Acetylcholine and GABA mediate opposing actions on neuronal chloride channels in crayfish

A central principle of neural integration is that excitatory and inhibitory neurotransmitters effect the opening of distinct classes of membrane ionic channels and that integration consists of the summation of the opposing ionic currents on the postsynaptic membrane. In tangential cells of crayfish optic lobes, a hyperpolarizing, biphasic synaptic potential is produced by the concurrent action of acetylcholine and gamma aminobutyric acid (GABA). Acetylcholine hyperpolarizes the cell and increases chlorine conductance. GABA depolarizes the cell by closing some of the same chloride channels. Therefore, in this case integration is achieved by the antagonistic actions of two transmitters on the same ionic channel.     

Sequential interplay of nicotinic and GABAergic signaling guides neuronal development

GABA (gamma-aminobutyric acid), the major inhibitory transmitter in the brain, goes through a transitory phase of excitation during development. The excitatory phase promotes neuronal growth and integration into circuits. We show here that spontaneous nicotinic cholinergic activity is responsible for terminating GABAergic excitation and initiating inhibition. It does so by changing chloride transporter levels, shifting the driving force on GABA-induced currents. The timing of the transition is critical, because the two phases of GABAergic signaling provide contrasting developmental instructions. Synergistic with nicotinic excitation, GABAergic inhibition constrains neuronal morphology and innervation. The results reveal a multitiered activity-dependent strategy controlling neuronal development.     

A selective imidazobenzodiazepine antagonist of ethanol in the rat

Ethanol, at pharmacologically relevant concentrations of 20 to 100 mM, stimulates gamma-aminobutyric (GABA) receptor-mediated uptake of 36Cl-labeled chlorine into isolated brain vesicles. One drug that acts at GABA-benzodiazepine receptors, the imidazobenzodiazepine Ro15-4513, has been found to be a potent antagonist of ethanol-stimulated 36Cl- uptake into brain vesicles, but it fails to antagonize either pentobarbital- or muscimol-stimulated 36Cl- uptake. Pretreatment of rats with Ro15-4513 blocks the anticonflict activity of low doses of ethanol (but not pentobarbital) as well as the behavioral intoxication observed with higher doses of ethanol. The effects of Ro15-4513 in antagonizing ethanol-stimulated 36Cl- uptake and behavior are completely blocked by benzodiazepine receptor antagonists. However, other benzodiazepine receptor inverse agonists fail to antagonize the actions of ethanol in vitro or in vivo, suggesting a novel interaction of Ro15-4513 with the GABA receptor-coupled chloride ion channel complex. The identification of a selective benzodiazepine antagonist of ethanol-stimulated 36Cl- uptake in vitro that blocks the anxiolytic and intoxicating actions of ethanol suggests that many of the neuropharmacologic actions of ethanol may be mediated via central GABA receptors.     

GABA对小麦幼苗耐盐性的影响

以小麦为试验材料,研究了NaCl胁迫下外源GABA浓度对小麦发芽率、芽长、根长、鲜干重、相对电导率及其酶活性的影响.结果表明:与对照相比,GABA浸种促进了小麦幼苗芽的生长,对根的生长影响较小;在GABA浓度为0.5 mmol.L-1条件下,小麦幼苗的发芽率及其鲜干重有明显的提高;相对电导率的测定结果显示,不同浓度GABA对膜都有不同程度的保护作用,在GABA浓度为0.5 mmol.L-1时效果最显著;对SOD、POD和CAT这几种酶活性的测定结果显示,随着GABA浓度的增加其活性都有不同程度的增加;由此推断,外源GABA可以减轻盐胁迫对小麦幼苗造成的伤害,具有一定的生理保护作用.     

铁钙复合法处理高浓度含磷农药废水的研究

[目的]探索由生产氟磺胺草醚而产生的高浓度含磷废水的除磷方法。[方法]用氯化钙、氯化铁及铁钙复合法对农药厂生产氟磺胺草醚而产生的高浓度含磷废水进行处理。[结果]氯化钙处理法的最佳pH值为8,最佳投药量为5.33 g/L,去除率为99.45%,处理成本为10.80元/kg磷;氯化铁处理法的最佳pH值为7,最佳投药量为6.50 g/L,去除率为99.08%,处理成本为56.89元/kg磷;铁钙复合法处理的最佳pH值为8,氯化钙最佳投药量为5.33 g/L,氯化铁最佳投药量为60 mg/L,此时去除率达99.91%,处理成本为11.28元/kg磷;处理后的废水可达到《污水综合排放标准（GB 8978—1996）》1级排放标准。[结论]铁钙复合法对该种农药废水中高浓度无机磷具有良好的去除效果。     

Brain glutamate decarboxylase cloned in lambda gt-11: fusion protein produces gamma-aminobutyric acid

Glutamate decarboxylase (GAD; E.C. 4.1.1.15) converts glutamate to gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the vertebrate central nervous system. This report describes the isolation of a GAD complementary DNA clone by immunological screening of a lambda gt-11 brain complementary DNA expression library. The fusion protein produced by this clone catalyzes the conversion of glutamate to GABA and carbon dioxide, confirming its identity as GAD. Antibodies to beta-galactosidase remove GAD enzymatic activity from solution, showing that this activity is associated with the fusion protein. In immunoblotting experiments all three available antisera to GAD reacted with the fusion polypeptide and with two major polypeptides (molecular size, 60,000 and 66,000 daltons) in brain extracts.     

Leonurine protects ischemia-induced brain injury via modulating SOD,MDA and GABA levels

The present study was designed to investigate the protective effects of leonurine, a compound purified from Herba Leonuri that is active on ischemic rat behavior and cortical neurons, and explore the underlying mechanism. The general rat activity, cortical neuron morphology, superoxide dismutase (SOD), malondialdehyde (MDA), g-aminobutyric acid (GABA) and glutamate decarboxylase 67 (GAD67) levels were measured. We found leonurine significantly improve the general activity of rats in an open-field test, which was associated with attenuated neuronal damage induced by ischemia. Moreover, serum SOD activity was significantly greater, MDA level lower in the leonurine group as compared with ischemia group. In addition, GABA content in the cerebral cortex was significantly greater in high-dose leonurine group. Correspondingly, GAD67 protein level coincided with the GABA level. Taken together, our results demonstrated that leonurine attenuated brain injury during ischemia via antioxidative and anti-excitotoxicity effects by targeting GABA and leonurine might become a useful adjuvant neuroprotective agent.     

Steroid hormone metabolites are barbiturate-like modulators of the GABA receptor

Two metabolites of the steroid hormones progesterone and deoxycorticosterone, 3 alpha-hydroxy-5 alpha-dihydroprogesterone and 3 alpha, 5 alpha-tetrahydrodeoxycorticosterone, are potent barbiturate-like ligands of the gamma-aminobutyric acid (GABA) receptor-chloride ion channel complex. At concentrations between 10(-7) and 10(-5)M both steroids inhibited binding of the convulsant t-butylbicyclophosphorothionate to the GABA-receptor complex and increased the binding of the benzodiazepine flunitrazepam; they also stimulated chloride uptake (as measured by uptake of 36Cl-) into isolated brain vesicles, and potentiated the inhibitory actions of GABA in cultured rat hippocampal and spinal cord neurons. These data may explain the ability of certain steroid hormones to rapidly alter neuronal excitability and may provide a mechanism for the anesthetic and hypnotic actions of naturally occurring and synthetic anesthetic steroids.     

Subthalamic GAD gene therapy in a Parkinson's disease rat model

The motor abnormalities of Parkinson's disease (PD) are caused by alterations in basal ganglia network activity, including disinhibition of the subthalamic nucleus (STN), and excessive activity of the major output nuclei. Using adeno-associated viral vector-mediated somatic cell gene transfer, we expressed glutamic acid decarboxylase (GAD), the enzyme that catalyzes synthesis of the neurotransmitter GABA, in excitatory glutamatergic neurons of the STN in rats. The transduced neurons, when driven by electrical stimulation, produced mixed inhibitory responses associated with GABA release. This phenotypic shift resulted in strong neuroprotection of nigral dopamine neurons and rescue of the parkinsonian behavioral phenotype. This strategy suggests that there is plasticity between excitatory and inhibitory neurotransmission in the mammalian brain that could be exploited for therapeutic benefit.     

γ-氨基丁酸对小麦耐盐性的影响

[目的]探究盐胁迫条件下γ-氨基丁酸(GABA)对小麦中光合活性和抗氧化活性的影响,进而揭示GABA提高小麦耐盐性的作用机理.[方法]测定不同浓度梯度GABA处理盐胁迫下小麦萌发幼苗的各生理参数,包括气体交换、光合作用量子产量、光合色素、抗氧化酶活性、丙二醛含量和相对电解质渗漏等,并分析比较不同处理对小麦苗光合活性和抗氧化活性的影响.[结果]外源GABA处理能提高小麦幼苗的光合活性和抗氧化活性,经0.75 mmol/L GABA处理后,小麦幼苗的光合活性和抗氧化活性显著提高(P<0.05),同时小麦生物膜的损伤得到修复.[结论]GABA是通过增强小麦的光合活性和抗氧化活性来提高小麦对盐胁迫的耐受性.     

GABAA-receptor function in hippocampal cells is maintained by phosphorylation factors

Gamma aminobutyric acid (GABA) mediates fast synaptic inhibition in the central nervous system by activating the chloride-permeable GABAA channel. The GABAA conductance progressively diminishes with time when the intracellular contents of hippocampal neurons are perfused with a minimal intracellular medium. This "run down" of the GABA-activated conductance can be prevented by the inclusion of magnesium adenosine triphosphate and calcium buffer in the intracellular medium. The amount of chloride conductance that can be activated by GABA is determined by competition between a calcium-dependent process that reduces the conductance and a phosphorylation process that maintains the conductance.     

Regulation of chloride channels by protein kinase C in normal and cystic fibrosis airway epithelia

Apical membrane chloride channels control chloride secretion by airway epithelial cells. Defective regulation of these channels is a prominent characteristic of cystic fibrosis. In normal intact cells, activation of protein kinase C (PKC) by phorbol ester either stimulated or inhibited chloride secretion, depending on the physiological status of the cell. In cell-free membrane patches, PKC also had a dual effect: at a high calcium concentration, PKC inactivated chloride channels; at a low calcium concentration, PKC activated chloride channels. In cystic fibrosis cells, PKC-dependent channel inactivation was normal, but activation was defective. Thus it appears that PKC phosphorylates and regulates two different sites on the channel or on an associated membrane protein, one of which is defective in cystic fibrosis.     

低频磁场对紫色红曲菌固态发酵产γ-氨基丁酸的影响

采用低频磁场处理紫色红曲菌(Monascus purpurcus),发酵产生γ-氨基丁酸(GABA)。分别研究了低频磁场强度、处理时间和处理时期对紫红曲霉固态发酵产GABA的影响,并研究了最佳处理条件下,紫红曲霉产GABA的发酵动力学。结果表明:用1.6 m T的低频磁场在紫色红曲菌发酵培养的第4~8 d,其产GABA的增长率提高了31.6%,最终产量增加了35.7%。低频磁场增强或抑制GABA产量可能与低频磁场改变其代谢途径有关。     

Interaction of convulsive ligands with benzodiazepine receptors

The gamma-aminobutyric acid (GABA)-benzodiazepine receptor complex, which is composed of distinct proteins embedded in the neuronal plasma membrane, is important for several effects of benzodiazepines, including protection afforded against convulsions. During structural modification of ethyl beta-carboline-3-carboxylate an agent was discovered which has high affinity for brain benzodiazepine receptors but which is a potent convulsant. Also in contrast to benzodiazepines, this type of benzodiazepine receptor ligand favors benzodiazepine receptors in the non-GABA-stimulated conformation, which may explain the convulsive properties.     

Altered regulation of airway epithelial cell chloride channels in cystic fibrosis

In many epithelial cells the chloride conductance of the apical membrane increases during the stimulation of electrolyte secretion. Single-channel recordings from human airway epithelial cells showed that beta-adrenergic stimulation evoked apical membrane chloride channel activity, but this response was absent in cells from patients with cystic fibrosis (CF). However, when membrane patches were excised from CF cells into media containing sufficient free calcium (approximately 180 nanomolar), chloride channels were activated. The chloride channels of CF cells were similar to those of normal cells as judged by their current-voltage relations, ion selectivity, and kinetic behavior. These findings demonstrate the presence of chloride channels in the apical membranes of CF airway cells. Their regulation by calcium appears to be intact, but cyclic adenosine monophosphate (cAMP)-dependent control of their activity is defective.     

硅烷偶联剂对桉木单板——聚氯乙烯膜复合材料性能的影响

为研究硅烷偶联剂对复合材料的性能影响,采用不同质量分数的硅烷偶联剂对桉木单板进行表面处理,然后与聚氯乙烯膜采用热压--冷压工艺制备木塑复合材料,测定复合材料的物理力学性能,并用扫描电子显微镜观察分析其界面相容机理。结果表明:当偶联剂质量分数为1%时处理效果最好,复合材料的胶合强度最高、耐水性能最好;当偶联剂质量分数为3%时,复合材料的弹性模量和静曲强度最高。单板经过硅烷偶联剂处理后,制得的复合材料的界面相容性得到改善。      

Southern corn leaf blight: susceptible and resistant mitochondria

Mitochondria isolated from etiolated shoots of blight-susceptible and blight-resistant corn plants were subjected, in various respiratory states, to the pathotoxin released by Helminthosporium maydis (race T). The addition of the pathotoxin to susceptible mitochondria caused respiratory rate and oxidative phosphorylation changes. The addition of pathotoxin to susceptible mitochondria suspended in a potassium chloride reaction medium induced an immediate and irreversible swelling regardless of the respiratory state of the mitochondria. This membrane swelling correlates with the observed respiratory and coupling effects of the pathotoxin. In all instances, mitochondria isolated from blightresistant corn failed to exhibit any of the above responses to the pathotoxin.     

小陷胸汤对鼠镇静安神作用的实验研究

目的观察小陷胸汤对小鼠及大鼠血液中、脑组织中5-HT(5-羟色胺)、GABA(r-氨基丁酸)镇静安神作用的影响。方法将50只KM种小鼠随机分为空白组、地西泮组、小陷胸汤高、中、低剂量组,灌胃7 d,观察各组小鼠的睡眠时间;将32只大鼠随机分为空白组、地西泮组、小陷胸汤组、小陷胸汤加地西泮组,于造模第8天开始,各组分别以不同药物连续灌胃8 d后,测定血液和脑组织中5-HT和GABA浓度。结果对小陷胸汤小鼠中剂量组与空白组平均睡眠持续时间比较差异有统计学意义(P<0.05)。小陷胸汤组大鼠与空白组血液和脑组织中GABA及5-HT浓度比较差异有统计学意义(P<0.05)。结论小陷胸汤能够对小鼠和大鼠产生镇静安神作用,其作用机制可能与提高5-HT、GABA的浓度有关。     

Cloning and expression of a rat brain GABA transporter

A complementary DNA clone (designated GAT-1) encoding a transporter for the neurotransmitter gamma-aminobutyric acid (GABA) has been isolated from rat brain, and its functional properties have been examined in Xenopus oocytes. Oocytes injected with GAT-1 synthetic messenger RNA accumulated [3H]GABA to levels above control values. The transporter encoded by GAT-1 has a high affinity for GABA, is sodium-and chloride-dependent, and is pharmacologically similar to neuronal GABA transporters. The GAT-1 protein shares antigenic determinants with a native rat brain GABA transporter. The nucleotide sequence of GAT-1 predicts a protein of 599 amino acids with a molecular weight of 67 kilodaltons. Hydropathy analysis of the deduced protein suggests multiple transmembrane regions, a feature shared by several cloned transporters; however, database searches indicate that GAT-1 is not homologous to any previously identified proteins. Therefore, GAT-1 appears to be a member of a previously uncharacterized family of transport molecules.     

Glucose, sulfonylureas, and neurotransmitter release: role of ATP-sensitive K+ channels

Sulfonylurea-sensitive adenosine triphosphate (ATP)-regulated potassium (KATP) channels are present in brain cells and play a role in neurosecretion at nerve terminals. KATP channels in substantia nigra, a brain region that shows high sulfonylurea binding, are inactivated by high glucose concentrations and by antidiabetic sulfonylureas and are activated by ATP depletion and anoxia. KATP channel inhibition leads to activation of gamma-aminobutyric acid (GABA) release, whereas KATP channel activation leads to inhibition of GABA release. These channels may be involved in the response of the brain to hyper- and hypoglycemia (in diabetes) and ischemia or anoxia.