,872 (1.0, 3.2, and 5.6 mg/kg i.p.). In a dose dependent manner both drugs impaired acquisition of the task and impaired accuracy of the search pattern on the probe trial (platform removed). However, neither drug dissociated the performance of the two nutritional groups. These data are important in light of previous findings of differential behavioral effects of the non-specific BZ agonist, chlordiazepoxide (CDP), on spatial learning and on drug discrimination in prenatally malnourished rats and in the context of previous findings of reduced sensitivity to the anxiolytic effects of non-specific BZ receptor agonists across a wide variety of models of malnutrition. The present findings also support the concept that prenatal malnutrition does not affect the global functioning of the GABA<PRE>A</PRE> receptor, but fundamentally alters the way in which a subset of GABA<PRE>A</PRE> receptors (i.e. those containing the 2, 3 and/or the 5 but not the 1 subunit) is modulated by BZs.     

Short-Term Social Isolation Does Not Reduce Elevated Plus-Maze Exploration in Early Protein Malnourished Rats

An increased number of visits and time spent in the open arms of the elevated plus-maze by malnourished rats has been used as indicative of lower anxiety or higher impulsiveness. In order to study how this behavior profile responds to an anxiogenic procedure (short-term social isolation), control (16% protein) and malnourished (6% protein) rats were socially isolated prior to the test in the maze. Litters (dam plus 6 male and 2 female pups) were fed the diets from birth to 49 days of age. From 50 days on, all rats were fed a lab chow diet. Social isolation consists in removing the rats from the group and placing them in individual cages for 2 h before the test. During the test each rat was individually placed on the center of the maze and allowed to explore for 5 min. The results showed higher open arms exploration and lower attempts to enter open arms by the malnourished rats than by the controls. Social isolation decreased open arm exploration and increased time spent on the central platform in control animals, but had no effect on the malnourished rats. The results reinforce the lower anxiety or higher impulsiveness of malnourished rats, as well as the anxiogenic effect of social isolation in control rats. However, the malnourished rats were unresponsive to the anxiogenic effects of social isolation, indicating that protein deficiency early in life not only induces lower anxiety or higher impulsiveness in the maze, but also changes the behavior of these animals in response to another environmentally-induced procedure of anxiety (social isolation).