Molecular mechanisms of dendritic spine development and maintenance

The large majority of excitatory synapses are located on dendritic spines which are discrete membrane protrusions present on neuronal dendrites. Interestingly the highly heterogeneous morphology of dendritic spines is thought to be the morphological basis for synaptic plasticity associated to learning and memory formation. Indeed dendritic spines structure is regulated by molecular mechanisms that are fine tuned and adjusted according to level and direction of synaptic activity, development, specific brain region, and different experimental behavioral conditions. This supports the idea that reciprocal changes between the structure and function of spines impact both local and global integration of signals within dendrites. An increasing number of proteins have been found to be morphogens for dendritic spines and provided new insights into the molecular mechanisms regulating spine formation and morphology. Thus determining the mechanisms that regulate spine formation and morphology is essential for understanding the cellular changes that underlie learning and memory in normal and pathological conditions.     

Excitatory amino acid neurotoxicity in the hippocampal slice preparation

The effect of sustained activation of excitatory amino acid receptors on neuronal survival was studied using slices of adult rat hippocampus and light and electron microscopy. Kainate, N-methyl-D-aspartate, quisqualate, and ibotenate all produce signs of severe neurotoxicity within 90 min. Neuronal damage occurs in the form of perikaryal and dendritic swelling, cytoplasmic and nucleoplasmic disintegration, and plasma and nuclear membrane ruffling and collapse. The toxicity is restricted to intrinsic neuronal somata, dendrites and spines, while afferent axons, boutons and glia are spared. Although damage is generally distributed throughout all areas of hippocampus, kainate has little effect on pyramidal neurons in the CA2 region. Quantitative analysis of neuronal survival indicates that agonists induce dose-dependent damage over concentration ranges known to be excitatory. Based on selective antagonism by DL-aminophosphonoheptanoate and the patterns of damage produced by each, N-methyl-D-aspartate, kainate, and quisqualate trigger neurotoxicity by acting on distinct receptor classes. It is concluded that, in hippocampal slices, excitatory amino acids induce neurotoxicity in a similar manner to their actions in vivo. The results support the hypothesis that hippocampal neurotoxicity is initiated by excessive excitation, and provide another example of the capacity of adult hippocampal neurons for rapid structural modification.     

Excitatory amino acid receptors and ischemic brain damage in the rat

The excitatory amino acid glutamate has been suggested to be an important mediator of the selective CA1 hippocampal damage which follows transient cerebral ischemia. In order to evaluate the possible involvement of altered glutamate receptor regulation in the expression of the delayed neuronal necrosis following ischemia, we have determined the density of glutamate receptor subtypes in the rat hippocampus following transient ischemia. We report a transient reversible decrease in [3H]AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) binding sites (presumably representing quisqualate receptors) followed by a long term loss of binding at 2 days postischemia which precedes neuronal loss. In contrast, no change was noted in the N-methyl-D-aspartate or kainic acid binding sites over this time period.     

Excitatory amino acid modulation of lordosis in the rat.

Microinfusion of the excitatory amino acid agonist N-methyl-D-aspartate (NMDA) into the mediobasal hypothalamus (MBH) significantly reduced lordosis in estrogen plus progesterone-treated female rats at 10 min post-infusion with recovery to pretest values by 30 min (P less than .05; Wilcoxon). Microinfusion of the specific NMDA antagonist D,L2-amino-5-phosphonopentoic acid (AP-5) into the same sites was without effect on lordosis responding of fully receptive females. There was also a significant increase in the number of females vocalizing to mounts by males after infusion of NMDA but not after infusion of AP-5 into the MBH. When NMDA was infused into the preoptic area (POA) there was no effect on lordosis responding of full receptive females, but AP-5 infusion resulted in a significant inhibition of lordosis at 10 min post-infusion. There was no difference between groups in percentage of females vocalizing after drug infusion into the POA. These results suggest that increased excitatory amino acid activity in the MBH and decreased excitatory amino acid activity in the POA inhibits lordosis behavior.     

Role of APP for dendritic spine formation and stability

The amyloid precursor protein (APP) is transported in high amounts to the presynaptic endings where its function is still unknown. Several studies indicate that lack of APP or its overexpression affects the number of dendritic spines, the postsynaptic compartment of excitatory synapses. Since synapse loss has been identified as one of the most important structural correlates of cognitive decline in Alzheimer's diseases (AD), the physiological function of APP at synapses, specifically at dendritic spines, has come into focus in AD research. This review intends to give an overview of the very controversial results on APP expression on dendritic spine number in the mouse brain.     

Excitatory amino acid receptors mediate slow synaptic transmission in turtle cerebellum

In the isolated turtle cerebellum intracellular recordings from Purkinje cell dendrites and somata reveal novel slow excitatory synaptic potentials evoked by activation of climbing fiber (CF) or parallel fiber (PF) inputs. Classical fast excitatory synaptic responses to CF and PF stimulation are followed by large, slow excitatory postsynaptic potentials (sEPSPs) which are associated with an increase in conductance and are enhanced by hyperpolarization. Both sEPSPs are blocked by the excitatory amino acid (EAA) antagonist kynurenate, but not by DL-2-amino-5-phosphonovalerate (AP-5). The EAA receptor antagonist L-amino-4-phosphonobutyric acid (L-AP-4) reversibly blocked the PF-sEPSP without affecting the CF-sEPSP. Two novel slow synaptic potentials mediated by excitatory amino acid receptors can therefore be observed in turtle cerebellum which may play an important role in synaptic integration.     

Excitatory amino acid receptors in the prefrontal cortex of aging mice

The zones of the prefrontal cortex of Balb/c mice were tested for age-related changes of the ionotropic excitatory amino acid receptors density, together with zones of the dorsal cortex. Kainate, N-methyl-D-aspartate, and amino-3-hydroxy-5-methyloxazole-4-propionate sites were measured by slice receptor binding techniques in cortical zones from animals at the age of 6, 12, 18, and 24 months. An increase of the N-methyl-D-aspartate sites was detected in the medial prefrontal zone of mid-aged animals and was followed by a decrease at old age; a decrease of the N-methyl-D-aspartate and kainate sites was found for the medial dorsal (cingulate) cortex at old age. The age-related changes of receptor densities in the different cortical areas seem unrelated in origin. The sites decrease in the cingulate cortex could affect the transfer of the prefrontal cortex activity toward limbic structures.     

Excitatory amino acids in the chick retina: possible involvement of cyclic guanosine monophosphate

An investigation has been made of the ability of excitatory amino acids to alter the levels of chick retinal cyclic guanosine monophosphate (GMP). Kainic acid, N-methyl-D-aspartic acid and quisqualic acid each produced dose-related increases in cyclic GMP, and these could be attenuated by the addition of excitatory amino acid antagonists. It appears likely that in addition to its proposed role in visual transduction, this nucleotide may be involved in excitatory amino acid function in the chick retina.     

Drebrin在神经元树突棘形成和重塑中的作用

drebrin是一种与脑发育相关的调节蛋白,由Shirao等人于1986年从鸡胚中发现[1].随后的研究使人们对其结构、分布、功能及机制等方面有了全面认识.人们发现drebfin可与纤维型肌动蛋白(F-actin)结合和解离,具有肌动蛋白结合蛋白的特征,并能影响神经元树突棘内其它肌动蛋白结合蛋白的分布,改变树突棘的形态,从而影响突触的结构和功能.drebrin的这一功能,在神经突起的形成、突触的形成和重建过程中发挥重要的作用.鉴于目前国内对drebrin的研究甚少,本文对drebrin的相关研究结果,特别是drebrin在神经元树突棘形成和重塑中的作用进行了综述.     

Excitatory amino acid transporter EAAT-2 in tangle-bearing neurons in Alzheimer's disease

The excitatory amino acid transporter EAAT-2 is physiologically expressed in astrocytes. This study demonstrates that distinct subclasses of neurons exhibited EAAT-2 immunoreactivity in cases with Alzheimer's disease (AD). EAAT-2 was identified in the following types of neurons: Cortical pyramidal cells, fascia dentata granule cells, neurons of the basal nucleus of Meynert, the substantia nigra, the paraventricular nucleus of the hypothalamus, oral and central raphe nuclei, locus coeruleus, parabrachial nucleus, and neurons of the reticular formation of the brain stem. All EAAT-2-positive neurons displayed cytoskeletal abnormalities with abnormal tau-protein and often showed condensed and shrunken nuclei. None of the control cases without AD-related pathology showed EAAT-2-immunoreactive neurons. These results indicate that AD-related neurodegeneration is associated with the expression of the glutamate transporter EAAT-2 in altered neurons. Since an aberrant expression of EAAT-1 in neurons has recently been described, the finding of a neuronal expression of EAAT-2 strongly supports the hypothesis that abnormalities in glutamate transport play an important role in the pathogenesis of AD.     

Excitatory amino acid receptors in brains of rats with methylazoxymethanol-induced microencephaly.

We used methylazoxymethanol-acetate (MAM), a potent alkylating agent, to produce microencephaly in offspring by injecting it into pregnant rats on day 15 of gestation. Binding activities of central excitatory amino acid receptors were examined in Triton-treated membranes prepared from brains of adult offspring with MAM-induced microencephaly (MAM rats). MAM rats exhibited approximately 40-50% reductions of the wet weights of the cerebral cortex, hippocampus and striatum compared to those in controls. In the cortex and hippocampus of MAM-rats, total bindings of [3H]glutamate (Glu) (which is sensitive to N-methyl-D-aspartate (NMDA) receptor), and strychnine-insensitive [3H]glycine (Gly) and (+)-5-[3H]methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imi ne (MK-801; a noncompetitive antagonist of NMDA receptor), were reduced to approximately 40% of those in controls. Similarly, in both regions of MAM rats, total bindings of [3H]kainate and DL-alpha-amino-3-[3H]hydroxy-5-methylisoxazole-4-propionic acid (an agonist of quisqualate receptors), were reduced to approximately 35-50% of those in controls. However, total bindings of these radioligands in the striatum of MAM rats were more than 65% of those in controls, despite the significant loss of striatum mass. However, specific bindings of radioligands in the striatum of MAM rats were elevated by more than 60% of those in controls, and Scatchard analysis revealed that elevations of [3H]Glu, [3H]Gly and [3H]MK-801 bindings were due to a significant increase in the densities of binding sites, with their affinities remaining unaltered. Spatial recognition ability examined by an 8-armed radial maze task was markedly impaired compared to those in controls. These results suggest that the proliferation of neurons bearing excitatory amino acid receptors (EAA) in the striatum is less affected by MAM treatment on day 15 of gestation than that in the cortex and hippocampus in spite of drastic weight loss in these brain regions. The significant reduction of EAA receptors in the cortex and hippocampus may be involved in the impairment of spatial memory observed in MAM-treated rats.     

Excitatory amino acid transmitters in neuronal circuits of the cat visual cortex

To test a possibility that glutamate (Glu) and aspartate (Asp) are transmitters in the visual cortex and to locate their operating sites in the cortical circuitry, we studied effects of microiontophoretic application of Glu/Asp antagonists on visual responses of cortical neurons in the cat. The antagonists tested were kynurenic acid (KYNA), cis-2,3-piperidine dicarboxylic acid, and gamma-D-glutamylglycine. Among these antagonists, KYNA was most effective in blocking visual responses of cortical neurons; it eliminated visual responses in 156 of the 188 cells tested. Usually the maximal suppressive effect appeared 20-30 s after starting KYNA application and recovery of cell's responsiveness 30-60 s after stopping the application. KYNA antagonized excitations induced by ionophoretic application of Glu and Asp but did not block those by acetylcholine, suggesting that KYNA is a selective antagonist of Glu/Asp, and its action is not due to general depressant effects. This suggestion was further supported by the observation that in corticogeniculate cells the latency and probability of invasion of antidromic spikes into the somatodendritic part following electrical stimulation of the lateral geniculate were not changed while visual responses were completely suppressed by KYNA. In terms of actions of the three agonists which give the basis for classifying excitatory amino acid receptors into at least three types, KYNA antagonized excitations by N-methyl-D-aspartic acid (NMDA) and kainate in almost all the cells tested but did not block those by quisqualate in about half of the cells. These results suggest that KYNA reacts more preferentially with NMDA and kainate receptors than with quisqualate receptors. Effectiveness of KYNA was related to types of receptive fields of cells and to their laminar locations. In 79 of the 104 simple cells tested, KYNA completely suppressed their visual responses, while such a complete block was seen in only 18 of the 68 complex and 3 of the 16 special complex cells. The great majority of the cells in layers IVab, IVc and the upper part of layer VI were completely suppressed by KYNA, whereas most of the cells in the other layers were incompletely suppressed or not suppressed at all.(ABSTRACT TRUNCATED AT 400 WORDS)     

Excitatory amino acid binding sites in the periaqueductal gray of the rat

We used receptor autoradiography to determine the distribution of excitatory amino acid (EAA) binding site subtypes in the periaqueductal gray (PAG) of the rat. N-Methyl-D-aspartate (NMDA), kainate, quisqualate-ionotropic, and quisqualate-metabotropic binding sites were all present in the PAG. Distribution was inhomogeneous with greatest density of all binding site subtypes in the dorsolateral subdivision and lowest density in the ventrolateral subdivision. Relative to regions of brain with high densities of EAA binding site subtypes, quisqualate-metabotropic binding sites had the highest relative density and NMDA binding sites the least. The presence of all subtypes of EAA binding sites in the PAG suggests that EAA action within the PAG is likely to be complex.     

Excitatory amino acids and cerebrovascular tone

Levels of excitatory amino acids in the brain extracellular fluid compartment rise during pathological conditions in the brain such as ischaemia, anoxia and epilepsy. One such amino acid, glutamate, is present in sensory nerve fibres innervating, for example, cerebral vessels. Enhanced levels of circulating glutamate and aspartate are found in migraine sufferers. The present study examined whether excitatory amino acids, in concentrations found in the brain extracellular fluid compartment during pathological conditions, exert a direct effect on cerebrovascular tone. As tested in flow-regulating pial arteries from rat, cat and man, no such constrictive or dilatory effect was obtained.     

Excitatory amino acid receptors of rod- and cone-driven horizontal cells in the rabbit retina

Intracellular recordings were obtained from horizontal cells in the superfused retina-eyecup preparation of the rabbit. Rod- and cone-dominated horizontal cells were studied using bath-applied excitatory amino acid analogues. Cone-dominated horizontal cell somas were depolarized by kainate (KA) or quisqualate (QQ) and their light responses were reduced or abolished. They were not affected by N-methyl-DL-aspartate (NMDLA) at concentrations up to 2 mM or by 2-amino-4-phosphonobutyrate (APB), a selective agonist for the ON bipolar cell. When synaptic transmission was blocked with cobalt, horizontal cell somas were hyperpolarized. Under these conditions, KA and QQ caused large depolarizations suggesting that these agents have a direct action on horizontal cell somas. Excitatory amino acid antagonists such as cis-2,3-piperidine dicarboxylic acid (PDA) and kynurenic acid (Kyn) hyperpolarized horizontal cell somas to the level of the light-driven membrane potential. These antagonists blocked both the light-driven responses and the depolarizing action of KA. The specific NMDA antagonist 2-amino-7-phosphonoheptanoate (AP-7) had no effect on the membrane potential or light-driven responses of horizontal cell somas. In contrast to a previous report, we found no evidence that low concentrations of NMDLA could hyperpolarize horizontal cells or act as a KA antagonist in the rabbit retina. Rod-dominated axon terminals were identified by waveform, threshold, and the presence of a large rod after-potential evoked by high light intensity. These cells were depolarized by KA and their light responses were attenuated. NMDLA and APB had no effect on these cells. The general antagonists, PDA and Kyn, hyperpolarized axon terminals and blocked their light-evoked responses. The specific NMDA antagonist, AP-7, had no effect on these cells. These results suggest that the synaptic receptors that mediate light input to both rod- and cone-dominated horizontal cells are kainate or quisqualate receptors. This implies that the rod and cone transmitters of the rabbit retina are similar, with the characteristics of an excitatory amino acid, such as glutamate.     

Excitatory amino acid transmitters and their receptors in neural circuits of the cerebral neocortex

In 1954, L-glutamate (Glu) and L-aspartate (Asp) were first suggested as being excitatory synaptic transmitters in the cerebral cortex. Since then, evidence has mounted steadily in favor of the view that Glu and Asp are major excitatory transmitters in the neocortex. Many of the experimental studies which reported how Glu/Asp came to satisfy the criteria for transmitters in the neocortex are reviewed here, according to the methods employed. Since the question of which particular synaptic sites in cortical neural circuits Glu/Asp operate as excitatory transmitters has not previously been reviewed, particular attention is given to efferent, afferent and intrinsic neural circuits of the visual and somatosensory cortices, where circuitry is relatively clearly delineated. Recent studies using chemical assays of released amino acids, high-affinity uptake mechanisms of Glu/Asp from nerve terminals, the direct micro-iontophoretic administration of Glu/Asp antagonists, and immunocytochemical techniques have demonstrated that almost all corticofugal efferent projections employ Glu/Asp as excitatory synaptic transmitters. Evidence indicating that thalamocortical afferent projections, including geniculocortical projections and some intrinsic connections are glutamatergic, is also reviewed. Thus, the results highlighted here indicate that the main framework of neocortical circuitry is operated by Glu/Asp. Pharmacological studies indicate that synaptic receptors for Glu/Asp can be classified into a few subtypes, including N-methyl-D-aspartate (NMDA) and quisqualate/kainate (non-NMDA) types. Some evidence indicating the sites of operation of NMDA and non-NMDA receptors in neocortical circuitry is reviewed, and the distinct, functional significance of these two types of Glu/Asp receptors in information processing in the neocortex is proposed.     

Excitatory amino acid receptors in the parallel fibre pathway in rat cerebellar slices

The grease-gap technique was used on young rat cerebellar slices to study the synaptic pharmacology of the parallel fibre pathway. Electrical stimulation of the parallel fibres produced a characteristic response in Purkinje cells: a sharp negative (N) potential, representing the population action potential and underlying parallel fibre EPSP, followed by a slow positive (P) wave, the population inhibitory postsynaptic potential (IPSP). In the presence of 1.2 mM Mg2+, D-2-amino-5-phosphonovalerate (APV, 30 microM) had no effect but both potentials could be inhibited by 6-cyano-2,3-dihydroxy-7-nitro-quinoxaline (CNQX, 10 microM). Removal of Mg2+ had no effect on the N-potential but enhanced the P-wave in an APV-sensitive fashion, particularly when CNQX was present. The results provide further evidence that glutamate is the parallel fibre transmitter and suggest that its acts only on non-NMDA (non-N-methyl-D-aspartate) receptors at synapses with Purkinje cells but on both NMDA and non-NMDA receptors at synapses with inhibitory interneurones. At the latter synapses, the NMDA system is likely to be brought into operation in an activity-dependent manner.     

Excitatory amino acids and Alzheimer's disease

Excitatory amino acids (EAA) such as glutamate and aspartate are major transmitters of the cerebral cortex and hippocampus, and EAA mechanisms appear to play a role in learning and memory. Anatomical and biochemical evidence suggests that there is both pre- and postsynaptic disruption of EAA pathways in Alzheimer's disease. Dysfunction of EAA pathways could play a role in the clinical manifestations of Alzheimer's disease, such as memory loss and signs of cortical disconnection. Furthermore, EAA might be involved in the pathogenesis of Alzheimer's disease, by virtue of their neurotoxic (excitotoxic) properties. Circumstantial evidence raises the possibility that the EAA system may partially determine the distribution of pathology in Alzheimer's disease and may be important in producing the neurofibrillary tangles, RNA reductions and dendritic changes which characterize this devastating disorder. In this article, we will review the evidence suggesting a role for EAA in the clinical manifestations and pathogenesis of Alzheimer's disease.