Analysis of synaptic integration using the laser photoinactivation technique

Summary Crickets (and many other insects) have two antenna-like appendages at the rear of their abdomen, each of which is covered with hundreds of filiform hairs resembling the bristles on a bottle brush. Deflection of these filiform hairs by wind currents activates mechanosensory neurons at the base of the hairs. The axons from these sensory neurons project into the terminal abdominal ganglion to form a topographic representation (or map) containing information about the direction, velocity and acceleration of wind currents around the animal. Information is extracted from this map by primary sensory interneurons that are also located within the terminal abdominal ganglion. In this paper, we review the progress that has been made toward understanding the mechanisms underlying directional sensitivity of an identified sensory interneuron in the cricket,Acheta domesticus. The response properties of the cell have been found to depend to a large extent upon the structure of its dendritic branches, which determines its synaptic connectivity with the sensory afferents in the map of wind space and the relative efficacy of its different synaptic inputs.     

Switching among functional states by means of neuromodulators in the lobster stomatogastric ganglion

Summary The lobster stomatogastric ganglion contains the central pattern generators (CPGs) for the pyloric and gastric mill rhythms. All of the neurons and their synaptic connections have been identified for each rhythm and serve as the basis for understanding the mechanisms by which chemical neuromodulators are able to alter the functional state of each CPG. Using examples of different amines and peptides, I show how these substances can be found within specific neurons and how their application to the CPG can alter the motor patterns in specific ways. I also discuss what changes in cellular and synaptic properties occur as a result of bath application and particularly in the case of proctolin, how these changes may have behavioral correlates. The various outputs appear to be the result of a functional rewiring of anatomically defined neural circuits and this may be a widespread mechanism for the production of closely-linked but behaviorally distinct movement patterns.     

Can the binding of GABA,glycine and β-alanine to synaptic receptors be determined in the presence of a physiological concentration of Na+?

Summary Bicuculline- and strychnine-sensitive components of the binding of GABA, glycine and -alanine, which can be demonstrated in the presence of a physiological concentration of Na⁺, might be related to synaptic receptors.     

Memory processes during sleep: beyond the standard consolidation theory

Two-step theories of memory formation suggest that an initial encoding stage, during which transient neural assemblies are formed in the hippocampus, is followed by a second step called consolidation, which involves re-processing of activity patterns and is associated with an increasing involvement of the neocortex. Several studies in human subjects as well as in animals suggest that memory consolidation occurs predominantly during sleep (standard consolidation model). Alternatively, it has been suggested that consolidation may occur during waking state as well and that the role of sleep is rather to restore encoding capabilities of synaptic connections (synaptic downscaling theory). Here, we review the experimental evidence favoring and challenging these two views and suggest an integrative model of memory consolidation.     

Modification of glutamate receptors by phospholipase A2: its role in adaptive neural plasticity

Long-term potentiation (LTP) and long-term depression (LTD) are two electrophysiological models that have been studied extensively in recent years as they may represent basic mechanisms in many neuronal networks to store certain types of information. In several brain regions, it has been shown that these two forms of synaptic plasticity require sufficient dendritic depolarization, with the amplitude of the calcium signal being crucial for the generation of either LTP or LTD. The rise in calcium concentration mediated by the N-methyl-D-aspartate (NMDA) subtype of glutamate receptors has been proposed to stimulate various calcium-dependent enzymatic processes that could convert the induction signal into long-lasting changes in synaptic structure; protein kinases and phosphatases have so far been considered predominantly with regard to LTP and LTD formation. According to several lines of experimental evidence, changes in synaptic function observed with LTP and LTD are thought to be the result of modifications of postsynaptic currents mediated by the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) subtype of glutamate receptors. Moreover, it has become apparent recently that activation of the calcium-dependent enzyme phospholipase A2 (PLA2) could be part of the molecular mechanisms involved in alterations of AMPA receptor properties during long-term changes in synaptic operation. In the present review, we will first describe the results that indicate a critical role of the phospholipases in regulating synaptic function. Next, sections will be devoted to the effects of PLA2 and phospholipids on the binding properties of glutamate receptors, and a revised biochemical model will be presented as an attempt to integrate the PLA2 enzyme into the mechanisms ( in particular kinases and phosphatases) that participate in adaptive neural plasticity. Finally, we will review data relevant to the issue of selective changes in AMPA binding after environmental enrichment and LTP.     

Excitatory effect of the superior colliculus on the motor neurons of extra-ocular muscles of cats]

The synaptic actions which might underline the caccadic eye movements induced by stimulation of the superior colliculus have been studied in the unanesthetized cat "encéphale isolé". Intracellular recordings of extraocular muscle medial rectus motoneurons have revealed a minimally disynaptic excitatory projection from the superior colliculus to the ipsilateral medial rectus motoneurons (latency 1.3 to 2.6 msec).     

Comparative cytochemical and pharmacological studies on the cholinergic innervation of the branchial heart of the cephalopodSepia officinalis (L.)

Summary The dense motor innervation of the obliquely striated muscle cells in the branchial heart of Coleoida is composed of activating and inhibiting parts. The inhibiting cholinergic system investigated in this study is characterized histo- and cytochemically by a high acetylcholinesterase activity (EC 3.1.1.7) in the glycocalices of the nerve fibers with transparent synaptic vesicles, the muscle cells and, to a lesser extent, the ovoid interstitial cells, the functions of which are endocytosis and storage of catabolic substances. The pharmacological results from the isolated organ indicate a more nicotinic type of Ach-receptor, which can be reversibly blocked by D-tubocurarine and -bungarotoxin, but not with the same intensity by tetraethylammonium or atropine.     

慢性酒精中毒小鼠小脑的超微结构实验研究

目的 探讨慢性酒精中毒导致神经系统损伤的机制.方法 建立小鼠慢性酒精中毒动物模型,观察动物行为学的改变,测量血浆酒精浓度,通过透射电镜了解小脑的超微结构变化.结果 酒精处理组的血浆酒精浓度为101.4±20.5 mg/dL,与对照组和配对对照组比较,酒精处理组小鼠行动欠灵活,小脑线粒体形状多样、大小多变、数量增加和平均横断面面积显著减小;突触的数量减少、突触后膜致密物质厚度变薄、突触活性区长度变短及突触间隙宽度变宽,突触前结构内附着于突触的囊泡较多.结论 酒精对线粒体和突触结构、功能的损害可能是慢性酒精中毒的神经系统损伤机制之一.     

Genetic and molecular analysis of the synaptotagmin family

Secretion is a fundamental cellular process used by all eukaryotes to insert proteins into the plasma membrane and transport signaling molecules and intravesicular proteins into the extracellular space. Secretion requires the fusion of two phospholipid bilayers within the cell, an energetically unfavorable process. A conserved repertoire of vesicle-trafficking proteins has evolved that function to overcome this energy barrier and temporally and spatially control membrane fusion within the cell. Within neurons, opening of synaptic calcium channels and subsequent calcium entry triggers synchronous synaptic vesicle exocytosis and neurotransmitter release into the synaptic cleft. After fusion, synaptic vesicles undergo endocytosis, are refilled with neurotransmitter, and return to the vesicle pool for further rounds of cycling. It is within this local synaptic trafficking pathway that the synaptotagmin family of calcium-binding synaptic vesicle proteins has been postulated to function. Here we review the current literature on the function of the synaptotagmin family and discuss the implications for synaptic transmission and membrane trafficking. Received 14 August 2000; received after revision 20 September 2000, accepted 14 October 2000     

Synaptic vesicles and other organelles of human mossy fibre endings. A morphometric study

Summary The mossy fibre ending organelles of 6 men were studied, using morphometric methods. The numerical densities of agranular, coated and dense core vesicles, as well as the number of agranular vesicles per m² of the synaptic surface and per synapse were calculated.This work was granted by INIC: Centro de Morfologia Experimental (MbP1) e Centro de Anatomia Patológica e Oncologia (MbP3).     

'Soup' vs. 'sparks': Alexander Forbes and the synaptic transmission controversy

During the twentieth century, a controversy raged over the role of electrical forces and chemical substances in synaptic transmission. Although the story of the 'main' participants is well documented, the story of 'lesser' known participants is seldom told. For example, Alexander Forbes, who was a prominent member of the axonologists, played an active role in the controversy and yet is seldom mentioned in standard accounts of the controversy. During the 1930s, Forbes incorporated chemical substances into his theory of synaptic transmission, advocating a complementarity model for the role of electrical forces and chemical substances. By focusing on Forbes and the axonologists, the controversy is simply more than a debate over 'soup' vs. 'sparks' but also involves the relative roles of electrical forces and chemical substances in synaptic transmission. The implications of this case study for the nature of scientific controversies are also discussed.     

抑郁模型大鼠海马突触的差异蛋白质组学分析

目的通过抑郁模型大鼠海马突触的差异蛋白质组学分析,为抑郁症发病机制研究提供依据。方法根据糖水消耗基线值将40只健康雄性Sprague—Dawley大鼠随机分为慢性轻度不可预见性应激组和对照组（每组20只）。通过CUMS实验模式建立大鼠抑郁模型后,运用蔗糖密度梯度离心法提取两组大鼠的海马突触并用透射电镜进行检测。采用传统的双向凝胶电泳和基质辅助激光解析电离飞行时间串联质谱进行差异蛋白质组学分析。结果在行为学评价中CUMS组大鼠糖水消耗量和偏好度均降低（P〈0．01）,表明大鼠抑郁模型建立成功。通过结合双向凝胶电泳和质谱分析,共得到6个在CUMS组表达上调和10个在CUMS组表达下调的蛋白质,这些蛋白质的功能主要归为四类,即囊泡调节／递质释放／信号转导、能量代谢、细胞骨架、物质代谢。结论通过对抑郁模型大鼠海马突触的比较蛋白质组学分析及后续的蛋白质功能预测,发现了一些与突触传导功能障碍可能相关的蛋白质,从而为抑郁症发病机制的研究提供了有价值的线索。     

The molecular machinery of synaptic vesicle exocytosis

At the synapse, neurotransmitters are released via Ca(2+)-triggered exocytotic fusion of synaptic vesicles with the presynaptic plasma membrane. Synaptic vesicle exocytosis seems to share many basic principles and homologous proteins with other membrane fusion events. Conserved components of the general fusion machinery that participate in synaptic vesicle exocytosis include soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), ATPase N-ethylmaleimide-sensitive factor, Munc18/nSec1, Rab3 GTPase, and the exocyst proteins. In addition, synaptic vesicle exocytosis uses a set of unique components, such as synaptotagmin, complexin, Munc13, and RIM, to meet the special needs of fast Ca(2+)-triggered neurotransmitter release. This review summarizes present knowledge about the molecular mechanisms by which these components mediate and/or regulate synaptic vesicle exocytosis.     

Characterization of neurite outgrowth and ectopic synaptogenesis in response to photoreceptor dysfunction

In the mammalian retina, light signals generated in photoreceptors are passed to bipolar and horizontal cells via synaptic contacts. In various pathological conditions, these second-order neurons extend neurites into the outer nuclear layer (ONL). However, the molecular events associated with this neurite outgrowth are not known. Here, we characterized the morphological synaptic changes in the CNGA3/CNGB1 double-knockout (A3B1) mouse, a model of retinitis pigmentosa. In these mice, horizontal cells looked normal until postnatal day (p) 11, but started growing neurites into the ONL 1 day later. At p28, the number of sprouting processes decreased, but the remaining sprouts developed synapse-like contacts at rod cell bodies, with an ultrastructural appearance reminiscent of ribbon synapses. Hence, neurite outgrowth and ectopic synaptogenesis in the A3B1 retina were precisely timed events starting at p12 and p28, respectively. We therefore performed microarray analysis of retinal gene expression in A3B1 and wild-type mice at those ages to evaluate the genomic response underlying these two events. This analysis identified 163 differentially regulated genes in the A3B1 retina related to neurite outgrowth or plasticity of synapses. The global changes in gene expression in the A3B1 retina were consistent with activation of signaling pathways related to Tp53, Smad, and Stat3. Moreover, key molecules of these signaling pathways could be localized at or in close proximity to outgrowing neurites. We therefore propose that Tp53, Smad, and Stat3 signaling pathways contribute to the synaptic plasticity in the A3B1 retina.     

‘Soup’ vs. ‘Sparks’: Alexander Forbes and the Synaptic Transmission Controversy

During the twentieth century, a controversy raged over the role of electrical forces and chemical substances in synaptic transmission. Although the story of the ‘main’ participants is well documented, the story of ‘lesser’ known participants is seldom told. For example, Alexander Forbes, who was a prominent member of the axonologists, played an active role in the controversy and yet is seldom mentioned in standard accounts of the controversy. During the 1930s, Forbes incorporated chemical substances into his theory of synaptic transmission, advocating a complementarity model for the role of electrical forces and chemical substances. By focusing on Forbes and the axonologists, the controversy is simply more than a debate over ‘soup’ vs. ‘sparks’ but also involves the relative roles of electrical forces and chemical substances in synaptic transmission. The implications of this case study for the nature of scientific controversies are also discussed.     

Pentobarbital: Presynaptic effect in the squid giant synapse

Summary In concentrations that produced synaptic blockade in the squid giant synapse, sodium pentobarbital produced a dose-related, reversible decrease of the calcium spike of the presynaptic terminal.This work was performed while Dr Morgan was a Grass Foundation Fellow at the Marine Biological Laboratory, Woods Hole, Massachusetts, USA.The authors thank Dr J. Z. Yeh for the loan of the 4-aminopyridine and Dr G. H. Acheson for helpful suggestions.     

CDK5 downregulation enhances synaptic plasticity

CDK5 is a serine/threonine kinase that is involved in the normal function of the adult brain and plays a role in neurotransmission and synaptic plasticity. However, its over-regulation has been associated with Tau hyperphosphorylation and cognitive deficits. Our previous studies have demonstrated that CDK5 targeting using shRNA-miR provides neuroprotection and prevents cognitive deficits. Dendritic spine morphogenesis and forms of long-term synaptic plasticity—such as long-term potentiation (LTP)—have been proposed as essential processes of neuroplasticity. However, whether CDK5 participates in these processes remains controversial and depends on the experimental model. Using wild-type mice that received injections of CDK5 shRNA-miR in CA1 showed an increased LTP and recovered the PPF in deficient LTP of APPswe/PS1Δ9 transgenic mice. On mature hippocampal neurons CDK5, shRNA-miR for 12 days induced increased dendritic protrusion morphogenesis, which was dependent on Rac activity. In addition, silencing of CDK5 increased BDNF expression, temporarily increased phosphorylation of CaMKII, ERK, and CREB; and facilitated calcium signaling in neurites. Together, our data suggest that CDK5 downregulation induces synaptic plasticity in mature neurons involving Ca²⁺ signaling and BDNF/CREB activation.     

Developmental nicotine exposure induced alterations in behavior and glutamate receptor function in hippocampus

In the developing brain, nicotinic acetylcholine receptors (nAChRs) are involved in cell survival, targeting, formation of neural and sensory circuits, and development and maturation of other neurotransmitter systems. This regulatory role is disrupted when the developing brain is exposed to nicotine, which occurs with tobacco use during pregnancy. Prenatal nicotine exposure has been shown to be a strong risk factor for memory deficits and other behavioral aberrations in the offspring. The molecular mechanisms underlying these neurobehavioral outcomes are not clearly elucidated. We used a rodent model to assess behavioral, neurophysiological, and neurochemical consequences of prenatal nicotine exposure in rat offspring with specific emphasis on the hippocampal glutamatergic system. Pregnant dams were infused with nicotine (6 mg/kg/day) subcutaneously from the third day of pregnancy until birth. Results indicate that prenatal nicotine exposure leads to increased anxiety and depressive-like effects and impaired spatial memory. Synaptic plasticity in the form of long-term potentiation (LTP), basal synaptic transmission, and AMPA receptor-mediated synaptic currents were reduced. The deficit in synaptic plasticity was paralleled by declines in protein levels of vesicular glutamate transporter 1 (VGLUT1), synaptophysin, AMPA receptor subunit GluR1, phospho(Ser845) GluR1, and postsynaptic density 95 (PSD-95). These results suggest that prenatal nicotine exposure by maternal smoking could result in alterations in the glutamatergic system in the hippocampus contributing to the abnormal neurobehavioral outcomes.     

Signaling in the Chemosensory Systems

Taste bud cells communicate with sensory afferent fibers and may also exchange information with adjacent cells. Indeed, communication between taste cells via conventional and/or novel synaptic interactions may occur prior to signal output to primary afferent fibers. This review discusses synaptic processing in taste buds and summarizes results showing that it is now possible to measure real-time release of synaptic transmitters during taste stimulation using cellular biosensors. There is strong evidence that serotonin and ATP play a role in cell-to-cell signaling and sensory output in the gustatory end organs.