Glutamate-induced long-term potentiation of the frequency of miniature synaptic currents in cultured hippocampal neurons

Malgaroli, Antonio; Tsien, Richard W.

doi:10.1038/357134a0

Cited by 392 publications

(215 citation statements)

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“…Our results agree with this suggestion. The results of another recent study are in accordance with our findings: in cultured CA3-CA1 neurons a sustained potentiation of the frequency of miniature synaptic currents following the application of the excitatory neurotransmitter glutamate was demonstrated in the absence of Ca entry into the presynaptic terminal (Malgaroli and Tsien, 1992). In addition, at mossy fiber-CA3 synapses, although being different from the investigated CA3-CA1 synapses in that the induction of LTP is NMDA receptor independent (for review, see Johnston et al, 1992), the maintenance of LTP has been reported to be independent of presynaptic residual Ca (Regehr and Tank, 1991b).…”

Section: Ca Indicator Loading Techniquesupporting

confidence: 81%

“…Since we did not measure quanta1 parameters, we cannot judge the relative contribution of presynaptic and postsynaptic enhancement during LTP in our experiments. However, accumulated evidence has shown that a substantial component of LTP can be accounted for by presynaptic enhancement in CA3-CA1 synapses (Bekkers and Stevens, 1990;Malinow and Tsien, 1990;Kullmann and Nicoll, 1992;Liao et al, 1992;Malgaroli and Tsien, 1992). Given such a presynaptic contribution to the maintenance of LTP, our results suggest that some intermediate step between the presynaptic Ca transient and transmitter release is the likely site of persistent change.…”

Section: Ca Indicator Loading Techniquementioning

confidence: 67%

“…LTP is generally thought to be involved in learning and memory. Multiple lines of evidence including results of quanta1 analysis (Bekkers and Stevens, 1990;Malinow and Tsien, 1990;Kullmann and Nicoll, 1992;Liao et al, 1992;Malgaroli and Tsien, 1992) and the demonstration of postsynaptically released presumed retrograde messengers (Kom and Faber, 199 1;Fazeli, 1992) strongly suggest that the presynaptic site participates in the maintenance of LTP in area CA 1 of the hippocampus. Three mechanisms may underlie a presynaptic maintenance of LTP:…”

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confidence: 99%

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Presynaptic calcium is increased during normal synaptic transmission and paired-pulse facilitation, but not in long-term potentiation in area CA1 of hippocampus

1994

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We examined the relationship between presynaptic calcium levels and postsynaptic potentials during normal synaptic transmission, paired- pulse facilitation (PPF), and long-term potentiation (LTP) in CA3-CA1 synapses of hippocampus. By selectively loading the presynaptic terminals with the calcium indicator fura-2, we simultaneously recorded a presynaptic calcium (Ca) transient and the corresponding field EPSP evoked by a single stimulus given to the Schaffer collateral- commissural pathway in guinea pig hippocampal slices. A volume average presynaptic Ca influx was obtained by taking the first time derivative of the Ca transient. Our data indicate that the synaptic transmission represented by the initial slope of the field EPSP is approximately proportional to the fourth power of the presynaptic Ca influx, the volume average Ca current. Our results in combination with similar findings at the squid giant synapse (Augustine et al., 1985b; Augustine and Charlton, 1986) suggest that the relationship between Ca influx and transmitter release is well conserved from the molluscan to the mammalian nervous system. A transient increase of the residual Ca level ([Ca]res) is generally thought to be the mechanism underlying PPF (Katz and Miledi, 1968; Charlton et al., 1982); however, the relationship between PPF and the presynaptic [Ca]res had not been examined before. Our results demonstrate that PPF is approximately linearly related to the [Ca]res. This finding further supports the residual Ca hypothesis for PPF. Accumulated evidence from other groups suggests that the presynaptic site contributes to the maintenance of LTP in CA3-CA1 synapses (Bekkers and Stevens, 1990; Malinow and Tsien, 1990); however, our data show that neither an increase of the Ca transient nor a sustained increase of the [Ca]res occurs in the presynaptic terminals during maintenance of LTP. This suggests that the presynaptic mechanism underlying LTP must be downstream to Ca influx.

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Section: Ca Indicator Loading Techniquesupporting

confidence: 81%

Section: Ca Indicator Loading Techniquementioning

confidence: 67%

mentioning

confidence: 99%

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Presynaptic calcium is increased during normal synaptic transmission and paired-pulse facilitation, but not in long-term potentiation in area CA1 of hippocampus

1994

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“…The preparation of the hippocampal cell cultures from 3-to 5-day-old neonatal Sprague-Dawley rats has been described in detail elsewhere (9,11,12). The protocol consists in an enzymatic treatment (3.4 mg͞ml trypsin type XI ϩ 0.9 mg͞ml DNase type IV) followed by a gentle mechanical dissociation in Ca 2ϩ -free Hanks' solution supplemented with 12 mM MgSO 4 , 0.4 mg͞ml DNase, and 3 mg͞ml BSA.…”

Section: Methodsmentioning

confidence: 99%

Muscarinic stimulation of synaptic activity by protein kinase C is inhibited by adenosine in cultured hippocampal neurons

Bouron

Réuter²

1997

Proc. Natl. Acad. Sci. U.S.A.

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We have studied the effect of the cholinergic agonist carbachol on the spontaneous release of glutamate in cultured rat hippocampal cells. Spontaneous excitatory postsynaptic currents (sEPSCs) through glutamatergic ␣-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type channels were recorded by means of the patch-clamp technique. Carbachol increased the frequency of sEPSCs in a concentration-dependent manner. The kinetic properties of the sEPSCs and the amplitude distribution histograms were not affected by carbachol, arguing for a presynaptic site of action. This was confirmed by measuring the turnover of the synaptic vesicular pool by means of the f luorescent dye FM 1-43. The carbachol-induced increase in sEPSC frequency was not mimicked by nicotine, but could be blocked by atropine or by pirenzepine, a muscarinic cholinergic receptor subtype M1 antagonist. Intracellular Ca 2؉ signals recorded with the f luorescent probe Fluo-3 indicated that carbachol transiently increased intracellular Ca 2؉ concentration. Since, however, carbachol still enhanced the sEPSC frequency in bis(2-aminophenoxy)ethane-N,N,N ,N -tetra-acetate-loaded cells, this effect could not be attributed to the rise in intracellular Ca 2؉ concentration. On the other hand, the protein kinase inhibitor staurosporine as well as a down-regulation of protein kinase C by prolonged treatment of the cells with 4␤-phorbol 12-myristate 13-acetate inhibited the carbachol effect. This argues for an involvement of protein kinase C in presynaptic regulation of spontaneous glutamate release. Adenosine, which inhibits synaptic transmission, suppressed the carbachol-induced stimulation of sEPSCs by a G proteindependent mechanism activated by presynaptic A1-receptors.The occurrence of memory deficits in neurodegenerative disorders, such as Alzheimer disease, or the effects of cholinergic drugs on learning and memory have revealed an important role of the central cholinergic system in human cognitive functions (1-4). The hippocampus, which is known to be involved in memory formation (1, 4), receives cholinergic neurons from the basal forebrain. The cholinergic septohippocampal pathway modulates the activity of the hippocampus. For instance it is involved in the control of the theta rhythmic activity (5).Hippocampal cells express cholinergic nicotinic and muscarinic receptors with a predominance of the latter (5). Five muscarinic cholinergic receptor subtypes (M1-M5) have been identified (6). M1, M3, and M5 are coupled to phospholipase C activation resulting in the production of the second messengers inositol trisphosphate (IP 3 ) and diacylglycerol, whereas M2 and M4 inhibit adenylate cyclase activity (6). Immunoprecipitation studies, measurements of mRNA levels and of ligand binding sites indicate that the M1-subtype is the most abundant muscarinic receptor in the hippocampus (7).However, the role of M1-receptor activation and the subsequent signaling pathway in neurotransmitter release are only poorly understood. We have studied the effect of ...

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“…The procedures for culturing the mouse and rat glial cells are similar 59,60 , but we have modified them slightly to accommodate for the more rapid in vitro growth of rat vs. mouse glial cells. The procedures for the rat cell culture were modified from [61][62][63] . The use of a glial feeder layer for low-density neuronal cultures makes it necessary to perform rapid genotyping, in order to match the genotype of the feeder layer to that of the neurons within the period between the pups' births and the neonatal deaths or the end of culture window (1-2 postnatal days).…”

Section: Neuronal Culturesmentioning

confidence: 99%

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons

Koh

Iwabuchi

Huang

et al. 2015

JoVE

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High-resolution analysis of the morphology and function of mammalian neurons often requires the genotyping of individual animals followed by the analysis of primary cultures of neurons. We describe a set of procedures for: labeling newborn mice to be genotyped, rapid genotyping, and establishing low-density cultures of brain neurons from these mice. Individual mice are labeled by tattooing, which allows for long-term identification lasting into adulthood. Genotyping by the described protocol is fast and efficient, and allows for automated extraction of nucleic acid with good reliability. This is useful under circumstances where sufficient time for conventional genotyping is not available, e.g., in mice that suffer from neonatal lethality. Primary neuronal cultures are generated at low density, which enables imaging experiments at high spatial resolution. This culture method requires the preparation of glial feeder layers prior to neuronal plating. The protocol is applied in its entirety to a mouse model of the movement disorder DYT1 dystonia (ΔE-torsinA knock-in mice), and neuronal cultures are prepared from the hippocampus, cerebral cortex and striatum of these mice. This protocol can be applied to mice with other genetic mutations, as well as to animals of other species. Furthermore, individual components of the protocol can be used for isolated sub-projects. Thus this protocol will have wide applications, not only in neuroscience but also in other fields of biological and medical sciences. Video LinkThe video component of this article can be found at

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Glutamate-induced long-term potentiation of the frequency of miniature synaptic currents in cultured hippocampal neurons

Cited by 392 publications

References 50 publications

Presynaptic calcium is increased during normal synaptic transmission and paired-pulse facilitation, but not in long-term potentiation in area CA1 of hippocampus

Presynaptic calcium is increased during normal synaptic transmission and paired-pulse facilitation, but not in long-term potentiation in area CA1 of hippocampus

Muscarinic stimulation of synaptic activity by protein kinase C is inhibited by adenosine in cultured hippocampal neurons

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons

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