神经元营养因子对大鼠隔—海马通路损伤的影响

为评价神经生长因子(NGF)、混合型神经节苷脂(GM)和单唾液酸神经节苷脂(GM1)对中枢胆碱能神经损伤早期的影响,在大鼠单侧隔-海马通路部分损伤后即时经脑室分别注入上述三种神经元营养因子,7d后取两侧海马分别测定乙酰胆碱(ACh)、胆碱乙酰基转移酶(ChAT)和胆碱酯酶(ChE)。损伤对照组(脑室注入盐水)术侧海马ACh含量保留率为对侧的20.3％,ChAT活力为50％,ChE活力为48.3％。给予NGF、GM或GM1的实验组,ACh含量保留率分别为34.9％,35.3％和47.7％;ChAT活力为77.4％,78.4％和69.2％;而ChE活力的保留率未见明显改变。这些神经元营养因子显著增加了大鼠隔-海马通路损伤后海马内ACh含量和ChAT活力,说明它们减轻了损伤侧海马胆碱能神经纤维的破坏,具有明显的损伤早期保护作用。     

胚胎海马伞移植物对成年大鼠海马胆碱能纤维生长的引导效应

在胚胎和新生的中枢神经系统(CNS)内,发育中的纤维束通道能引导轴突的生长。为了了解发育中的纤维束通道能否引导成年CNS轴突的生长,将胚胎海马伞移植到成年大鼠的海马,两周后,用AChE组织化学方法检查移植物内的胆碱能纤维。结果如下:在胚胎的海马伞移植物内出现大量的胆碱能纤维,但在成年的海马伞移植物内没有宿主的胆碱能纤维长入;如果在移植胚胎海马伞的同时,切断宿主的海马伞-穹窿通路,则在胚胎移植物和宿主海马内均无胆碱能纤维;将胚胎海马伞作成悬浮液进行移植,在移植部位,仅能看到少数长的胆碱能纤维;但是若把胚胎海马伞的组织碎片粘附在硝化纤维素滤纸条周围,再移植到成年大鼠海马内,来自宿主海马的大量胆碱能纤维被吸引围绕着滤纸条并沿其表面生长。结果似乎表明:胚胎海马伞或胚胎海马伞碎片都能有效引导宿主海马胆碱能纤维的生长。因此,胚胎海马伞和其它发育中的CNS纤维束通道可能是引导成年CNS轴突生长的良好天然基质。     

海马内移植颈上神经节组织改善穹窿－海马伞横切大鼠的记忆功能

３８只Ｗｉｓｔａｒ大鼠双侧穹窿－海马伞损伤后．以被动回避反应为指标观察到记忆明显受损,与损伤前相比,有记忆动物自６５．３％降至１３．６％,两者差别非常显著。于穹窿－海马伞损伤后记忆丧失动物（ｎ＝１５）自体一侧分离出颈上神经节（ＳＣＧ）,切为２－３块并在室温下孵育于２０—５０μｇ／ｍｌ２．５ｓＮＧＦ中１—２ｈ,而后移植于自体双侧海马背侧,移植４周后观察到动物记忆明显恢复,恢复记忆的动物数占７３．３％。在行为实验基础上应用荧光组化方法检查了移植细胞成活情况并测量了海马内去甲肾上腺素（ＮＡ）的含量。移植后２周海马内ＮＡ含量比损伤组有明显上升。移植后一个月,可见部分移植细胞成活并有神经纤维生长。实验表明．穹窿－海马伞损伤大鼠海马内自体移植ＳＣＧ,通过神经递质的局部补充对动物丧失的记忆能力具有一定改善作用。     

鼠脑移植恢复受损脑神经元联系的尝试成功

最近,N.Sunde等用X射线损伤新生鼠双侧海马区域,随后将另一新生鼠的正常齿状回组织移植入受损伤鼠单侧海马区域(另一侧作为对照)。发现:在非胆碱能、非单胺能神经系统,与胆碱能、单胺能系统一样,移植物和宿主神经元能够建立相互联系,并且移植物可恢复宿主已被破坏的神经联系。80%以上的齿状回颗粒细胞在出生以后形成。出生后,X射线照射几乎能选择性地使颗粒细胞数减少到正常的15%,残存的颗粒细胞传入纤维形成散乱的扁平状突起伸向CA3区维体细胞。同时苔藓纤维突起明显减少。这种X射线照射模型在许多方面与自然发生的神经细胞恶性改变及早期退行性疾病相同。将     

穹窿切断后海马神经元GR表达变化的研究

目的实施大鼠穹窿切断术研究海马神经元核受体GR(glucocorticoidreceptor糖皮质激素受体)表达的变化。方法建立大鼠穹窿切断模型,于穹窿切断术后0、4、7、10d取材;同时取材假手术组(非穹窿切断术)作为对照,应用免疫组化、WesternBlotting方法分别进行各组海马神经元GR表达变化的观察及定量检测。结果穹窿切断7d后,免疫组化和WesternBlotting结果显示海马神经元GR表达下调,10d下调更为显著。结论穹窿切断后,海马GR表达下调,提示可能减弱海马对HPA轴的抑制。     

横断大鼠穹窿-海马伞对其海马CA3区突触形态的影响

目的：观察横断大鼠穹窿-海马伞对其海马突触形态的影响。方法：横断大鼠双侧穹窿-海马伞（FF）建立动物模型,于手术前、后对大鼠进行迷宫检查,重点对海马CA3区多形层突触界面的结构参数进行定量分析。结果：突触界面曲率减小,突触间隙宽度加大,突触后膜致密物质厚度明显变薄,穿孔性突触的比例也有不同程度降低。结论：横断穹窿-海马伞引起海马CA3区突触形态明显改变,推测海马内Ach的正常水平对维持海马CA3区突触界面超微结构有重要作用。     

海马内注射6—OHDA对大鼠条件回避行为习得和保持的影响

本文研究了在大鼠两侧背海马内注射6-OHDA后,儿茶酚胺能纤维和多巴胺能纤维末梢在穿梭箱条件性回避行为习得和保持中的作用。结果表明:(1)6-OHDA使海马内多巴胺含量明显减少,特别是去甲肾上腺素含量的减少更为显著。(2)条件性回避行为的习得受到损害。(3)海马内注射加压素易化条件性回避行为的效应被阻断。     

甘麦大枣汤对抑郁模型大鼠海马神经元突触结构及结构蛋白表达的影响*

目的: 观察甘麦大枣汤对抑郁模型大鼠行为学及单胺递质的影响,并从突触结构及结构蛋白MAP-2与GAP-43表达改变探讨其潜在作用机制。方法: 60只SD大鼠随机分为5组:正常对照组、模型组、氟西汀组(10.8 mg/kg)、甘麦大枣汤高、低剂量组(9.72、4.86 g/kg),每组12只。除对照组外,其余各组大鼠均采用慢性不可预见性温和应激(CUMS)建立抑郁模型,并于造模同时给药组灌胃给药,连续21 d。采用糖水消耗实验和旷场测试评价大鼠抑郁样行为,ELISA法检测海马单胺递质5-HT、NE含量,Golgi染色观察神经元突触损伤情况,免疫组化法和Western blot法检测海马突触结构蛋白MAP-2和GAP-43的表达。结果: 与对照组比较,抑郁模型大鼠糖水偏好度及自主活动评分均显著下降(P＜0.01),海马5-HT、NE含量显著下降(P＜0.01),树突棘缺失明显,同时MAP-2和GAP-43表达均显著下调(P＜0.01);甘麦大枣汤干预后,模型大鼠抑郁样行为明显缓解(P＜0.01),5-HT、NE含量显著升高(P＜0.05),树突棘密度、长度及分枝增加,MAP-2和GAP-43表达显著回升(P＜0.01)。结论: 甘麦大枣汤能改善抑郁模型大鼠的抑郁样行为,升高海马单胺递质含量,其作用可能与上调突触结构蛋白、缓解神经元突触损伤有关。     

CNTF对烧伤大鼠血清引起大鼠海马神经元细胞毒性的影响

应用整体和离体神经元培养,观察CNTF对烧伤大鼠海马神经元及烧伤血清引起海马神经元损伤的影响,结果表明,大鼠烧伤后海马组织神经元数目减少,NO含量升高;烧伤大鼠血清可引起培养的海马神经元细胞存活率下降,培养液中NO含量升高;CNTF能降低烧伤大鼠海马组织中NO的含量,保护海马神经元,并能提高培养的海马神经元的存活率,减少培养液中NO含量,其作用呈剂量依赖性;CNTF对神经元存活率的影响与NO含量呈显著负相关,提示CNTF对烧伤大鼠血清引起的海马神经元损伤有保护作用,其作用机制可能是通过抑制NO的神经毒性。     

海马内移植颈上神经节组织改善穹窿—海马伞横切大鼠的记忆功能

３８只Ｗｉｓｔａｒ大鼠双穹窿－海马伞损伤后,以被动回避反应为指标观察到记忆明显受损,与损伤前相比,有记忆动物自６５．３％降至１３．６％,两者差别非常显著。于穹窿－海马伞损伤后记忆丧失动物（ｎ＝１５）自体一侧分离出颈上神经节（ＳＣＧ）,切为２－３块并在室温下孵育于２０－５０μｇ／ｍｌ２．５ｓＮＧＦ中１－２ｈ,而后移植于自体双侧海马背侧,移植４周后观察至动物记忆明显恢复,恢复记忆的动物数占７３．３％。     

Restoration of High Affinity Choline Uptake in the Hippocampal Formation Following Septal Cell Suspension Transplants in Rats with Fimbria-Fornix Lesions

High affinity choline uptake (HACU) was investigated in the hippocampal formation following fetal septal cell suspension transplants into rats with fimbria-fornix lesions. Nine-14 weeks after transplantation, HACU was markedly decreased in hippocampi from animals with fimbria-fornix lesions; this decrease was ameliorated by fetal septal cells transplanted into the host hippocampus. HACU related to septal transplantation was activated in vitro by K+, and in vivo by the administration of scopolamine and picrotoxin. These findings suggest that fetal septal cell transplantation can restore HACU in the host hippocampus following fimbria-fornix lesions, and that HACU related to the graft has pharmacological properties similar to those of the normal adult HACU system. The activation of HACU by picrotoxin, a gamma-aminobutyric acid (GABA) antagonist, suggests that transplanted cholinergic neurons receive either direct or indirect functional input from GABAergic afferents from the transplant and/or host hippocampus. Lesions of the fimbria-fornix also resulted in an increased binding to muscarinic receptors in the dorsal hippocampus. This increase in binding was not significantly ameliorated by intrahippocampal grafts of cholinergic neurons.     

Artificial Theta Stimulation Impairs Encoding of Contextual Fear Memory

Several experiments have demonstrated an intimate relationship between hippocampal theta rhythm (4–12 Hz) and memory. Lesioning the medial septum or fimbria-fornix, a fiber track connecting the hippocampus and the medial septum, abolishes the theta rhythm and results in a severe impairment in declarative memory. To assess whether there is a causal relationship between hippocampal theta and memory formation we investigated whether restoration of hippocampal theta by electrical stimulation during the encoding phase also restores fimbria-fornix lesion induced memory deficit in rats in the fear conditioning paradigm. Male Wistar rats underwent sham or fimbria-fornix lesion operation. Stimulation electrodes were implanted in the ventral hippocampal commissure and recording electrodes in the septal hippocampus. Artificial theta stimulation of 8 Hz was delivered during 3-min free exploration of the test cage in half of the rats before aversive conditioning with three foot shocks during 2 min. Memory was assessed by total freezing time in the same environment 24 h and 28 h after fear conditioning, and in an intervening test session in a different context. As expected, fimbria-fornix lesion impaired fear memory and dramatically attenuated hippocampal theta power. Artificial theta stimulation produced continuous theta oscillations that were almost similar to endogenous theta rhythm in amplitude and frequency. However, contrary to our predictions, artificial theta stimulation impaired conditioned fear response in both sham and fimbria-fornix lesioned animals. These data suggest that restoration of theta oscillation per se is not sufficient to support memory encoding after fimbria-fornix lesion and that universal theta oscillation in the hippocampus with a fixed frequency may actually impair memory.     

Some regional anatomical relationships of TRH to 5-HT in rat limbic forebrain

It is now a recognized principle that various neuropeptides are neuronally co-localized with biogenic amine or aminoacid neurotransmitters. In the rat CNS it has previously been shown that TRH is co-localized with 5-HT (and also with substance P) in cell bodies of the posterior raphe that project to the spinal cord. Although TRH cell bodies are known to be widely distributed throughout the forebrain there is no other known co-localization with 5-HT. In this study we further specify the anatomical relationship of TRH with 5-HT by use of surgical and neurotoxic lesioning with reference to limbic forebrain regions wherein TRH is greatly increased following seizures. In groups of rats, the fimbria-fornix was lesioned alone, or combined with a lesion of the dorsal perforant path or the ventral perforant path. There was a sham lesioned control group. Additional groups were lesioned with 5, 7 dihydroxytryptamine, 100 g i.v.t., 45 min. after i.p. desipramine, 25 mg/kg. All rats were sacrificed three weeks after lesions. Indoleamines were determined by HPLC in left anterior cortex, left pyriform/olfactory cortex, left dorsal hippocampus and left ventral hippocampus. TRH was determined by specific RIA in the corresponding right brain regions. The modal n was 7 rats. The surgical lesions reduced 5-HT to below the detection limit in dorsal hippocampus in all three groups, and to 31–52% of control in all the ventral hippocampus groups. 5-HIAA was reduced to 19–37% of control in dorsal and to 30–51% of control in ventral hippocampus. TRH was reduced to 44–61% of control in dorsal hippocampus and to 48–53% of control in ventral hippocampus. As was repeatedly observed in our previous reports all TRH levels in ventral hippocampus were higher than in dorsal hippocampus. The 5, 7 dihydroxytryptamine treatment nearly eliminated the indoleamines from all the forebrain regions examined while TRH levels were unchanged. These results can be explained by our previous data showing that immunoreactive TRH is intrinsic and localized to the vicinity of both CA and dentate granule cells of the hippocampus, but about half of hippocampal TRH enters via fibers of the fimbria-fornix. The perforant path appears to contribute no TRH to hippocampus, but, results with the combined lesion groups showed some reduction of 5-HIAA in ventral hippocampus as is expected from the known perforant path contribution of 5-HT. Since the neurotoxic lesion had no effect on TRH, the 5-HT pathway through the fimbria-fornix is probably anatomically separate from a parallel TRH pathway there. This study shows that co-localization of TRH with 5-HT is very unlikely in four specific limbic forebrain regions.Special issue dedicated to Dr. Morris H. Aprison.     

RU486对早孕猕猴海马GR的影响

目的探讨早孕猕猴给予大剂量RU486 3天后海马糖皮质激素受体(GR)的表达变化.方法 15只早孕猕猴随机分为空白对照组、赋形剂组和RU486组.空白对照组不予任何处理,赋形剂组和RU486组分别鼻饲赋形剂和RU486 3天.应用单克隆抗体链菌素亲生物蛋白过氧化酶(SP)免疫组织化学方法观察海马GR的表达情况,并用电子计算机图象分析技术进行处理.结果 RU486组猕猴妊娠终止,该组的海马GR表达显著下降,与对照组的差异有显著性.空白对照组和赋形剂组猕猴妊娠没有终止,其海马GR表达无差异.结论 RU486可使早孕猕猴海马的GR表达下降,推测这可能是该药终止妊娠的中枢作用机理之一.     

电磁脉冲对大鼠学习和脑内神经递质的影响

探讨电磁脉冲（ＥＭＰ）对大鼠神经系统的效应。实验采用Ｗｉｓｔａｒ大鼠,ＥＭＰ辐照后不同时间用Ｙ－型迷宫测其学习能力,高效液相色谱法检测脑不同部位的神经递质含量。与假照射组（对照组）相比,照后三天内各测定组大鼠学习能力降低（Ｐ＜０．０５）,其中照射后第１天组的海马内５－羟色胺（５－ＨＴ）和多巴酸（ＤＯＰＡＣ）含量升高（Ｐ＜０．０５）,下丘脑多巴胺（Ｄｏｐａｍｉｎｅ）含量升高（Ｐ＜０．０５）,肾上腺素（Ａｄｒ）含量降低;照后２天组海马Ａｄｒ含量降低（Ｐ＜０．０５）,海马５－ＨＴ含量升高（Ｐ＜０．０５）;照后３天组海马内Ａｄｒ含量降低（Ｐ＜０．０５）。ＥＭＰ能够改变大鼠不同脑区神经递质的含量,降低大鼠学习能力     

抗体应答期间大鼠脑和胸腺中儿茶酚胺含量的变化

目的：探讨在抗体应答期间,脑和淋巴器官中儿茶酚胺（ＣＡｓ）含量的动态变化,籍以了解免疫状态对中枢和外周ＣＡｓ神经活动的影响。方法：用绵羊红细胞（ＳＲＢＣ）免疫大鼠,在免疫后第２￣７ｄ应用高效液相色谱－电化学检测法（ＨＰＬＣ－ＥＣＤ）测定大鼠下丘脑、海马、脑干和胸腺中云甲肾上腺素（ＮＡ）、肾上腺素（Ａ）、多巴胺（ＤＡ）和高香草酸（ＨＶＡ）的含量。结果：①下且脑和海马内ＮＡ在抗体应答期间升高,而胸腺中     

海马内生长抑素与γ－氨基丁酸在大鼠穿梭箱主动回避反应中的作用

本实验以大鼠穿梭箱主动回避反应（ＡＡＲ）的习得和消退为学习记忆的指标,研究了海马内生长抑素（ＳＳ）和γ－氨基丁酸（ＧＡＢＡ）在学习记忆中的作用。结果如下：（１）经训练而建立了ＡＡＲ的大鼠,其海马内ＳＳ较对照组显著增高,而海马内ＧＡＢＡ含量却明显降低;（２）海马内注入ＳＳ的耗竭剂半胱胺（Ｃｙｓ,２０ｇ／Ｌ）使大鼠ＡＡＲ的习得受到明显损害,ＡＡＲ的消退显著加速,海马内ＳＳ明显降低,而ＧＡＢＡ含量却显著升高;（３）海马内注入ＧＡＢＡ（２００ｇ／Ｌ）使大鼠ＡＡＲ的消退显著加速的同时,其海马内ＳＳ含量亦显著降低。由此表明,海马内ＳＳ可能有促进学习记忆的作用,而海马内ＧＡＢＡ升高则有相反的效应;二者在海马调控学习记忆过程中具有重要作用。     

急性低氧对血浆皮质醇浓度及海马去甲肾上腺素含量的影响

本工作观察不同氧浓度,不同低氧时间猫血浆皮质醇浓度和海马内去甲肾上腺素含量的改变,以分析海马在低氧应激过程中的作用,结果显示,吸入１０．４％及７．９％Ｏ２后,猫血浆皮质醇浓度逐渐升高,分别于４５ｍｉｎ及３０ｍｉｎ达峰值;而低氧３０ｍｉｎ后,海马组织中的ＮＥ含量明显降低,两者与低氧程度无明显关系。