胆碱能和多巴胺能化合物对豚鼠“踏步自动作用”的影响

中枢M-胆碱受体阻滞剂东莨菪碱(0.2—0.5mg/kg)和多巴胺前体L-多巴[特别是″L-多巴 RO-4-4602″(300mg/kg 50 mg/kg)],能增强由4-氨基-2,2,5,5-四(三氟甲基)-氢化咪唑(6—8mg/kg)引起的豚鼠“踏步自动作用”。胆碱酯酶可逆性抑制剂毒扁豆碱(0.5mg/kg)和多巴胺受体阻断剂氟哌啶醇(0.1—0.2mg/kg)则减弱上述“踏步自动作用”,从而表明豚鼠的“踏步自动作用”不仅和胆碱能及多巴胺能系统的活动有关,而且可能和两者间的交互影响有关。此外,豚鼠规律的强踏步运动常见伴随θ节律的呈现。     

四氢小檗碱的中枢抑制作用与GABA受体无关

应用蚯蚓背肌收缩反应,证实四氢小檗碱（ＴＨＢ,１０－７－１０－４ｍｏｌ／Ｌ）不能影响ＧＡＢＡ和ＡＣｈ两种受体的功能,多巴胺受体阻滞剂氟哌啶醇（ＨＡＬ）亦不能影响它们。由于异烟肼（ＩＮＨ）和氨基硫脲（ＴＳＣ）抑制ＧＡＢＡ的生物合成,印防己毒素（ＰＴ）和毕枯枯灵（Ｂｉｃ）作用于ＧＡＢＡ－ＢＺ受体复合体,它们都使小鼠产生惊厥,该作用能被氨基氧乙酸（ＡＯＡＡ）和安定（ＢＺ）所拮抗,但ＴＨＢ和ＨＡＬ却不能。由此表明前述ＴＨＢ的中枢抑制作用与ＧＡＢＡ抑制无关。     

家兔第四脑室内给予几种递质对膈神经放电高频振荡的影响

于65只清醒麻痹、迷走神经切断的家兔,观察第四脑室内注射中枢递质和受体阻断剂对膈神经放电高频振荡频率的影响。去甲肾上腺素、5-羟色胺、多巴胺、乙酰胆碱或γ-氨基丁酸,可使高频振荡频率下降。酚妥拉明、氟哌啶醇或麦角酰二乙胺对高频振荡频率无明显影响,而肉桂硫胺、东莨菪碱或印防已毒素可使高频振荡频率加快。结果提示,内源性5-羟色胺、乙酰胆硷和γ-氨基丁酸可能参与呼吸的调节。     

家兔第四脑室注射乙酰胆碱对肺动脉血压的影响

本工作将乙酰胆碱(ACh)注入麻醉家兔第四脑室,观察其对肺动脉血压的影响。结果发现(1)脑室注射50—100μg ACh后,肺动脉压和颈动脉压均下降。与此同时心率也出现一过性减慢。(2)切断两侧颈部迷走神经,ACh不再使心率减慢,但其降低肺动脉压和颈动脉压的作用不受到任何影响。(3)预先由第四脑室注射阿托品,可阻断AGh引起的肺动脉降压反应和颈动脉降压反应。(4)第四脑室注射六甲双铵或酚妥拉明,均不能阻断这二个降压反应。(5)第四脑室注射心得安不能阻断ACh引起的肺动脉降压反应,但能阻断ACh降低颈动脉压的作用。 实验结果表明:脑中ACh水平升高可通过激活胆碱能M-受体引起肺动脉压和颈动脉压下降;在ACh引起的颈动脉降压反应的中枢环节中有肾上腺素能β-受体活动参与;而且ACh降低肺动脉压和颈动脉压的作用不是通过迷走神经实现的,可能是由于延髓交感缩血管中枢紧张性降低所造成的。     

微波辐射对小鼠脑内乙酰胆碱含量及胆碱乙酸转移酶活性的影响

用频率为2450MHz功率密度为10mW/Cm~2(WBASAR约11.4W/kg)的微波(连续波)对置于微波暗室内的昆明种雄性小鼠急性全身照射1小时后,立即按常规方法断头,取脑,制成样品,然后用放射免疫测定法测量小鼠脑内乙酰胆碱(ACh)含量及胆碱乙酰转移酶(ChAT)活性。结果表明:照射组的ACh含量为11.6±1.4pmol/mg(脑鲜重),ChAT活性为45.4±8.7pmolACh/min.mg(脑鲜重);而对照组的分别为16.0±2.1pmol/mg和61.0±13.8pmolACh/min.mg。证明微波照射后可引起动物脑内ACh水平和ChAT活性下降,提示微波辐射对中枢胆碱能系统确有不利影响。     

多巴胺参与P物质对胃肌电活动和胃运动的毒蕈碱抑制效应

我们以前的工作表明尾核注射Ｐ物质能抑制胃肌电快波和胃运动,该效应是通过胆碱能Ｍ受体介导的。本文观察到Ｐ物质的抑胃效应可分别被尾核注射Ｐ物质抗血清、Ｐ物质受体拮抗剂［Ａｒｇ＾６,Ｄ－Ｔｒｉｐ＾７,ＭｅＰｈｅ＾８］－ＳＰ６－１１以及多巴胺Ｄ２受体了民剂氟哌啶醇所消除;尾核注射多巴胺对胃肌电快波和胃运动也有抑制作用,且能被尾核注射氟哌啶醇以及阿托品所阻断。据此,我们高想尾核Ｐ物质可能是通过多巴胺Ｄ２受体     

微波辐射对小鼠脑内乙酰胆碱含量及胆碱乙酸转移酶活性的影响

用频率为2450MHz功率密度为10mW/Cm~2(WBASAR约11.4W/kg)的微波(连续波)对置于微波暗室内的昆明种雄性小鼠急性全身照射1小时后,立即按常规方法断头,取脑,制成样品,然后用放射免疫测定法测量小鼠脑内乙酰胆碱(ACh)含量及胆碱乙酰转移酶(ChAT)活性。结果表明:照射组的ACh含量为11.6±1.4pmol/mg(脑鲜重),ChAT活性为45.4±8.7pmolACh/min.mg(脑鲜重);而对照组的分别为16.0±2.1pmol/mg和61.0±13.8pmolACh/min.mg。证明微波照射后可引起动物脑内ACh水平和ChAT活性下降,提示微波辐射对中枢胆碱能系统确有不利影响。     

高频电刺激下丘脑后核对氟哌啶醇致大鼠运动不能 的影响

目的：利用氟哌啶醇致僵直大鼠模拟帕金森病（PD）的运动不能,通过高频电刺激下丘脑后核(PH),观察大鼠僵直和运动能 力的变化,从而探讨PH 在PD治疗中潜在的应用价值。方法：将成年雄性SD 大鼠随机分为PH 刺激组、假刺激组和对照组,对 PH 刺激组和假刺激组大鼠双侧PH 置入双极刺激电极,腹腔注射氟哌啶醇30 min 后,PH刺激组给予持续高频电刺激(130 Hz,60 us, 100 uA),分别利用爬杆实验和跑步机实验评价大鼠僵直程度和运动能力。结果：腹腔注射氟哌啶醇1.0 mg/kg 后,①大鼠呈僵 直状态,其潜伏期为167.88± 17.88 s, 给予双侧PH 高频电刺激后潜伏期显著缩短至77.5± 21.27 s（P<0.01）。②跑步机试验显示大 鼠跑动速度和跑动距离显著下降,分别为5.78± 0.90 cm/s 和8.06± 4.35 m（P<0.01）,给予双侧PH高频电刺激后显著提高跑动速 度和跑动距离,分别为12.72± 3.66 cm/s 和98.61± 96.75 m（P<0.01）。结论：腹腔注射氟哌啶醇可模拟帕金森病的僵直和运动不 能症状,双侧高频电刺激PH 可显著拮抗氟哌啶醇对大鼠僵直和运动不能的作用,提示PH 为DBS治疗帕金森病运动不能的有效 刺激靶点,为临床DBS 刺激PH 治疗PD 提供实验依据。     

甲基 - 苯基四氢吡啶对脑不对称小鼠纹状体内多巴胺和白介素含量的影响

为探讨甲基 - 苯基四氢吡啶 (MPTP) 注射后脑不对称小鼠纹状体内多巴胺降低程度,及纹状体内细胞因子水平变化, C57BL/6J 小鼠经过伸爪取食试验,筛选为反映脑不对称的左利鼠和右利鼠,并接受 25 mg/kg MPTP 腹腔注射连续 5 天,检测注射后的第 1 天,第 3 天和第 14 天纹状体内多巴胺及代谢物含量和细胞因子 IL-1 β、 IL-6 的动态水平 . 结果表明,无论在左利鼠还是右利鼠,纹状体内多巴胺含量在 MPTP 注射后每个检测时间点都显著降低,纹状体内 IL-1 β水平在第 1 天显著降低,纹状体内 IL-6 水平在 MPTP 注射后每个检测时间点也显著降低 . 实验结果同时表明,左利鼠和右利鼠 IL-1 β和 IL-6 的基础水平有显著不同 . MPTP 注射后,与右利小鼠相比,左利小鼠有较高的多巴胺翻转降低和较低的细胞因子表达,而且,纹状体内多巴胺水平与纹状体内 IL-6 水平呈正相关 . 这些结果提示, MPTP 诱导多巴胺丢失伴随着黑质纹状体系统内细胞因子水平的改变,而且,脑不对称有可能通过影响纹状体内细胞因子水平而进一步影响 MPTP 诱导的多巴胺降低的程度 .     

肾多巴胺受体对血量扩张和脑室高张盐溶液注射引起促钠排泄的作用

本工作观察麻醉兔用氟哌啶醇阻断肾多巴胺受体对血量扩张（ＶＥ）和脑室高张盐溶液注射（ＩＣＨＮａ）引起促钠排泄反应的影响。在ＶＥ实验中。对照组与氟哌啶醇组排销量增加峰值分别为６５．６±１５．０与１９．０±５．５μｍｏｌ／ｍｉｎ（Ｐ＜０．０２）;在ＩＣＨＮａ实验中,对照组与氟哌啶醇组排钠量增加峰值分别为２８．９±４．６与２９．０±５．８μｍｏｌ／ｍｉｎ（Ｐ＞０．０５）,在肾多巴胺受体完好兔实验中,ＶＥ＋ＩＣＨＮａｇ;起的促钠排泄大于ＶＥ（Ｐ＜０．０１）或ＩＣＨＮａ（Ｐ＜０．０１）引起的反应。肾多巴胺受体阻断也可显著减弱ＶＥ＋ＩＣＨＮａ引起促钠排泄反应。结果表明：肾多巴胺受体阻断显著减弱ＶＥ以及ＶＥ＋ＩＣＨＮａ引起促钠排泄,对ＩＣＨＮａ的反应无效;在正常兔ＶＥ＋ＩＣＨＮａ引起促钠排泄显著大于各单一刺激的反应。     

Differential Labeling of Depot and Active Acetylcholine Pools in Nondepolarized and Potassium-Depolarized Rat Brain Synaptosomes

Abstract: To test the hypothesis that a pool of newly synthesized acetylcholine (ACh) turns over independently of preformed ACh, compartmentation and K⁺ -evoked release of ACh were examined in perfused synaptosomal beds intermittently stimulated by 50 m M K⁺. In resting synaptosomes, endogenous and labeled ACh was distributed between synaptic vesicles and the cytoplasm in a dynamic equilibrium ratio of 4:6. In the absence of new ACh synthesis, five sequential K⁺ -depolarizations caused a decremental release of preformed labeled ACh totaling 30% of the initial transmitter store. Further depolarization evoked little additional release, despite the fact that 60% of the labeled ACh remained in these preparations. Release of the preformed [¹⁴C]ACh was unaltered while new ACh was being synthesized from exogenous [³H]choline. Since the evoked release of [³H]ACh was maintained while that of [¹⁴C]ACh was decreasing, the [³H]ACh/[¹⁴C]ACh ratio in perfusate increased with each successive depolarization. This ratio was six to ten times higher than the corresponding ratio in vesicles or cytoplasm. These results indicate that the newly synthesized ACh did not equilibrate with either the depot vesicular or cytoplasmic ACh pools prior to release.     

蜜蜂脑乙酰胆碱免疫反应阳性神经元的分布

通过免疫组织化学方法-PAP法,使用乙酰胆碱(ACh)抗体,研究了中华蜂(Apis sinensis)和意大利蜂(Apis mellifera L)脑中具有乙酰胆碱免疫阳性反应的神经元胞体的形态、分布及神经元类型.并和已知的在其他昆虫脑中用ACh的合成酶ChAT和其水解酶AChE的抗体免疫组化法所获得的结果进行了比较.     

Multiple binding states of muscarinic acetylcholine receptors in membranes from neuroblastoma X glioma hybrid cells

Membranes of neuron-like NG108-15 hybrid cells bind [³H]quinuclidinyl benzilate (QNB) with high affinity and specificity. Greater than 90% of total [³H]QNB binding is to sites having the pharmacological specificity of muscarinic acetylcholine receptors. Three significant features characterize the interaction of ligands with these sites: (1) Specific binding of [³H]QNB at equilibrium follows a simple adsorption isotherm with an apparent K_D of 1 × 10^?10 M; (2) Rates of [³H]QNB association and dissociation are biphasic and, as the binding reaction proceeds, the fraction of readily dissociable [³H]QNB decreases; (3) Competition against [³H]QNB for specific binding sites by antagonists gives a slope of 1 when analyzed on Hill plots, but competition for binding sites by agonists gives a slope of less than 1. A simple two-step model for activation is proposed to account for these features.     

Choline stimulates the synthesis and accumulation of acetate in a cholinergic neuroblastoma clone

The NS 20 mouse neuroblastoma clone was shown to synthesize acetylcholine from labelled glucose or acetate as precursor of the acetyl moiety of acetylcholine; in both cases, the synthesis was stimulated by the presence of exogenous choline. In addition, we report that acetate is accumulated in the NS 20 clone by a mechanism that is highly temperature-dependent and is stimulated by the presence of externally added choline.     

Inhibition by trimethyl-tin,probenecid and phenylglyoxal of depolarization-induced acetylcholine release in Torpedo synaptosomes

The effects of trimethyl-tin (anion-hydroxyde ionophore, inhibiting oxydative phosphorylation and H⁺-ATPase) probenecid (inhibitor of anion transport in neural cells) and phenylglyoxal (arginine-specific reagent, inhibiting chloride exchanges in erythrocytes) were examined in Torpedo synaptosomes prepared from electric organ. All drugs significantly reduced the stimulated release of acetylcholine triggered by depolarization of nerve endings with high-K⁺ and/or gramicidin D. In contrast, trimethyl-tin, probenecid and phenylglyoxal did not affect the ionophore A23187-induced release of acetylcholine from the synaptosomes. The inhibitory potency of the compound trimethyl-tin was found to be similar to that of probenecid and phenylglyoxal on depolarization-induced acetylcholine release. This leads us to suggest that a relationship exists between modification of anion distribution during depolarization and acetylcholine release process. Moreover, since the release of ACh by calcium-ionophore A23187 was unaffected by trimethyl-tin, probenecid or phenylglyoxal, such compounds may also have an action on voltage-dependent Ca²⁺ flux across presynaptic membrane.     

Muscarinic acetylcholine receptor regulation by accelerated rate of receptor loss

Sustained activation of muscarinic acetylcholine receptors on neuron-like NG108-15 hybrid cells reduces the number of [³H]quinuclidinyl benzilate binding sites per cell as much as 88%. The response occurs at concentrations of agonists commensurate with those needed to occupy receptors and inhibit adenylate cyclase. Decreases in steady-state receptor levels persist as long as activator remains present. Withdrawal of activator results in a slow increase in receptor levels that is blocked by cycloheximide. Activation shortens receptor half-life by a factor of nearly 4, indicating that regulation occurs at the level of receptor breakdown.     

Age-Related Alterations In Pre-Synaptic and Receptor-Mediated Cholinergic Functions In Rat Brain

Fractional [³H]acetylcholine (ACh) release and regulation of release process by muscarinic receptors were studied in corpus striatum of young and aged rat brains. [³H] Quinuclidinyl benzilate (QNB) binding and carbachol stimulated phosphoinositide turnover, on the other hand, were compared in striatal, hippocampal and cortical tissues. High potassium (10 mM)-induced fractional [³H]ACh release from striatal slices was reduced by aging. Although inhibition of acetylcholinesterase with eserine (20 M) significantly decreased stimulation-induced fractional [³H]ACh release in two groups of rats, this inhibition slightly lessened with aging. Incubation of striatal slices with muscarinic antagonists reversed eserine-induced inhibition in fractional [³H]ACh release with a similar order of potency (atropine = 4-DAMP > AF-DX 116 > pirenzepine) in young and aged rat striatum, but age-induced difference in stimulated ACh release was not abolish by muscarinic antagonists. These results suggested that fractional [³H]ACh release from striatum of both age groups is modulated mainly by M₃ muscarinic receptor subtype. Although both muscarinic receptor density and labeling of inositol lipids with [myo-³H]inositol decreased with aging, carbachol-stimulated [³H]myo inositol-1-fosfat (IP₁) accumulation was found similar in striatal, cortical and hippocampal slices.     

Characterization of Acetylcholine-induced Increases in Cytosolic Free Calcium Concentration in Individual Rat Pancreatic β-cells

The intracellular free Ca²⁺ ion concentration ([Ca²⁺]i) was measured using fura-2 microspec-trofluorimetry in individual rat pancreatic β-cells prepared by enzymatic digestion and fluorescence-activated cell sorting. The mean basal concentration of [Ca²⁺]i in β-cells in the presence of 4.4 mM glucose and 1.8 mM Ca²⁺ was 112±1.6 nM (n=207). The action of acetylcholine (ACh) was concentration-dependent, and raising the concentration resulted in [Ca²⁺]i spikes of increasing amplitude and duration in some, but not all of the β-cells. In addition, the β-cells demonstrated variable sensitivity to ACh. The increases in [Ca²⁺]i were rapid, transient and were blocked by atropine at 10^?6M. A brief exposure to 50 mM K⁺ resulted in a transient increase in [Ca²⁺]i similar to that induced by ACh, but resistant to atropine. A high concentration of ACh (100μL 10^?4M or 10^?3M) induced [Ca²⁺]i oscillations in 11 out of 57 β-cells in the presence of 4.4 mM glucose. Using calcium channel blockers and Ca²⁺ free medium, the source of the increase in [Ca²⁺]i was deduced to be from extracellular spaces. Changing the temperature from 22 to 37°C did not affect the action of ACh on [Ca²⁺]i. These data strongly suggest that ACh exerted a direct action on [Ca²⁺]i in normal rat pancreatic β-cells and support a role for Ca²⁺ as a second messenger in the action of ACh.