成年期双酚A暴露对小鼠行为的影响

探究成年期环境雌激素双酚A(bisphenol A, BPA)暴露对不同性别小鼠行为的影响。出生5周的小鼠灌胃染毒BPA (40、400 µg/kg/d), 同时设对照组。染毒8周后, 以开场行为检测小鼠的自发活动及探究行为, 以高架十字迷宫检测小鼠的焦虑和探究行为, 以水迷宫检测小鼠的空间学习记忆行为, 以跳台检测小鼠的被动回避记忆行为。结果显示, BPA暴露不影响同性别小鼠在开场中的各种行为, 但消除了悬空站立和扶壁站立频率的性别差异。高架十字迷宫行为测试中, BPA暴露显著减少雄鼠进入开放臂的次数和停留时间(p<0.05和p<0.01), 却显著增加雌鼠进入开放臂的次数和停留时间(p<0.05和p<0.01); 同时, BPA暴露显著减少雄鼠(p<0.05和p<0.01)却增加雌鼠在中央区的探头次数(p<0.01), 消除甚至反向诱导成年小鼠焦虑行为和探究行为的性别分化。BPA (40 µg/kg/d)暴露显著延长成年雄鼠在水迷宫中搜寻平台的平均距离(p<0.05), 但对雌鼠没有影响, BPA因此消除了正常成年小鼠空间记忆行为的性别差异。BPA(40 µg/kg/d)暴露显著缩短了雄鼠在遭电击24h后跳下平台的潜伏期(p<0.05), 但对雌鼠没有明显影响, 并因此诱导成年小鼠被动回避行为的性别差异。以上结果表明, 成年期BPA 暴露可性别特异性地影响小鼠的多种行为, 干扰成年小鼠行为的性别差异。     

环境雌激素双酚A对成年小鼠学习记忆和突触结构的影响*

双酚A (bisphenol, BPA)是一种广泛存在的环境内分泌干扰物,它可与雌激素受体结合干扰内源性雌激素对中枢神经系统的调控作用。本研究通过将10周龄小鼠灌胃染毒BPA (0.4、4、40 mg/kg/day)3个月,研究长期BPA暴露对成年小鼠记忆行为和突触可塑性的影响。开场行为测试结果表明, BPA (0.4、4、40 mg/kg/day)增加雄性的站立次数和理毛频率, BPA (4 mg/kg/day)却显著减少雌鼠的站立次数。水迷宫和被动回避行为模型检测显示, BPA主要损伤雄鼠的空间学习记忆和被动回避记忆。通过制备海马CA1区超薄切片后,电镜观测发现, BPA (0.4、40 mg/kg/day)暴露降低雄鼠海马CA1区突触数密度,缩短雄鼠突触前活性带长度,减小雄鼠突触后致密体(PSD)厚度,增加雄鼠突触间隙宽度。进一步用Western blot方法检测突触前、后的标志性蛋白Synapsin I和PSD95以及兴奋性氨基酸NMDA受体NR1亚基和AMPA受体GluR1亚基蛋白的表达,发现BPA暴露致雄鼠Synapsin I、PSD95、NR1蛋白表达水平下调。而BPA对雌鼠的记忆行为、突触形态、突触蛋白和受体蛋白均没有明显作用。以上结果提示长期B PA暴露性别特异性地影响成年小鼠的活动性和探究行为,损伤雄鼠的学习记忆,这些作用可能通过下调突触蛋白和NMDA受体的表达而负性影响突触结构可塑性,最终影响雄鼠的学习记忆功能。     

生命早期双酚A暴露对子代成年小鼠焦虑和抑郁行为的影响及其神经机制

双酚A (bisphenol A, BPA)对脑和行为发育的影响已引起关注, 本研究探讨围生期不同发育阶段母体BPA暴露对仔鼠成年后焦虑和抑郁行为的影响。分别在妊娠期(妊娠第7天~出生)和哺乳期(出生第1~14天) 将母鼠暴露于BPA (0.4、4 mg/kg/day), 以旷场、明暗箱、镜子迷宫、高架十字迷宫等多种模型检测生后56天(postnatal day 56)仔鼠的焦虑行为, 以强迫游泳模型检测其抑郁行为。结果显示, 妊娠期BPA暴露的成年雌性仔鼠在所有4种模型中均检测到促焦虑作用, 而哺乳期BPA暴露的雌鼠、妊娠期或哺乳期BPA暴露的雄鼠仅在2种模型中检测到促焦虑作用。妊娠期BPA暴露显著加重雌雄仔鼠的抑郁行为, 而哺乳期仅高剂量BPA加重仔鼠的抑郁行为。进一步的Western blot分析显示, 妊娠期或哺乳期BPA暴露显著下调成年后雌雄仔鼠海马和杏仁核AMPA受体GluR1亚基的表达, 但对NMDA受体NR1亚基的影响不一致。以上结果提示, 妊娠期或哺乳期BPA暴露对成年雌雄仔鼠均有不同程度的促焦虑和抑郁作用, 其中妊娠期暴露对雌鼠的作用最显著, 海马和杏仁核AMPA受体活动的减弱可能与围生期BPA暴露加重仔鼠成年后的焦虑和抑郁行为有关。     

长期双酚A暴露对成年小鼠恐惧记忆的影响

双酚A (bisphenol, BPA)是一种广泛应用于塑料制品的环境内分泌干扰物, 具有弱雌激素和抗雄激素活性, 与雌激素受体有一定的亲和力。本实验探讨长期BPA暴露对成年小鼠恐惧记忆的影响及其神经机制。将9周龄雄性小鼠暴露于BPA (0.4、4、40 mg/kg/d) 90 d, 建立小鼠亚慢性BPA暴露模型后, 进行场景性条件恐惧训练, 然后分别在电击后1 hr及24 hr检测小鼠的僵立时间, 同时在电击前、电击后1 hr及24 hr检测海马相关蛋白表达变化。结果表明, BPA (4、40 mg/kg/d)暴露延长小鼠场景性条件恐惧训练后1 hr及24 hr的僵立时间。Western blot蛋白检测结果显示, 行为训练前, BPA降低了小鼠NMDA受体NR1亚基表达水平, 上调组蛋白去乙酰化酶2表达。行为训练后1 hr及24 hr, BPA促进海马NMDA受体亚基NR1和组蛋白H3乙酰化表达, 抑制组蛋白去乙酰化酶2表达, 同时促进ERK1/2磷酸化水平。以上结果表明, 长期BPA暴露提高成年小鼠恐惧记忆获得的同时延长恐惧记忆的保持; 该作用可能通过激活海马的ERK1/2通路而改变核内组蛋白乙酰化和NMDA受体水平进行。     

脂多糖免疫激活导致抑郁性行为的动物研究：剂量和时程效应

“抑郁症细胞因子假说”的提出为抑郁障碍的病因学研究提供了一个新的方向,为了探讨脂多糖（lipopolyaccharide,LPS）诱导的免疫激活与抑郁性行为产生之间的关系,本研究采用50只SD大鼠随机分为五组LPS400,LPS200,LPS50,LPS10,LPS0,分别于实验期第0天和第3天注入LPS400ug/kg,,200ug/kg,50ug/kg,10ug/kg和生理盐水。以糖精水偏爱,旷场行为和高架十字迷宫评定大鼠LPS注射后2小时,24小时,48小时的行为变化。结果显示一次LPS注射后2小时,LPS50,LPS200,LPS400组动物与生理盐水组动物相比较,其糖精水偏爱分数（p<0.01）,旷场中的水平活动距离(p<0.01)和直立行为(p<0.01)以及高架十字迷宫中的闭合臂进入次数(p<0.01)和开放臂进入次数显著下降(p<0.01);重复注射后2小时LPS注射组动物的闭合臂进入次数显著降低（p<0.01）;但LPS10组与生理盐水组动物在行为上没有差异,50ug/kg,200ug/kg和400ug/kg剂量的各组之间没有差异。LPS注射后24小时和48小时以及重复注射后大鼠的行为没有发现显著变化。提示LPS诱导的免疫激活对抑郁行为产生有一定的作用。免疫激活的细胞因子能够导致动物出现明显的抑郁性行为,但是这种行为缺乏长时程效应,因此LPS诱导的抑郁障碍的动物模型应用是非常有限的。免疫激活的前炎性细胞因子可能是导致抑郁障碍产生的其中一个原因而不是唯一原因。     

慢性应激对大鼠空间学习记忆及海马和前脑皮层突触体膜流动性的影响

慢性应激对学习记忆功能的影响是神经科学的热点问题, 在脑内, 海马和前额叶是与学习记忆功能密切相关的重要脑区, 也是应激易累及损伤的主要靶区。膜流动性的改变在神经细胞功能活动中起重要作用。为探讨慢性应激对大鼠空间学习记忆功能的影响及前脑皮层和海马突触体膜流动性的作用。采用多因素慢性应激动物模型, 通过开场试验和Morris水迷宫测试大鼠行为及空间学习记忆能力; 并且测定大鼠前脑皮层和海马突触体膜流动性和突触体内游离Ca2+浓度的变化。研究结果显示, 与对照组相比, 应激组大鼠在应激后即刻, 在新异环境中的自发活动和探究行为显著降低(p<0.05, p<0.01), 空间学习记忆能力明显下降(p<0.05, p <0.01); 并且应激组大鼠前脑皮层和海马突触体膜流动性显著降低(p <0.05, p <0.01); 而突触体内游离Ca2+浓度的显著增加 (p <0.05, p <0.01)。停止应激后一周, 应激大鼠的各项指标有所恢复, 但仍未达到正常水平。研究结果提示, 慢性应激引起大鼠明显的开场行为改变和空间学习记忆功能障碍, 这些变化可能与突触体膜流动性和突触体内游离Ca2+浓度的变化密切相关。     

高压氧对小鼠学习记忆及脑细胞形态结构的影响

实验用两种行为模型（旷场行为模型和Ｙ－迷宫分辨学习模型）观察了幼龄小鼠在不同压力的高压氧处理后,对新异环境的探究行为和自发活动情况,以及学习记忆能力的变化;并用ＸＹ－生物医学电脑图像分析仪分析了与学习记忆相关的脑区（大脑皮层、海马）神经元密度,细胞核面积,胞核／胞浆比值的变化。结果表明：（１）与对照组相比,吸０.１ＭＰａ高压氧的幼鼠学习记忆能力明显提高,相关脑区的神经元密度、细胞核面积、胞核／浆比均显著增加。（２）吸０.２５ＭＰａ高压氧的幼鼠学习记忆能力与对照组相比无明显变化,但其在新异环境中的自发行为明显减少。提示：慢性吸入０.１ＭＰａ高压氧有利于促进幼鼠脑的生长发育,增强脑功能活动。     

雄性HSF1基因缺陷小鼠的行为改变

为研究HSF1基因缺陷小鼠的行为特征,探索HSF1基因在小鼠行为表现中的作用。选取6~7个月大雄性HSF1基因缺陷小鼠39只及野生型小鼠36只进行情绪性评分、旷场实验、高架十字迷宫实验、简易迷津实验、T-CAT实验、独木桥实验和悬挂实验以观察其情绪性唤醒水平、焦虑水平、探索行为、工作记忆能力和运动能力。结果表明HSF1基因缺陷小鼠的情绪唤醒水平和焦虑水平较低、探索行为减少、T-CAT中转换率较低,提示小鼠的情绪、探索动机和工作记忆受HSF1基因的调控     

东莨菪碱对吗啡诱导的大鼠行为敏感化的影响

药物重复处理导致的行为敏感化与成瘾过程密切相关。本实验检验东莨菪碱对吗啡诱导的大鼠行为敏感化发展和转化的影响。实验一动物分为3组,分别进行生理盐水（对照组）、吗啡（10mg/kg,吗啡组）、吗啡（10mg/kg）＋东莨菪碱（3mg/kg,吗啡-东莨菪碱组）前处理,36小时腹腔注射4次（第1~2天）。自然戒断7天（第3~9天）。第10天,所有动物均使用吗啡（4mg/kg）激发,记录动物的自发活动量;第24天,吗啡组和吗啡-东莨菪碱组动物重复第10天的操作。实验二动物分为3组,分别接受生理盐水（对照组）、吗啡（10mg/kg,吗啡组）、吗啡（10mg/kg,吗啡-东莨菪碱组）处理,36小时腹腔注射4次（第1~3天）;间隔12小时后,3组动物分别接受生理盐水、生理盐水和东莨菪碱（3mg/kg）处理,仍为36小时腹腔注射4次（第3 ~5天）。第6~9天不进行药物处理。第10天和第17天,分别使用吗啡（4mg/kg）激发,记录动物的活动量。记录时间均为两小时(10分钟为一个记录单元)。结果表明,东莨菪碱能够抑制吗啡诱导的行为敏感化的发展,一定程度上也能够延缓行为敏感化的转化但没有阻断这种转化     

毒扁豆碱对吗啡导致的大鼠行为敏感化的抑制作用

药物滥用导致的行为敏感化被认为与成瘾过程密切相关。本实验探讨吗啡导致的大鼠行为敏感化与神经递质乙酰胆碱的关系。实验动物分为3组,分别进行生理盐水、吗啡（10.0mg/kg）、吗啡（10.0mg/kg）＋胆碱酯酶抑制剂毒扁豆碱（0.2mg/kg）前处理,36小时腹腔注射4次。前处理结束1周所有动物注射小剂量吗啡（4.0mg/kg）;使用生理盐水前处理的动物,第2周注射毒扁豆碱（0.2mg/kg）;使用吗啡前处理的动物,第2周注射小剂量吗啡＋毒扁豆碱（0.2mg/kg）,第3周再次注射小剂量吗啡。动物每次接受注射后立即记录其在两小时内的活动量(10分钟为一个记录单元)。结果表明,毒扁豆碱既能够抑制吗啡诱导的行为敏感化,也能够阻断小剂量吗啡对行为敏感化的“点燃”作用。由此推论,吗啡导致的行为敏感化与其抑制乙酰胆碱分泌有关。     

Immune stress in late pregnant rats decreases length of gestation and fecundity, and alters later cognitive and affective behaviour of surviving pre-adolescent offspring

Immune challenge during pregnancy is associated with preterm birth and poor perinatal development. The mechanisms of these effects are not known. 5α-Pregnan-3α-ol-20-one (3α,5α-THP), the neuroactive metabolite of progesterone, is critical for neurodevelopment and stress responses, and can influence cognition and affective behaviours. To develop an immune challenge model of preterm birth, pregnant Long-Evans rat dams were administered lipopolysaccharide [LPS; 30?μg/kg/ml, intraperitoneal (IP)], interleukin-1β (IL-1β; 1?μg/rat, IP) or vehicle (0.9% saline, IP) daily on gestational days 17-21. Compared to control treatment, prenatal LPS or IL-1β reduced gestational length and the number of viable pups born. At 28-30 days of age, male and female offspring of mothers exposed to prenatal IL-1β had reduced cognitive performance in the object recognition task compared to controls. In females, but not males, prenatal IL-1β reduced anxiety-like behaviour, indicated by entries to the centre of an open field. In the hippocampus, progesterone turnover to its 5α-reduced metabolites was lower in prenatally exposed IL-1β female, but not in male offspring. IL-1β-exposed males and females had reduced oestradiol content in hippocampus, medial prefrontal cortex and diencephalon compared to controls. Thus, immune stress during late pregnancy reduced gestational length and negatively impacted birth outcomes, hippocampal function and central neurosteroid formation in the offspring.     

Effects of Early Maternal Separation on Subsequent Reproductive and Behavioral Outcomes in Male Rats

To investigate effects of maternal separation on reproductive and behavioral outcomes, male Wistar rats were separated from their mothers daily for 3 hr (maternal separation; MS) or 0 hr (control) from postnatal day (PND) 1 to 14. Timing of puberty, reproductive parameters, hormone levels, and aggressive behaviors in juvenile and adult rats were examined. Contrary to expectations, there was no effect of maternal separation on any measure of aggression. However, maternal separation altered peripubertal testosterone secretion and increased mean day of preputial separation. In addition, adult MS males demonstrated less total sexual behavior. There was no difference in sperm counts or testosterone levels at necropsy on PND 56 or in adulthood, but seminal vesicle weights were increased in adult MS rats. These results suggest that early life stress may influence hypothalamic-pituitary-gonadal axis development in males, at least during peripuberty.     

Male versus female Siamese fighting fish as reinforcing stimuli for conspecific males in single- and two-choice operant situations

Two experiments attempted to determine the reinforcing effectiveness of visual exposure to a female versus a male stimulus in male Siamese fighting fish (Betta splendens). This was accomplished by making male or female presentation contingent upon an operant ring swimming response in single- (Experiment 1) and two-choice (Experiment 2) continuous reinforcement operant situations for 1 hr per day. It was found that visual exposure to a female stimulus was behaviorally reinforcing in a way comparable to that typically found in intermale studies. If given a choice, male test subjects were observed to demonstrate a consistent preference for the female as opposed to the male stimulus. The results are discussed in terms of stimulus discrimination and approach-avoidance conflict as differentially elicited by male and female target stimuli.     

Effects of prenatal ethanol exposure on juvenile play-fighting and postpubertal aggression in rats

The effects of prenatal ethanol exposure on juvenile play-fighting and postpubertal aggressive behavior in rats were longitudinally assessed in the context of more conventionally applied physical and behavioral measures. Pregnant animals were treated with either 2 gm/kg/day ethanol or isocaloric sucrose over gestation Days 6-19. Reproduction and somatic variables included maternal weight over gestation, offspring weight over Days 1-90, and age at eye opening and incisor eruption. Behavioral variables consisted of negative geotaxis, olfactory discrimination, activity, juvenile play-fighting, and postpubertal aggression. Ethanol offspring had lower birth weights, but there was no significant prenatal treatment effect on subsequent offspring weights or on any other reproductive or somatic variable. Both male and female ethanol-exposed offspring exhibited more play-fighting responses when paired with same-sex controls. Postpubertal aggression levels were assessed in males only. Ethanol-exposed offspring were more aggressive than controls and there was a significant positive correlation between play-fighting and postpubertal aggression ranks. No other behavioral measures discriminated between prenatal treatment groups and none were significantly correlated with either play-fighting or postpubertal aggression rank. The results are consistent with the position that juvenile play-fighting and postpubertal aggression are subserved by common substrates. They also are consistent with predictions derived from the hypothesis concerning a response-inhibition deficit as an effect of prenatal ethanol exposure on behavior.     

Effect of exposure to lithium-paired or amphetamine-paired saccharin solution on open arm avoidance in an elevated plus maze

Recent evidence suggests that drug-induced conditioned taste avoidance may be mediated by conditioned fear (e.g., Parker, 2003). The experiments reported here evaluated the effect of exposure to a drug-paired flavor on open arm exploration in an elevated plus maze (EPM), a measure of fear. When rats were tested on a familiar (trial 2) EPM, but not on a novel (trial 1) EPM, prior exposure to a lithium-paired saccharin solution enhanced open arm activity relative to saline-paired saccharin. On the other hand, when rats were exposed to lithium-paired saccharin during plus maze exposure, they displayed suppressed open arm activity relative to unpaired controls when tested in a familiar maze. The pattern of results was specific to the conditional affective properties of the taste, because exposure to unconditional sickness produced by administration of lithium, and unconditionally unpalatable quinine solution did not produce this pattern. These results were interpreted in terms of the opponent process model of motivation; that is, exposure to a lithium-paired flavor elicits conditioned fear which is immediately followed by conditioned relief when the exposure is terminated. On the other hand, exposure to an amphetamine-paired flavor either before or during EPM testing enhanced open arm exploration. Since the strength of taste avoidance did not differ among amphetamine and lithium conditioned rats, these results provide further evidence that the nature of a saccharin-lithium association differs from that of a saccharin-amphetamine association.     

Sex differences in spatial and non-spatial Y-maze performance after chronic stress

Chronic restraint is known to alter hippocampal CA3 dendritic morphology and spatial memory in male rats. The present study examined whether female rats, which exhibit different anatomical adaptations to chronic stress than those of males, would also show spatial memory impairments. Male and female Sprague-Dawley rats were restrained for 6 h/day for 21 days, a time frame previously demonstrated to cause hippocampal CA3 dendritic atrophy. The day after the last restraint session, rats were tested on a Y-maze, a habituation task that can be used to assess spatial memory. Chronic stress impaired Y-maze performance in both sexes without affecting levels of locomotion as measured by total arm entries in the first minute. However, Y-maze performance of stressed females improved in 2-5 min when chronically stressed males continued to show poor Y-maze performance. The enhanced Y-maze performance of chronically stressed females occurred when total arm entries were higher compared to the entries made by males. Therefore, correlations were performed between total arm entries and spatial memory in 1 and 2-5 min. In the first minute when control females demonstrated functional spatial memory, female controls with the lowest locomotor levels exhibited the best performance. The correlations for stressed females were not significant, and neither were the correlations for any group in 2-5 min. Overall, these results show important sex differences in response to chronic stress with females exhibiting an ability to recover quickly from deficits in Y-maze performance.     

Rat gestation during space flight: Outcomes for dams and their offspring born after return to earth

Sprague-Dawley rats were studied to learn whether gestation in the near-zero gravity, high radiation environment of space impacts selected mammalian postnatal events. Ten rats spent days nine to twenty of pregnancy aboard the space shuttle orbiterAtlantis (STS-66). Their movement, was studied shortly after return to Earth; subsequently, several of their offspring were cross-fostered and examined through postnatal day 81 (P81) for whole body growth and somatic motor development. Values for the flight animals were compared to ground-based control groups. Relative to controls, the pregnant flight rats showed a marked paucity of locomotion during the first few hours after returning to Earth. There was greater likelihood of perinatal morbidity for the offspring of flight dams when compared to the control groups. Whole body weight of surviving offspring, averaged for each group separately, showed typical sigmoidal growth curves when plotted against postnatal age. The flight group for our study had a larger ratio of female to male pups, and that was sufficient to account for the lower average daily weight gained by the flight animals when compared to the control groups. Walking was universally achieved by P13 and preceded eye opening which was complete in all pups by P17. Thus, both of these developmental horizons were attained on schedule in the flight as well as the control rats. Characteristic changes were observed in hind limb step length and gait width as the pups grew. These patterns occurred at the same time in each group of rats. Therefore, prenatal space flight from days nine to twenty of gestation did not interfere with the establishment of normal patterns for hind paw placement during walking.     

Predictor variables for a 100-km race time in male ultra-marathoners

In 169 male 100-km ultra-marathoners, the variables of anthropometry, training, and prerace experience, in order to predict race time, were investigated. In the bivariate analysis, age (r = .24), body mass (r = .20), Body Mass Index (r = .29), circumference of upper arm (r = .26), percent body fat (r = .45), mean weekly running hours (r = -.21), mean weekly running kilometers (r = -.43), mean speed in training (r=-.56), personal best time in a marathon (r = .65), the number of finished 100-km ultra-runs (r = .24), and the personal best time in a 100-km ultra-run (r = .72) were associated with race time. Stepwise multiple regression showed that training speed (p < .0001), mean weekly running kilometers (p < .0001), and age (p < .0001) were the best correlations for a 100-km race time. Performance may be predicted (n=169, r2 = .43) by the following equation: 100-km race time (min) = 1085.60 - 36.26 x (training speed, km/hr.) - 1.43 x (training volume, km/wk.) + 2.50 x (age, yr.). Overall, intensity of training might be more important for a successful outcome in a 100-km race than anthropometric attributes. Motivation to train intensely for such an ultra-endurance run should be explored as this might be the key for a successful finish.