A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system

Edwards, Francis A.; Konnerth, Arthur; Sakmann, Bert; Takahashi, Tomoyuki

doi:10.1007/bf00580998

Cited by 1,243 publications

(742 citation statements)

References 23 publications

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“…Presynaptic action potential and voltage-gated Na + and K + channels in hippocampal mossy fiber boutons After the development of methods for patch-clamp recording in brain slices [26,38], techniques were gradually refined to a point when it became possible to obtain recordings from presynaptic elements, including mossy fiber boutons [12,28,34]. The key steps of improvement were as follows: (1) the enhanced visibility provided by high-numerical aperture objectives and infrared illumination [75]; (2) the development of vibratomes with minimal blade vibration in the Fig.…”

Section: Abstract Presynaptic Recording Mossy Fiber Boutons Mossymentioning

confidence: 99%

Timing and efficacy of transmitter release at mossy fiber synapses in the hippocampal network

Bischofberger

Engel

Frotscher

et al. 2006

Pflugers Arch - Eur J Physiol

115

110

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It is widely accepted that the hippocampus plays a major role in learning and memory. The mossy fiber synapse between granule cells in the dentate gyrus and pyramidal neurons in the CA3 region is a key component of the hippocampal trisynaptic circuit. Recent work, partially based on direct presynaptic patch-clamp recordings from hippocampal mossy fiber boutons, sheds light on the mechanisms of synaptic transmission and plasticity at mossy fiber synapses. A high Na + channel density in mossy fiber boutons leads to a large amplitude of the presynaptic action potential. Together with the fast gating of presynaptic Ca 2+ channels, this generates a large and brief presynaptic Ca 2+ influx, which can trigger transmitter release with high efficiency and temporal precision. The large number of release sites, the large size of the releasable pool of vesicles, and the huge extent of presynaptic plasticity confer unique strength to this synapse, suggesting a large impact onto the CA3 pyramidal cell network under specific behavioral conditions. The characteristic properties of the hippocampal mossy fiber synapse may be important for pattern separation and information storage in the dentate gyrus-CA3 cell network.Keywords Presynaptic recording . Mossy fiber boutons . Mossy fiber synapses . Hippocampus . Autoassociative networks . Episodic memory . Synaptic efficacyThe mossy fiber synapse: a key connection in the hippocampal networkThe hippocampal formation consists of three types of principal neurons: granule cells in the dentate gyrus, CA3 pyramidal cells, and CA1 pyramidal neurons. These cells are interconnected by glutamatergic synapses, forming the classical trisynaptic circuit (Fig. 1a,b). Dentate gyrus granule cells receive excitatory glutamatergic input from layer 2 pyramidal cells of the entorhinal cortex and project to CA3 pyramidal cells. CA3 pyramidal cells project to CA1 cells, which in turn project to the subiculum and back to the entorhinal cortex [4]. In addition to this trisynaptic circuit, there is also a direct input from the entorhinal cortex to both CA3 and CA1 pyramidal cells. Furthermore, CA3 pyramidal neurons are extensively connected to each other via recurrent collateral synapses.The nonmyelinated axons of the granule cells, the socalled mossy fibers, show several structural properties that distinguish them from the other synaptic pathways [39]. They project to the CA3 region, mainly traveling within a narrow band termed "stratum lucidum." Several (∼15) large "giant" boutons (∼3-5-μm diameter) emerge from a single mossy fiber axon, either arranged in an en passant manner or attached to the main axon via a short perpendicular axonal branch [1,8,29,31] (Fig. 1c,d). A large mossy fiber bouton typically contacts only a single CA3 pyramidal neuron [21], and a mossy fiber axon contacts a given CA3 pyramidal neuron only once, as boutons are ∼150 μm apart. As the rat hippocampus contains ∼1 million granule cells

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Section: Abstract Presynaptic Recording Mossy Fiber Boutons Mossymentioning

confidence: 99%

Timing and efficacy of transmitter release at mossy fiber synapses in the hippocampal network

Bischofberger

Engel

Frotscher

et al. 2006

Pflugers Arch - Eur J Physiol

115

110

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“…Cerebellar slices were obtained following a previously described technique (Llinas and Sugimori, 1980;Edwards et al, 1989). Briefly, the animals were anesthetized with isoflurane (Isoflurane-Vet, Merial, Italy) and decapitated.…”

Section: Slice Preparation and Patch-clamp Recordingmentioning

confidence: 99%

“…A single slice at a time was placed in the recording chamber and continuously perfused at room temperature (22-25°C) with the saline solution bubbled with the 95% O 2 / 5% CO 2 . A Purkinje cell was visualized using a 40× water-immersion objective of an upright microscope (E600FN, Eclipse, Nikon, Japan), and its upper surface was cleaned by a glass pipette, pulled from sodalime glass to a tip diameter of 10-15 μm, containing the saline solution (Edwards et al, 1989). Pipettes of borosilicate glass, with a tip diameter of 2-3 μm, with resistances between 2.5 and 3.0 MΩ were used for patch-clamp recording.…”

Section: Slice Preparation and Patch-clamp Recordingmentioning

confidence: 99%

Properties and expression of Kv3 channels in cerebellar Purkinje cells

Sacco

Luca

Tempia

2006

Molecular and Cellular Neuroscience

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In cerebellar Purkinje cells, Kv3 potassium channels are indispensable for firing at high frequencies. In Purkinje cells from young mice (P4-P7), Kv3 currents, recorded in whole-cell in slices, activated at −30 mV, with rapid activation and deactivation kinetics, and they were partially blocked by blood depressing substance-I (BDS-I, 1 μM). At positive potentials, Kv3 currents were slowly but completely inactivating, while the recovery from inactivation was about eightfold slower, suggesting that a previous firing activity or a small change of the resting potential could in principle accumulate inactivated Kv3 channels, thereby finely tuning Kv3 current availability for subsequent action potentials. Single-cell RT-PCR analysis showed the expression by all Purkinje cells (n = 10 for each subunit) of Kv3.1, Kv3.3 and Kv3.4 mRNA, while Kv3.2 was not expressed. These results add to the framework for interpreting the physiological function and the molecular determinants of Kv3 currents in cerebellar Purkinje cells.

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“…The overlying tissue was first cleared by applying pressure via a large pipette. 10 Cells were approached with a patch pipette (impedance 2 M ) filled with recording solution (in mM): 100 CsF, 0.5 CaCl 2 , 2 MgCl 2 , 10 HEPES, 10 EGTA, 2 MgATP, 0.1 Na 2 GTP, 20 TEACl, 0.1 leupeptine, 20 phosphocreatine and 50 units/ml phosphocreatine kinase; pH was set to 7.2. Free Ca 2+ concentration was estimated to be smaller than 10 8 M. At these calcium concentrations, the fluoride solution hardly affected the HVA currents in our preparation, as demonstrated previously.…”

Section: In Situ Patch-clampmentioning

confidence: 99%

Calcium currents in pyramidal CA1 neurons in vitro after kindling epileptogenesis in the hippocampus of the rat

1996

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Calcium currents in pyramidal CA1 neurons in vitro after kindling epileptogenesis in the hippocampus of the rat Faas, G.C.; Vreugdenhil, M.; Wadman, W.J. Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. Abstract--Calcium is an important second messenger which plays a role in the regulation of neuronal excitability and in many forms of synaptic plasticity. In kindling epileptogenesis, a model of focal epilepsy, calcium plays an important role. The in situ patch-clamp technique was used to record calcium currents in slices obtained from kindled rats and controls. We found that low-voltage-activated calcium currents, probably of dendritic origin, were larger after kindling (80%). The transient high-voltage-activated calcium currents were also enhanced after kindling (50% higher). The increase of the current is accompanied by a decrease in the time constant of inactivation. The change was still present six weeks after the kindling stimulations were stopped. These data demonstrate that low-voltage-activated calcium currents are involved in epileptogenesis. Their enhancement in the dendrites will boost synaptic depolarization and result in enhanced calcium influx, which is critically dependent on the specific activation pattern.

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A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system

Cited by 1,243 publications

References 23 publications

Timing and efficacy of transmitter release at mossy fiber synapses in the hippocampal network

Timing and efficacy of transmitter release at mossy fiber synapses in the hippocampal network

Properties and expression of Kv3 channels in cerebellar Purkinje cells

Calcium currents in pyramidal CA1 neurons in vitro after kindling epileptogenesis in the hippocampus of the rat

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