鳜鱼消化道黏液细胞和6种酶的组织化学定位

采用阿利新蓝-过碘酸雪夫氏(AB-PAS)染色和酶组织化学方法对鳜鱼消化道各部位黏液细胞和6种酶的分布与定位进行了研究。结果显示,黏液细胞可为分为4种类型,食道黏液细胞多数为Ⅲ型和Ⅳ型,未见Ⅰ型和Ⅱ型;胃贲门和胃幽门黏膜上皮仅有Ⅰ型黏液细胞;胃体黏膜上皮则以Ⅲ型细胞为主;幽门盲囊中主要为Ⅱ型细胞;前肠和中肠中Ⅳ型黏液细胞最多,Ⅰ型最少;后肠黏液细胞则以Ⅳ型和Ⅱ型为主。酸性磷酸酶(ACP)主要分布于幽门盲囊和前肠的黏膜上皮;碱性磷酸酶(ALP)主要分布于食道、幽门盲囊和整个肠道黏膜上皮;非特异性酯酶(NSE)主要分布于胃幽门、中肠和后肠黏膜上皮;过氧化物酶(POX)在胃幽门黏膜上皮中活性较高;琥珀酸脱氢酶(SDH)主要分布于胃腺中;腺苷三磷酸酶(ATPase)在消化道各部位均有较多分布。鳜鱼消化道黏液细胞和酶的分布型与其它动物有相似之处,也有其一定的特异性,与消化道不同部位的消化吸收机能相适应。     

四指马鲅稚鱼、幼鱼和成鱼消化道黏液细胞组织化学研究

采用常规石蜡组织切片及阿利新兰–高碘酸雪夫–试剂(AB-PAS,AB,p H2.5)染色方法对四指马(Eleutheronema tetradactylum)稚鱼、幼鱼和成鱼消化道中黏液细胞的组织化学特征进行观察。35日龄稚鱼消化道黏液细胞数量少,种类单一,食道含数量较多的II型黏液细胞,偶见I型黏液细胞;幽门盲囊未检测出黏液细胞的分布;前肠以I型黏液细胞为主,有少量III型黏液细胞、中肠和后肠均只有II型黏液细胞。65日龄幼鱼消化道各段黏液细胞数量明显增多,种类丰富,食道、前肠和中肠均含I、II、III、IV型4种类型的黏液细胞,其余均含至少两种类型的黏液细胞,食道黏液细胞密度最大,其次是后肠。成鱼食道、幽门盲囊均含II型和IV型黏液细胞,但后者两型黏液细胞数量都比较少;胃部含有大量I型黏液细胞,胃腺区域则以IV型黏液细胞为主;肠道只含III、IV型黏液细胞,黏液细胞密度从前到后呈递增规律。稚鱼、幼鱼、成鱼消化道黏液细胞的分布特点体现消化道结构功能逐渐发育完善的规律。     

圆尾鲎消化道组织结构与黏液细胞分布

利用石蜡切片、苏木素-伊红(H.E)和阿利新兰-高碘酸雪夫试剂(AB-PAS,AB,pH 2.5)组化染色技术研究圆尾鲎(Carcinoscorpius rotundicauda)消化道组织结构及其黏液细胞的分布特征。圆尾鲎消化道外形为一直管状,中肠部位有分支。H.E染色结果显示,食道、胃、幽门、中肠与后肠的管壁一般结构由外至内分为外膜、肌肉层、黏膜下层、黏膜层。食道是一段短管状结构,肌肉层较胃壁的薄但几丁质层极厚。胃则为一膨大的砂囊结构,内含一定数量的纵行黏膜皱褶,肌肉纤维排列整齐,几丁质层较薄。幽门与中肠套叠,幽门壁肌肉层很薄,几丁质层清晰可见。中肠和后肠结构差异不大,具有一定数量的黏膜皱褶,上皮细胞间分布比较多的黏液细胞,均无几丁质层。AB-PAS组化染色结果显示,消化道有Ⅰ和Ⅱ型两种黏液细胞,不同部位分布数量差别很大。食道和幽门未见黏液细胞。胃黏膜下层有少量Ⅱ型黏液细胞。中肠和后肠黏液细胞数量比较多,尤其是与幽门套叠的中肠前端区域,均以Ⅱ型黏液细胞为主,主要分布在黏膜下层和黏膜上皮。在后肠黏膜下层有Ⅰ型黏液细胞分布,而黏膜上皮则分布密集的Ⅱ型黏液细胞。圆尾鲎消化道组织结构及黏液细胞分布特征反映其不同部位功能的差别,体现食性与消化机能相协调的特点。     

银鲳消化道形态与组织学结构特征及其消化酶活性

为了解银鲳(Pampus argenteus)消化道结构特点与其功能及食性的相关性, 采用解剖、石蜡切片、AB-PAS染色及酶活性检测技术对银鲳消化道的形态、组织结构、黏液细胞分布及消化酶活性进行研究。结果显示, 银鲳的消化道由口咽腔(舌)、食道侧囊、食道、胃及肠构成, 胃肠交界处有很多幽门盲囊。食道侧囊呈椭球形, 食道粗短, 胃呈U型, 肠有多个盘曲, 肠指数为2.03。舌上皮内有少量味蕾及较多黏液细胞。食道侧囊、食道、胃及肠均由黏膜层、黏膜下层、肌层及浆膜组成。食道侧囊内皱襞较发达, 被覆复层扁平上皮, 内含较多黏液细胞, 且以Ⅳ型为主, 皱襞顶端及侧面有内含角质刺的次级突起; 黏膜下层及肌层中有固定皱襞的骨质脚根; 侧囊内胃蛋白酶活性较高。食道内皱襞较高, 被覆复层扁平上皮, 内含较多黏液细胞, 且以Ⅳ型为主。胃内皱襞发达, 被覆单层柱状上皮, 未见黏液细胞分布; 胃腺发达, 胃内蛋白酶活性较高。肠道内褶襞多, 高度呈先下降后上升趋势, 黏液细胞密度前、中肠较高, 后肠较低, 且均以Ⅰ型为主; 肠道内胰蛋白酶、脂肪酶、淀粉酶及碱性磷酸酶活性较高。幽门盲囊组织结构与肠相似。银鲳的消化道结构特点、黏液细胞分布及消化酶活性与其功能及偏肉食的杂食性相适应。     

三种淡水鱼消化道黏液细胞的组织化学染色

目的：探索淡水鱼消化道黏液细胞的组织化学染色。方法：利用阿利新蓝和过碘酸雪夫试剂染色对鲶鱼（Silurus aso-tus）、鲢鱼（Hypophthalmichthys molirtix）、鲫鱼（Carassius auratus）消化道黏液细胞的类型与分布进行了观察,结果：AB-PAS（Alcian blue and periodic acid schiff’s reation）染色后动物消化管黏液细胞表现为4个类型：Ⅰ型玫瑰红色、Ⅱ型蓝绿色、Ⅲ型紫红色、Ⅳ型蓝紫色。除鲫鱼外动物食管始部黏液细胞多为圆形或椭圆形,食管中部黏液细胞主要是Ⅱ型和Ⅳ型杯状细胞。除鲫鱼动物胃体柱状黏膜上皮细胞与胃腺颈部细胞主要为Ⅰ型和Ⅲ型,黏液颗粒主要集中在细胞的核上区。链鱼、鲶鱼胃腺黏液细胞为Ⅰ型和Ⅲ型。链鱼、鲶鱼小肠的杯状细胞主要为Ⅲ型和Ⅳ型。鲫鱼小肠杯状细胞主要有Ⅱ型和Ⅳ型。这三种动物大肠杯状细胞主要为Ⅱ型和Ⅳ型。     

巴西彩龟与中华鳖消化道粘液细胞组织化学染色

利用阿利新蓝和过碘酸雪夫试剂染色对巴西彩龟、中华鳖消化道粘液细胞的类型与分布进行了观察,结果显示消化道粘液细胞表现为4种类型：Ⅰ型玫瑰红色;Ⅱ型蓝绿色;Ⅲ型紫红色;Ⅳ型蓝紫色。食道前部粘膜上皮为复层上皮,粘液细胞形态多样,Ⅰ、Ⅱ、Ⅲ、Ⅳ类型均有,食道中部与后部粘膜上皮为柱状上皮,其内有密集排列的杯状细胞,主要是Ⅱ型、Ⅲ型和Ⅳ型细胞。胃体柱状粘膜上皮细胞与胃腺颈部细胞主要为Ⅰ型和Ⅲ型,粘原颗粒主要集中在细胞的浅部。胃腺粘液细胞主要为Ⅰ型和Ⅲ型,着色较浅。小肠杯状细胞主要为Ⅲ型和Ⅳ型。大肠、泄殖腔杯状细胞主要为Ⅱ型和Ⅳ型。     

大鳞副泥鳅仔稚鱼消化道黏液细胞分布及发育

利用组织学切片及阿利新蓝-过碘酸雪夫(AB-PAS)组化染色技术对0~50日龄大鳞副泥鳅(Paramisgurnus dabryanus)消化道各段黏液细胞的发育与分布进行显微观察和研究。大鳞副泥鳅黏液细胞分为Ⅰ、Ⅱ、Ⅲ和Ⅳ4种类型。消化道黏液细胞最早出现在4日龄仔鱼的口咽腔和食道。10~15日龄口咽腔和食道黏液细胞数量快速增长,15~20日龄肠道各型黏液细胞数量显著增长,20日龄后消化道黏液细胞分布广泛。随着仔稚鱼发育,消化道各部黏液细胞主要以Ⅲ和Ⅳ型细胞为主。根据大鳞副泥鳅仔稚鱼消化道黏液细胞的发育特点,5~10日龄和15~20日龄为其消化道功能发育的两个敏感时期,20日龄后消化道功能逐渐发育完善。建议加强对5~20日龄仔稚鱼的日常饲养管理以提高苗种成活率。     

饥饿对高体革鯻消化道黏液细胞分布的影响

利用石蜡组织切片与AB-PAS染色技术研究饥饿0天(S0)、10天(S10)、20天(S20)对高体革消化道黏液细胞形态及分布的影响。结果发现,高体革饥饿20天后存活率为100%,食道含有Ⅱ、Ⅲ、Ⅳ型黏液细胞, S0组黏液细胞总密度为(2 839.13±261.52)个/mm2,饥饿不影响食道黏液细胞的种类,但S10组黏液细胞总密度减少[(1 546.00±70.02)个/mm2], S20组则表现为增加[(3 095.00±162.04)个/mm2]。S0、S10、S20三组胃部黏液细胞分布差异不大。S0和S10组幽门盲囊含Ⅰ和Ⅱ型黏液细胞, S20组则只有Ⅱ型黏液细胞,总密度先增加后减少,分别是(215.22±21.52)、(466.31±63.05)、(369.46±43.49)个/mm2。肠道只含Ⅱ型黏液细胞,饥饿使其密度逐渐增多,尤其是后肠,S0、S10、S20组密度分别是(1 683.50±219.64)、(2 068.71±65.38)、(4 622.50±85.60)个/mm2。不同部位黏液细胞对饥饿的响应存在差异,反映消化道不同部位结构功能的差异,饥饿显著影响消化道中Ⅱ型黏液细胞的分布,推测Ⅱ型黏液细胞分泌的变化有可能是高体革响应饥饿应激的敏感指标之一。     

中华蟾蜍与黑斑蛙消化道黏液细胞的观察

目的：探索两栖类动物消化道黏液细胞的类型与分布规律。方法：利用阿利新蓝与过碘酸雪夫试剂染色法对中华蟾蜍（Bufo bufo gargazizans）、黑斑蛙（Rana nigromaculata）消化道黏液细胞进行石蜡切片和染色。结果与结论：黏液细胞表现为4个类型：Ⅰ型玫瑰红色;Ⅱ型蓝绿色;Ⅲ型紫红色;Ⅳ型蓝紫色。两种动物食管黏膜上皮中黏液细胞主要是杯状细胞,其中中华蟾蜍的黏液细胞主要为Ⅱ型和Ⅳ型细胞,黑斑蛙的黏液细胞主要Ⅲ型细胞。胃体柱状黏膜上皮细胞与胃腺颈细胞主要为Ⅰ型和Ⅲ型细胞,黏原颗粒主要集中在细胞的核上区。胃腺浅部有成团分布并着色较浅的黏液细胞,其中黑斑蛙胃腺黏液细胞主要为Ⅰ型和Ⅲ型细胞,中华蟾蜍胃腺黏液细胞主要为Ⅲ型和Ⅳ型细胞。小肠杯状细胞主要为Ⅲ型和Ⅳ型细胞。大肠杯状细胞主要为Ⅱ型和Ⅳ型细胞。     

Diet composition of the seahorses, Hippocampus guttulatus Cuvier, 1829 and Hippocampus hippocampus (L., 1758) (Teleostei, Syngnathidae) in the Aegean Sea

The diet composition of the European seahorses, Hippocampus guttulatus and Hippocampus hippocampus was determined based on the analysis of 279 and 19 specimens, respectively, collected in the Aegean Sea. The diet of both species was mainly based on Crustacea, with Amphipoda, Anomura Decapoda and Mysidacea being the dominant prey categories. ANOSIM analyses, however, indicated statistically significant differences in the diet of the two species as well as differences in the diet composition of non-brooding males, brooding males and females within each species. In H. guttulatus , stomach fullness percentages and vacuity coefficient values indicated that female individuals seem to have a higher feeding activity in relation to males.     

Short- and long-term changes in extracellular glutamate and acetylcholine concentrations in the rat hippocampus following hypoxia

Hypoxia at birth is a major source of brain damage and it is associated with serious neurological sequelae in survivors. Alterations in the extracellular turnover of glutamate (Glu) and acetylcholine (ACh), two neurotransmitters that are essential for normal hippocampal function and learning and memory processes, may contribute to some of the neurological effects of perinatal hypoxia. We set out to determine the immediate and long-lasting effects of hypoxia on the turnover of these neurotransmitters by using microdialysis to measure the extracellular concentration of Glu and ACh in hippocampus, when hypoxia was induced in rats at postnatal day (PD) 7, and again at PD30. In PD7 rats, hypoxia induced an increase in extracellular Glu concentrations that lasted for up to 2.5 h and a decrease in extracellular ACh concentrations over this period. By contrast, perinatal hypoxia attenuated Glu release in asphyxiated rats, inducing a decrease in basal Glu levels when these animals reached PD30. Unlike Glu, the basal ACh levels in these animals were greater than in controls at PD30, although ACh release was stimulated less strongly than in control animals. These results provide the first evidence of the initial and long term consequences of the hypoxia on Glu and ACh turnover in the brain, demonstrating that hypoxia produces significant alterations in hippocampal neurochemistry and physiology.     

An Amphiphile-Dependent Form of Human Brain Caudate Nucleus Acetylcholinesterase: Purification and Properties

Abstract: Different forms of acetylcholinesterase (AChE), EC 3.1.1.7, were demonstrated in human brain caudate nucleus. One form was solubilized at high ionic strength, the other with Triton X-100. The detergent-extractable form was purified to homogeneity by affinity chromatography. This form of AChE is amphiphile-dependent; i.e., it was active only in the presence of amphiphiles (detergents or lipids). Further, the enzyme was shown to bind detergents and to interact hydrophobically with Phenyl-Sepharose. In the presence of detergents the enzyme is a tetramer (subunit molecular weight, 78,000) which aggregates on the removal of detergents. Human brain AChE showed a reaction of identity with human erythrocyte AChE in crossed-line immunoelectrophoresis. The high-salt-soluble brain enzyme did not cross-react with the erythrocyte enzyme. The two classes of AChE seem not to be related, as they show no common antigenic determinant.     

Extra-hippocampal projections of CCK neurons of the hippocampus and subiculum

A population of neurons in the hippocampus and subiculum contains cholecystokinin (CCK). Following transection of the dorsal fornix, a major afferent pathway of the hippocampus and associated structures. CCK levels were reduced in the septum and hypothalamus. A microdissection analysis indicated that the loss of CCK occurred in nuclei receiving direct projections from the hippocampus and subiculum, suggesting that CCK-containing neurons in the hippocampus and subiculum project to extrahippocampal regions.     

Introduction to What are the parietal and hippocampal contributions to spatial cognition?, the proceedings of a Discussion held at The Royal Society

Introduction to What are the parietal and hippocampal contributions to spatial cognition?, the proceedings of a Discussion held at The Royal Society on 19 and 20 March 1997. Organized by N. Burgess and J. O''Keefe and edited by N. Burgess, K. J. Jeffery and J. O''Keefe.
     

Balanced GABAergic and glutamatergic synapse development in hippocampal neurons

Coordinated development of excitatory and inhibitory synapses is crucial for normal function of neuronal circuits. Using homo- and heterochronic cultures of hippocampal neurons, we compared the formation of glutamatergic and GABAergic synapses at different stages and asked whether the age of dendrites affects their ability to accept new glutamatergic and GABAergic synapses. Neurons were transfected with either CFP-actin as a dendritic marker or GFP-synaptophysin as a presynaptic marker. We found that GFP-synaptophysin clusters formed on CFP-actin-labeled dendrites at similar density regardless of pre- and postsynaptic cell type or the age of dendrites (0-2 weeks) upon co-culturing. Therefore, the age of mature dendrites does not affect their ability to accept new synapses. Because GABAergic transmission switches from depolarizing to hyperpolarizing during 1-2 weeks in these cultures, our observations also suggest that this developmental switch does not alter the formation of GABAergic synapses.     

抑郁症大鼠海马神经元凋亡率和自噬相关蛋白的改变

目的 观察抑郁症模型大鼠海马神经元形态结构改变及凋亡、自噬的变化,探讨抑郁症海马体积异常的机制.方法 选用雄性成年SD大鼠,随机分为正常对照组和模型组,通过给予不可预见慢性温和应激建立抑郁症模型;采用尼氏染色、透射电镜技术观察海马神经元形态变化,流式细胞术检测海马神经元凋亡,采用透射电镜观察海马神经元自噬体,Western blott检测自噬相关蛋白LC-3和Beclin 1.结果 与对照组比较,模型组海马神经元体积萎缩,数量减少,细胞凋亡率增高（P〈0.05）.模型组海马神经元胞质内可见自噬体;与对照组相比,模型组LC-3Ⅱ蛋白和LC-3Ⅱ/LC-3Ⅰ比值增高,Beclin-1相对表达增高（P〈0.05）.结论 抑郁症大鼠海马神经元存在体积萎缩现象,可能与神经元凋亡和自噬增强有关.     

Regulation of Serotonin Release from the In Vitro Rat Hippocampus: Effects of Alterations in Levels of Depolarization and in Rates of Serotonin Metabolism

We analyze the time course of 5-hydroxytryptamine (5-HT, serotonin) release from K+-depolarized hippocampal slices using a two-compartment kinetic model. The model is based on the assumptions that the rate of release is dependent on the amount of 5-HT in a releasable pool and that this pool may be resupplied during depolarization by newly synthesized 5-HT. Comparisons were made between predictions of the model and observed changes in 5-HT metabolism and in 5-HT release studied under a variety of experimental conditions. In accordance with predictions of the model, experimental manipulation of 5-HT synthesis and breakdown rates did not affect release immediately after depolarization but did affect the release rate during prolonged depolarization. Increasing bath tryptophan from 0 to 10 microM approximately doubled both 5-HT synthesis and the release rate after 40 min of K+-induced depolarization while having a smaller effect on release during the first 2 min. Inhibition of 5-HT breakdown did not significantly affect release during the first 2 min of depolarization but increased it over threefold after 40 min. In contrast, altering the concentrations of K+ or Ca2+ in the incubation medium affected mainly the early phase of 5-HT release and not the late phase. Reducing Ca2+ from 2.4 to 0.4 mM reduced 5-HT release by about 30% during the first 9 min of depolarization but did not affect release during the subsequent 30 min. Increasing the concentration of K+ from 18 to 60 mM stimulated release by sixfold during the first 2 min but only twofold after a subsequent 30 min. These results support our kinetic model and suggest that regulation of 5-HT metabolism at the site of the nerve terminal could be a mechanism for modulation of 5-HT release during prolonged discharge of serotonergic neurons.     

Inositol Lipid Metabolism in Rat Hippocampal Formation Slices: Basal Metabolism and Effects of Cholinergic Agonists

Incubation of rat hippocampal formation slices under steady-state conditions with [3H]inositol leads to only three phospholipids becoming labelled: phosphatidylinositol, phosphatidylinositol 4-phosphate, and phosphatidylinositol 4,5-bisphosphate. All three lipids incorporate [32P]Pi into their phosphodiester phosphate group with the polyphosphoinositides also incorporating this tracer into their monoester phosphate groups. As the concentrations of these lipids remain constant during these labelling processes we conclude that the phosphodiester phosphate, the inositol moiety, and the monoester phosphate groups undergo metabolic turnover in hippocampal formation slices incubated in vitro. The rate of incorporation of [3H]inositol into all three inositol phospholipids was stimulated by the addition of methacholine to the medium. Moreover, following steady-state labelling of the inositol lipids with [3H]inositol, methacholine in the presence of 10 mM LiCl caused a transient fall of 13% in the radiochemical concentration of phosphatidylinositol 4,5-bisphosphate after only 30 s stimulation and a fall of 15% in the radiochemical concentration of phosphatidylinositol after 30 min. Concomitantly, there was an approximately stoichiometric rise in the radiochemical concentration of inositol phosphates. Thus, we suggest that methacholine stimulates an inositol phospholipid phosphoinositidase C in rat hippocampal formation slices.     

Occurrence of the Japanese seahorse Hippocampus mohnikei Bleeker 1854 from the Palk Bay coast of south-eastern India

The occurrence of the Japanese seahorse, Hippocampus mohnikei is reported for the first time from the Palk Bay coast of south-eastern India.