[Show abstract][Hide abstract] ABSTRACT: Syntaxin, a protein required for the docking of synaptic vesicles, may be involved in the manifestation of synaptic plasticity. The possible involvement of syntaxin in epileptogenesis was investigated by assessing the expression levels of syntaxin1B/GR33 mRNA by in situ hybridization at different stages of hippocampal kindling epileptogenesis and after the induction of generalized seizures. Densitometric analysis of the autoradiograms revealed that the expression was not changed in pyramidal and granular neurons of the hippocampal formation 24 h after the first kindling stimulation. However, the mRNA levels in CA1, CA3, and fascia dentata neurons were bilaterally enhanced after six afterdischarges and remained at this elevated level during the whole period along which afterdischarges were elicited. An immunoassay was unable to reveal a clear significant increase of syntaxin1B/GR33 protein levels in hippocampus homogenates of fully kindled animals. The use of syntaxin1B-specific antibodies is necessary to draw definite conclusions on the changes at the protein level. At long term, 4 weeks after the last kindling-elicited generalized seizure, no significant alterations in transcript levels could be detected. The results suggest that the induction of kindling epileptogenesis is associated with an enhanced expression of syntaxin1B/GR33, but this enhanced expression is not necessary for persistence of kindling-induced synaptic plasticity.
Journal of Neurochemistry 12/1995; 65(5):1974-80. DOI:10.1046/j.1471-4159.1995.65051974.x · 4.24 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Polyclonal antibodies raised against rat syntaxin-1B and an affinity-purified fraction have been used to study the functional role of this protein in transmitter release from Aplysia neurons. In a ganglionic protein extract, this fraction recognized a 37,000 molecular weight protein which therefore might be the Aplysia homologue of rat brain syntaxin-1B. Immunoglobulins were injected in the presynaptic cell of an identified cholinergic synapse of the buccal ganglion of Aplysia californica. This treatment decreased the postsynaptic response due to a reduction of the number of quanta released in relation to a decline of presynaptic Ca2+ current. When antibodies were applied extracellularly, transmitter release also decreased. In contrast to intracellular injection, this reduction was not accompanied by a decrease of the Ca2+ current but by an increase of presynaptic outward current. When injected in the presynaptic neuron, syntaxin complementary RNA also depressed Ca2+ current and transmission. This work provides evidence that Aplysia neurons express a syntaxin-like protein which is involved in the control of the presynaptic Ca2+ influx triggering acetylcholine release from terminals. This protein appears to have an extracellular segment which might interact with outward current.
[Show abstract][Hide abstract] ABSTRACT: Glutamate receptors mediate excitatory neurotransmission in the brain and are important in the formation of memory and in some neurodegenerative disorders. A complementary DNA clone that encoded a 33-kilodalton protein (GR33) was obtained by screening a library with an antibody generated against glutamate binding proteins. The sequence of GR33 is identical to that of the recently reported presynaptic protein syntaxin. When GR33 was expressed in Xenopus oocytes, it formed glutamate-activated ion channels that are pharmacologically similar to those of N-methyl-D-aspartate receptors but with different electrophysiological properties. Mutation of the leucine 278 residue in the single putative transmembrane segment of GR33 affects the properties of the channel. Thus, in vivo GR33 may be a presynaptic glutamate receptor.
[Show abstract][Hide abstract] ABSTRACT: Repetitive activation of excitatory synapses in the hippocampus produces a persistent enhancement of synaptic efficiency known as long-term potentiation (LTP). In anesthetized and in freely moving rats, the induction of LTP in the perforant path led to a transient increase in the amount of messenger RNA (mRNA) coding for a presynaptic glutamate receptor (GR33) in dentate granule cells. The amount of GR33 mRNA was increased for at least 5 hours after the induction of LTP but was indistinguishable from control values 1 day after induction. The N-methyl-D-aspartate receptor antagonist 2-aminophosphonovalerate prevented the induction of both LTP and the increase in GR33 mRNA. The amount of GR33 protein was increased in the mossy fiber terminal zone of dentate granule cells 5 hours after the induction of LTP. These results suggest that the induction of LTP in synapses at one stage in a neural network may lead to modification in synaptic function at the next stage in the network.