Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate

Burger, Peter; Mehl, E.; Cameron, Patricia L.; Maycox, Peter R.; Baumert, Marion; Lottspeich, Friedrich; Camilli, Pietro De; Jahn, Reinhard

doi:10.1016/0896-6273(89)90240-7

Cited by 286 publications

(235 citation statements)

References 26 publications

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“…Thus in contrast to the numerous reports of glutamate uptake into synaptic vesicle preparations Ueda, 1983,1985;Fischer-Bovenkerk et al, 1988;Maycox et al, 1988;Shioi et al, 1989;Kish et al, 1989a;Carlson et al, 1989;Shioi and Ueda, 1990;Lobur et al, 1990) no evidence for vesicular aspartate uptake has been presented. Furthermore, synaptic vesicles isolated in the presence of N-ethylmaleimide contain no detectable aspartate (Burger et al, 1989). This is in interesting contrast to the pair of inhibitory amino acids,…”

Section: The Vesicular Transporter and Discrimination Between Glutamacontrasting

confidence: 39%

“…Conditions suitable for the isolation of vesicles containing the latter transmitters (hypotonic lysis of synaptosomes followed by a 2-h centrifugation in the absence of ATP) produce vesicles containing no significant GABA or glutamate (DeBelleroche and Bradford, 1977). The availability of rapid immuno-purification techniques for synaptic vesicles (Naito and Ueda, 1983;Burger et al, 1989) and the realization that the leakage of amino acids from vesicles can be prevented by including ATP in the preparation medium to prevent the collapse of the vesicular proton gradient and high concentrations of Nethylmaleimide to block the vesicular transporters, has allowed the isolation of vesicles containing an estimated 50-1OOmM glutamate (Burger et al, 1989). It must of course be remembered that these preparations of synaptic vesicles are not homogeneous with respect to neurotransmitter, but reflect the heterogeneity of the brain area from which they were prepared.…”

Section: Glutamate Accumulation Into Small Synaptic Vesiclesmentioning

confidence: 99%

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The glutamatergic nerve terminal

Nicholls

1993

European Journal of Biochemistry

208

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Glutamate as a neurotransmitterThe human brain has about lolo neurones, each of which, on average, makes about 1000 synaptic contacts with other neurones. Of these l O I 3 synapses perhaps up to 90% utilize amino acids as their neurotransmitter. Glutamate is the major excitatory amino acid (acting predominantly on depolarizing post-synaptic receptors) while 4-aminobutyrate (GABA) and glycine are the major inhibitory transmitters (acting on hyper-polarizing receptors). As well as playing the dominant role in fast information transfer, glutamate is of particular interest in view of its involvement in current models of memory and learning (Bliss and Dolphin, 1982;Cotman et al., 1988;Collingridge and Singer, 1990) and because a pathological release of glutamate in brain ischaemia is neurotoxic and a major contributor to the damage caused under these conditions (Rothman and Olney, 1986;Choi, 1988 ;Meldrum and Garthwaite, 1990).A synapse has a pre-synaptic element responsible for the storage and release of transmitter and a post-synaptic element containing one or more classes of receptor for the transmitter. The potent combination of electrophysiology and molecular cloning has allowed a dramatic advance in our understanding of post-synaptic events. Three classes of post-synaptic ionotropic (possessing an integral ion channel) glutamate receptors have been identified and cloned: the 2-amino-3-hydroxy-5-methyl-4-isoxazole propionatekainate (AMPA/ KA) receptor Nakanishi et al., 1990;Sakmann, 1992), the high-affinity KA receptor (Egebjerg et al., 1991) and the N-methyl D-aSpartate (NMDA) receptor (Moriyoshi et al., 1991 ; Kumar et al., 1991 ;Barnard, 1992). Each can exist in many isoforms by combining subunits formed from distinct genes, by alternative splicing or by post-transcriptional modification (Monyer et al., 1991 ;Burnashev et al., 1992).The AMPNKA receptor is responsible for fast information transfer while the NMDA receptor is only operative when the post-synaptic membrane is independently depolarized by another receptor. This 'associative' behaviour of the latter is believed to underlie aspects of the learning process.

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Section: The Vesicular Transporter and Discrimination Between Glutamacontrasting

confidence: 39%

Section: Glutamate Accumulation Into Small Synaptic Vesiclesmentioning

confidence: 99%

The glutamatergic nerve terminal

Nicholls

1993

European Journal of Biochemistry

208

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“…To discriminate between these two possibilities, SSV immunoisolation experiments were carried out. This strategy, successfully used in the past to isolate glutamate-containing vesicles from brain homogenates, was chosen because it does not perturb the integrity of the SSV by avoiding the presence of detergents (52). In this condition, the N-terminal directed Syt I antibody presents its limits due to the inaccessibility of the lumen of the SSV.…”

Section: Resultsmentioning

confidence: 99%

Calcium-dependent Oligomerization of Synaptotagmins I and II

Osborne

Herreros

Bastiaens

et al. 1999

Journal of Biological Chemistry

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Synaptotagmins constitute a large family of membrane proteins characterized by their distinct distributions and different biochemical features. Genetic evidence suggests that members of this protein family are likely to function as calcium sensors in calcium-regulated events in neurons, although the precise molecular mechanism remains ill defined. Here we demonstrate that different synaptotagmin isoforms (Syt I, II, and IV) are present in the same synaptic vesicle population from rat brain cortex. In addition, Syt I and II co-localize on the same small synaptic vesicle (SSV), and they heterodimerize in the presence of calcium with a concentration dependence resembling that of the starting phase of SSV exocytosis (EC 50 ‫؍‬ 6 ؎ 4 M). The association between Syt I and Syt II was demonstrated by immunoprecipitation of the native proteins and the recombinant cytoplasmic domains and by using fluorescence resonance energy transfer (FRET). Although a subpopulation of SSV containing Syt I and IV can be isolated, these two isoforms do not show a calcium-dependent interaction. These results suggest that the self-association of synaptotagmins with different calcium binding features may create a variety of calcium sensors characterized by distinct calcium sensitivities. This combinatorial hypothesis predicts that the probability of a single SSV exocytic event is determined, in addition to the gating properties of the presynaptic calcium channels, by the repertoire and relative abundance of distinct synaptotagmin isoforms present on the SSV surface.Neuronal communication depends upon the transduction of an electric nerve impulse for the release of neurotransmitters from their storage compartment, the small synaptic vesicles (SSV).1 This tightly regulated process is triggered by the rapid increase of the intracellular calcium due to the opening of voltage-gated calcium channels at the active zones of the synaptic plasma membrane, which causes the activation of the SSV fusion machinery (1, 2). Of the proteins involved in the physiological cycle of the SSV, the members of the synaptotagmin family represent the best candidates for the role of calcium sensors in neurotransmitter release and, more generally, in regulated exocytosis (3-5). This protein family comprises more than a dozen members having the same overall structure and a variable degree of homology (4, 5). Synaptotagmins are integral membrane proteins with broad distributions in neuronal and non-neuronal tissues that vary between isoforms (4, 5). In the central nervous system and neuroendocrine cells, synaptotagmins are localized on SSV and secretory granules (6). These proteins are characterized by a single membrane-spanning domain and by a large cytoplasmic portion containing two internal repeats that have homology with the C2 domain of protein kinase C (7,8). This domain is known to regulate the calciumdependent translocation of protein kinase C to membranes and is present in many proteins with different functions (5, 8). In synaptotagmins, the two C2 domains are re...

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“…The concentration of glutamate in the vesicle is estimated to be on the order of 100 mM (Burger et al, 1989). Vesicle diameter has been measured by EM to be 29-36 nm in goldfish bipolar cell terminals Lagnado et al, 1996) but 45-50 nm in diameter in salamander photoreceptor terminals (Lasansky, 1973;Thoreson et al, 2004).…”

Section: Synaptic Vesiclesmentioning

confidence: 99%

Synaptic transmission at retinal ribbon synapses

Heidelberger

Thoreson

Witkovsky

2005

Progress in Retinal and Eye Research

209

231

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The molecular organization of ribbon synapses in photoreceptors and ON bipolar cells is reviewed in relation to the process of neurotransmitter release. The interactions between ribbon synapseassociated proteins, synaptic vesicle fusion machinery and the voltage-gated calcium channels that gate transmitter release at ribbon synapses are discussed in relation to the process of synaptic vesicle exocytosis. We describe structural and mechanistic specializations that permit the ON bipolar cell to release transmitter at a much higher rate than the photoreceptor does, under in vivo conditions. We also consider the modulation of exocytosis at photoreceptor synapses, with an emphasis on the regulation of calcium channels.

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Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate

Cited by 286 publications

References 26 publications

The glutamatergic nerve terminal

The glutamatergic nerve terminal

Calcium-dependent Oligomerization of Synaptotagmins I and II

Synaptic transmission at retinal ribbon synapses

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