丁香酚对大鼠弓状核温度敏感神经元电活动的影响

本实验采用微电极细胞外记录技术,在大鼠弓状核记录到单一放电单位６８例,其中热敏神经元１１例,占１６．２％,冷敏神经元２５例,占３６．８％;温度不敏感神经元３２例,占５３％。本实验同时观察了皮下注射丁香酚（２０μｇ／１００ｇ）对温度敏感神经元放电活动的影响,其结果为１０例冷敏神经元的放电频率均减慢;４例热敏神经元的放电频率均增快;而１４例温度不敏感神经元的放电活动则无明显改变。以上结果表明弓状核内存在以冷敏神经元居多的温度敏感神经元,并探讨了丁香酚的降温作用与它能够抑制弓状核的冷敏神经元,激活热敏神经元的电活动有关,亦成为弓状核参与体温调节的有力佐证     

回转器旋转对体外培养的鸡胚神经元的影响

用回转器旋转研究重力改变对体外培养的鸡胚神经元的影响。结果发现在回转器里经过７－９小时的处理后,神经元的形态和行为发生明显改变。只有６．３％的神经元呈现两极型的运动形态,而对照组正常培养的细胞运动形态的神经元占３４．２％。某些细胞的突起出现异常,呈现扭曲的形状。在培养基质上的神经元伸出许多丝状突起,丝状突起的末端锚定在周围的细胞上或锚定在培养基质上。经过重力改变处理的细胞重新放在相差显微镜下观察,结果显示神经元的运动活性降低,许多细胞没有观察到明显运动的迹象。     

葛根素,生石膏配伍对致热原作用下猫POAH温敏神经元放电…

应用微电极细胞外记录技术,在３４只猫ＰＯＡＨ区记录了温敏神经元单位放电,研究中药葛根素和生石膏解热可能的中机制。致热原使４例热敏神经元放电频率减少;使１１例冷敏神经元放电频率增加,注射等量葛根素和生石膏能反转上述作用。致热原及二药对５例温度不敏感神经元放电无影响。结果显示,葛根素和生石膏是影响致热原作用下ＰＯＡＨ区温敏神经元的电活动而解热的。二者配伍使用对冷敏神经元放电频率的影响比单独使用作用强,     

一种适合膜片钳单通道记录的大鼠DRG神经元急性分离方法

目的：建立一种适合膜片钳单通道记录的脊髓背根神经节神经元急性分离方法。方法：用酶消化和机械分离相结合的方法急性分离大鼠DRG神经元。结果：用本方法分离的DRG细胞容易形成较高的封接电阻（〉5GΩ）,降低了噪音干扰,可记录到pA级的单通道电流。结论：本方法急性分离的DRG神经元适合单通道膜片钳实验研究。     

以软刻技术微图案化培养纹状体神经元的形态学研究

探讨以软刻技术微加工带正电荷的聚乙烯亚胺(polyethyleneimine, PEI)图案,对培养纹状体神经元黏附存活及突起生长的影响．以软刻技术(微接触印刷方法)微加工三种不同的黏附底物：层粘连蛋白(laminin, LN)、带正电荷的多聚赖氨酸(poly-L-lysine, PLL)和PEI．新生乳鼠纹状体神经元体外分离培养,观察不同黏附底物对纹状体神经元黏附、存活和突起生长状态的影响,观察神经元在不同黏附底物上形成网络图案的差异．结果表明,在PEI 和PLL表面生长的神经元数量明显大于LN组,PEI与PLL、LN相比能形成更为完整的神经元图案．带强正电荷的PEI有助于神经元在其表面形成相对完整的图案,是构建体外神经网络的一种良好界面材料．     

神经元标本的简易制片法

神经元是一种有突起的细胞,在切片标本中突起常被切断,把神经元由组织中分离出来,经过染色制成永久性标本,有助于了解神经元的整体形态。现介绍一种神经元标本的简易制片法。试剂 30％酒精水溶液;中性卡红染液:卡红     

神经营养因子诱导分化的神经元样PC12细胞分裂的研究

神经营养因子（nerve growth factor,NGF）诱导PC12细胞分化产生的神经元样细胞一直被认为属于分裂后的细胞,没有分裂能力。然而在本研究中,我们观察了一些已经发生分化的PC12细胞,这些细胞长有很长的神经突起,在形态上属于神经元样细胞。在这些细胞中,我们不仅检测到DNA合成,而且观察到这些细胞的分裂现象。更令人感兴趣的是,除了胞体发生分裂外,位于胞体分裂位置的突起也一分为二,分别分配给两个子细胞。这些结果说明,形态发生分化的神经元样PC12细胞仍有分裂能力。本研究首次报道神经元样PC12细胞及其突起能发生分裂。     

葛根素、生石膏配伍对致热原作用下猫POAH温敏神经元放电的影响

应用微电极细胞外记录技术,在３４只猫ＰＯＡＨ区记录了温敏神经元单位放电,研究中药葛根素和生石膏解热可能的中枢机制。致热原使１４例热敏神经元放电频率减少;使１１例冷敏神经元放电频率增加。注射等量葛根素和生石膏能反转上述作用。致热原及二药对５例温度不敏感神经元放电无影响。结果显示,葛根素和生石膏是影响致热原作用下ＰＯＡＨ区温敏神经元的电活动而解热的。二者配伍使用对冷敏神经元放电频率的影响比单独使用作用强,提示在中枢水平二者有协同作用。     

高效的多巴胺能神经元原代培养方法及其影响因素的研究

目的：建立一种简单高效的中脑多巴胺神经元细胞原代培养方法,并观察胰酶消化对中脑多巴胺能神经元突起生长的损伤作用。方法：以Nakai等经典神经元细胞原代培养方法为基础,通过使用低日龄胎鼠,初次培养液加入胎牛血清等步骤,促进中脑多巴胺能神经元细胞贴壁和生长;在无胰酶消化组直接使用内口外翻的小口径硅化吸管轻柔吹打离散细胞,比较两种方法间神经元细胞突起形成的差异。结果：接吹打组其多巴胺能神经元细胞突起的生长程度（2124-10um）明显高于胰酶消化组（113＋9μm）（P〈0．01）,而两组间多巴胺阳性细胞比例未见显著差异（P〉0．05）。结论：在中脑多巴胺能神经元细胞原代培养中,低日龄胎鼠及免胰酶消化离散细胞可减少神经元细胞损伤,有利于细胞突起的生长。     

孕期家装污染对子代大鼠脑组织结构和认知功能的影响

目的:建立孕期家装污染大鼠模型,并探讨其对孕鼠子代神经元形态和神经行为的影响.方法:将22只成年孕鼠随机分为对照组和染毒组,分别在孕早期(D1-D10)给予相应的处理.在生后不同时间观察新生大鼠大脑的发育并检测脑组织重量,同时观察新生大鼠皮层神经元的形态变化并检测学习记忆活动.结果:各组新生大鼠脑组织无畸形,且各组大鼠脑组织重量无显著变化,但与对照组相比染毒组皮层神经元的排列及形态欠规则.出生一月后染毒组皮层神经元突起的数量及长度均少于对照组.行为学检测观察到染毒组大鼠其子代大鼠的学习记忆能力明显低于对照组.结论:家装污染物可导致新生大鼠海马神经元的形态学改变并抑制神经元突起的发育与生长,对大鼠的学习记忆功能有明显的损害作用.     

隔区和第Ⅲ脑室注射精氨酸加压素对家兔视前区—下丘脑前部温敏神经元放电的影响

为了研究精氨酸加压素（ＡＶＰ）的抗热机理,本研究观察了家兔隔区和第Ⅲ脑室微量注射ＡＶＰ对视前区一下丘脑前部（ＰＯ－ＡＮ）温度敏感神经元放电的影响。结果如下：（１）隔区注射ＡｖＰ能使ＰＯ－ＡＨ热敏神经元放电明显增加,冷敏神经元放电明显减少。（２）第Ⅲ脑室注射ＡＶＰ只能使部分ＰＯ－ＡＨ热敏神经元放电增加,冷敏神经元放电减少;而另外一部分热敏神经元和冷敏神经元则出现相反的效应。实验结果表明,隔区注射ＡＶＰ的抗热作用,可能是由于注射到隔区的ＡＶＰ使该区的神经元活动发生改变,而影响了ＰＯ—ＡＨ温度敏感神经元的活动所致。也提示隔区在体温调节中可能起重要作用。     

Neuronal basis of Hammel's model for set-point thermoregulation.

In 1965, H. T. Hammel proposed a neuronal model to explain set-point thermoregulation. His model was based on a synaptic network encompassing four different types of hypothalamic neurons: i.e., warm-sensitive and temperature-insensitive neurons and heat loss and heat production effector neurons. Although some modifications to this model are suggested, recent electrophysiological and morphological studies support many of the model's major tenets. Hypothalamic warm-sensitive neurons integrate core and peripheral thermal information. These neurons sense changes in hypothalamic temperature, and they orient their dendrites medially and laterally to receive ascending afferent input from cutaneous thermoreceptors. Temperature-insensitive neurons have a different dendritic orientation and may provide constant reference signals, which are important in determining thermoregulatory set points. In Hammel's model, temperature-sensitive and -insensitive neurons send mutually antagonistic synaptic inputs to effector neurons controlling various thermoregulatory responses. The model predicts that warm-sensitive neurons synaptically excite heat loss effector neurons and inhibit heat production effector neurons. In recent studies, one counterpart of these effector neurons may be "excitatory postsynaptic potential-driven neurons," the activity of which is dependent on synaptic excitation from nearby cells. Excitatory postsynaptic potential-driven neurons have sparse dendrites that appear to be specifically oriented, either medially or laterally, presumably to receive selective synaptic input from a discrete source. Another counterpart of effector neurons may be "silent neurons," which have extensive dendritic branches that may receive synaptic excitation from remote sources. Because some silent neurons receive synaptic inhibition from nearby warm-sensitive neurons, Hammel's model would predict that they have a role in heat production or heat retention responses.     

Preoptic hypothalamic warming suppresses laryngeal dilator activity during sleep

Upper airway dilator activity during sleep appears to be diminished under conditions of enhanced sleep propensity, such as after sleep deprivation, leading to worsening of obstructive sleep apnea (OSA). Non-rapid eye movement (NREM) sleep propensity originates in sleep-active neurons of the preoptic area (POA) of the hypothalamus and is facilitated by activation of POA warm-sensitive neurons (WSNs). We hypothesized that activation of WSNs by local POA warming would inhibit activity of the posterior cricoarytenoid (PCA) muscle, an airway dilator, during NREM sleep. In chronically prepared unrestrained cats, the PCA exhibited inspiratory bursts in approximate synchrony with inspiratory diaphragmatic activity during waking, NREM, and REM. Integrated inspiratory PCA activity (IA), peak activity (PA), and the lead time (LT) of the onset of inspiratory activity in PCA relative to diaphragm were significantly reduced in NREM sleep and further reduced during REM sleep compared with waking. Mild bilateral local POA warming (0.5-1.2 degrees C) significantly reduced IA, PA, and LT during NREM sleep compared with a prewarming NREM baseline. In some animals, effects of POA warming on PCA activity were found during waking or REM. Because POA WSN activity is increased during spontaneous NREM sleep and regulates sleep propensity, we hypothesize that this activation contributes to reduction of airway dilator activity in patients with OSA.     

隔区注射AVP和AVP抗血清对家兔温敏神经元放电的影响

精氨酸加压素可能是一种内源性退热剂,其抗热作用的最敏感点位于大脑边缘系统的隔区。为了研究ＡＶＰ抗热的作用的机理,本文观察了隔区注射ＡＶＰ和ＡＶＰ抗血清对家兔视前区－下丘脑前部温度敏感神经元放电活动的影响。实验结果如下：１（１）隔区注射ＡＶＰ能使ＰＯ－ＡＨ热敏神经元放电明显增加,冷敏神经元放电明显减少;而隔区注＝谢人工脑脊液对热敏神经元和冷敏神经元的放电均无明显影响。（２）隔区注射ＡＶＰ抗血清后,Ｐ     

甲状腺素与甲巯咪唑对大鼠视前区与下丘脑前部温度敏感神经元的影响

本文观察了皮下注射甲状腺素和胃饲甲巯咪唑对大鼠视前区-下丘脑前部(PO/AH)温度敏感神经元的影响。PO/AH 存在温度敏感神经元(TSN)和温度不敏感神经元(TIN),在 TSN 中又可分为热敏神经元(WSN)和冷敏神经元(CSN)。在正常大鼠,TSN∶TIN=1.43∶1;WSN∶CSN=1.86∶1。以上比例可被甲状腺素和甲巯咪唑所改变:前者使之分别下降为1∶2.36和1.20∶1;后者使 TSN∶TIN 下降为1∶1.29,WSN∶CSN 上升为2.40∶1。各组神经元构成比有显著性差异,(X_2=9.64,P<0.05)。正常大鼠中所记录到的高频神经元(>15H_z)占32%(11/34),甲状腺素组仅有8%(3/37),甲巯咪唑组为9%(3/32)。实验中还发现甲状腺素组和甲巯咪唑组中 TSN 对热刺激的耐受性显著低于正常组。以上结果提示:甲状腺素或甲巯咪唑所致体温上升或下降可能与 PO/AH 神经元兴奋性及比例等改变有关;甲状腺素合成、释放和代谢的紊乱可以在下丘脑水平干扰体温调节过程;但这一作用的具体途径有待于进一步研究。     

Hypothalamic mechanisms in thermoregulation

Certain preoptic and rostral hypothalamic neurons are sensitive to changes in local preoptic temperature (Tpo). These neurons also receive much afferent input from peripheral thermoreceptors and control a variety of thermoregulatory responses. In thermode-implanted animals, preoptic warming increases the firing rate in warm-sensitive neurons and elicits heat loss responses such as panting and sweating. Preoptic cooling increases the firing rate in cold-sensitive neurons and elicits, first, heat retention responses (e.g., cutaneous vasoconstriction and thermoregulatory behavior), then heat production responses (e.g., shivering and nonshivering thermogenesis). It is likely that the preoptic thermosensitive neurons control these thermoregulatory responses because both respond similarly to changes in Tpo and skin temperature. Specifically, skin warming not only increases panting, skin blood flow, and the firing rate of warm-sensitive neurons, but also decreases the sensitivity of all these responses to Tpo changes. Skin cooling not only increases metabolic heat production, heat retention behavior, and the firing rate of cold-sensitive neurons, but also increases the hypothalamic thermosensitivity of all these responses. Low-firing warm-sensitive neurons receive little afferent input and are most sensitive to high Tpo. Many of these low-firing neurons probably serve in controlling heat loss responses. High-firing warm-sensitive neurons receive much excitatory afferent input and are usually sensitive only to low Tpo. These neurons probably exert their greatest influence on heat production responses, possibly by inhibiting and, thus, determining the thermosensitive characteristics of nearby cold-sensitive neurons.     

Response of trigeminal ganglion neurons to thermal stimulation of the beak in pigeons

Summary The activity of neurons from the trigeminal ganglion of pigeons have been recorded while cooling or warming the beak. Thermosensitive neurons were seldom compared with mechanosensitive units. From a total of 16 thermosensitive neurons, 13 were excited by cooling and 3 by warming. The impulse frequency strongly depended on temperature. At constant temperatures, constant firing rates were established. The static curves of cold-sensitive units showed, that with decreasing temperature a nearly linear rise in firing rate occurred between about 36 °C and 20 °C. One quantified, warm-sensitive neuron showed, with increasing temperature, a nearly linear rise between about 35 °C and 44 °C. Sudden cooling or warming caused no pronounced overshoot as in mammals. Six cold-sensitive neurons were totally inhibited by rewarming as were 2 warm-sensitive units by cooling. No comparable influence of temperature on the discharge rate of some slowly-adapting mechanoreceptors (pressure stimulus) was exhibited.Supported by the Deutsche Forschungsgemeinschaft (SFB Bionach).     

Effects of GABA-transaminase inhibitor Vigabatrin on thermoregulation in rats

Vigabatrin is a GABA derivative (gamma-vinyl GABA) which inhibits irreversibly the enzyme activity of GABA transaminase and thus increased indirectly brain GABA concentrations. We have used body temperature assay to examine the effects of Vigabatrin on thermoregulation in intact rats. In order to understand the mechanism of thermoregulatory action of Vigabatrin at cellular level, we have investigated its effect on individual warm-sensitive preoptic area/anterior hypothalamus (PO/AH) neurons in rat brain slice preparations. The results of the present study suggest that Vigabatrin produced dose-dependent hypothermia in rats and also increased temperature sensitivity of warm-sensitive PO/AH neurons.     

Neuromuscular electrophysiology of the filarial helminth Dipetalonema viteae

1. A body wall preparation is described which permits intracellular recording from the somatic muscle cells of the small filarial nematode, Dipetalonema viteae. Using this preparation, resting membrane potentials were measured and spontaneous muscle depolarizations described. 2. Stimulatory effects noted upon the addition of acetylcholine, or the cholinergic agonists suggest the hypothesis that acetylcholine is the excitatory neurotransmitter. However, in contrast with vertebrate tissues, the cholinergic antagonists, d-tubocurarine, hexamethonium and pentolinium do not inhibit somatic muscle activity of the worm. 3. GABA inhibited somatic muscle depolarizations, suggesting the possibility that it may serve as an inhibitory neurotransmitter. 4. The anthelmintic drug, levamisole, produced a depolarizing block. Effects of other pharmacological agents are described, discussed and compared with effects on vertebrate muscles.     

会阴部皮肤和盆腔内脏的伤害感受性信息在猫骶髓后连合核神经元的汇聚

在戊巴比妥钠麻醉的猫上,用玻璃微电极细胞外记录的方法,观察了躯体和内脏的伤害性刺激对骶髓后连合核神经元活动的影响。结果表明,所有接受盆神经内Ａδ纤维传入的神经元皆为特异性伤害感受或广动力范围神经元－它们可被包括会阴部皮肤的躯体感受野的机械性及强电刺激诱发。上述结果提示,Ａδ纤维可能在盆腔内脏的伤害感受性传递中起重要作用。