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Article ; Online: Advances in the automation of whole-cell patch clamp technology.

Suk, Ho-Jun / Boyden, Edward S / van Welie, Ingrid

2019 Volume 326, Page(s) 108357

Abstract: Electrophysiology is the study of neural activity in the form of local field potentials, current flow through ion channels, calcium spikes, back propagating action potentials and somatic action potentials, all measurable on a millisecond timescale. ... ...

Abstract	Electrophysiology is the study of neural activity in the form of local field potentials, current flow through ion channels, calcium spikes, back propagating action potentials and somatic action potentials, all measurable on a millisecond timescale. Despite great progress in imaging technologies and sensor proteins, none of the currently available tools allow imaging of neural activity on a millisecond timescale and beyond the first few hundreds of microns inside the brain. The patch clamp technique has been an invaluable tool since its inception several decades ago and has generated a wealth of knowledge about the nature of voltage- and ligand-gated ion channels, sub-threshold and supra-threshold activity, and characteristics of action potentials related to higher order functions. Many techniques that evolve to be standardized tools in the biological sciences go through a period of transformation in which they become, at least to some degree, automated, in order to improve reproducibility, throughput and standardization. The patch clamp technique is currently undergoing this transition, and in this review, we will discuss various aspects of this transition, covering advances in automated patch clamp technology both in vitro and in vivo.
MeSH term(s)	Animals ; Automation, Laboratory/methods ; Electrophysiology/methods ; Electrophysiology/trends ; Humans ; Neurosciences/methods ; Neurosciences/trends ; Patch-Clamp Techniques/methods ; Patch-Clamp Techniques/trends
Language	English
Publishing date	2019-07-20
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	282721-9
ISSN	1872-678X ; 0165-0270
ISSN (online)	1872-678X
ISSN	0165-0270
DOI	10.1016/j.jneumeth.2019.108357
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Article ; Online: Closed-Loop Real-Time Imaging Enables Fully Automated Cell-Targeted Patch-Clamp Neural Recording In Vivo.

Suk, Ho-Jun / van Welie, Ingrid / Kodandaramaiah, Suhasa B / Allen, Brian / Forest, Craig R / Boyden, Edward S

Neuron

2017 Volume 96, Issue 1, Page(s) 244–245

Language	English
Publishing date	2017-09-27
Publishing country	United States
Document type	Journal Article ; Published Erratum
ZDB-ID	808167-0
ISSN	1097-4199 ; 0896-6273
ISSN (online)	1097-4199
ISSN	0896-6273
DOI	10.1016/j.neuron.2017.09.012
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Zs.A 2496: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Conditional Spike Transmission Mediated by Electrical Coupling Ensures Millisecond Precision-Correlated Activity among Interneurons In Vivo.

van Welie, Ingrid / Roth, Arnd / Ho, Sara S N / Komai, Shoji / Häusser, Michael

Neuron

2016 Volume 90, Issue 4, Page(s) 810–823

Abstract: Many GABAergic interneurons are electrically coupled and in vitro can display correlated activity with millisecond precision. However, the mechanisms underlying correlated activity between interneurons in vivo are unknown. Using dual patch-clamp ... ...

Abstract	Many GABAergic interneurons are electrically coupled and in vitro can display correlated activity with millisecond precision. However, the mechanisms underlying correlated activity between interneurons in vivo are unknown. Using dual patch-clamp recordings in vivo, we reveal that in the presence of spontaneous background synaptic activity, electrically coupled cerebellar Golgi cells exhibit robust millisecond precision-correlated activity which is enhanced by sensory stimulation. This precisely correlated activity results from the cooperative action of two mechanisms. First, electrical coupling ensures slow subthreshold membrane potential correlations by equalizing membrane potential fluctuations, such that coupled neurons tend to approach action potential threshold together. Second, fast spike-triggered spikelets transmitted through gap junctions conditionally trigger postjunctional spikes, depending on both neurons being close to threshold. Electrical coupling therefore controls the temporal precision and degree of both spontaneous and sensory-evoked correlated activity between interneurons, by the cooperative effects of shared synaptic depolarization and spikelet transmission.
MeSH term(s)	Action Potentials/physiology ; Animals ; Electric Stimulation/methods ; Gap Junctions/physiology ; Interneurons/physiology ; Membrane Potentials/physiology ; Mice ; Neural Inhibition/physiology ; Patch-Clamp Techniques/methods ; Synapses/physiology ; Synaptic Transmission/physiology
Language	English
Publishing date	2016-05-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	808167-0
ISSN	1097-4199 ; 0896-6273
ISSN (online)	1097-4199
ISSN	0896-6273
DOI	10.1016/j.neuron.2016.04.013
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Zs.A 2496: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 92: Show issues

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Article ; Online: The metamorphosis of the developing cerebellar microcircuit.

van Welie, Ingrid / Smith, Ikuko T / Watt, Alanna J

Current opinion in neurobiology

2011 Volume 21, Issue 2, Page(s) 245–253

Abstract: The cerebellar cortical circuit with its organized and repetitive structure provides an excellent model system for studying how brain circuits are formed during development. The emergence of the mature brain requires that appropriate synaptic connections ...

Abstract	The cerebellar cortical circuit with its organized and repetitive structure provides an excellent model system for studying how brain circuits are formed during development. The emergence of the mature brain requires that appropriate synaptic connections are formed and refined, which in the rodent cerebellum occurs primarily during the first three postnatal weeks. Developing circuits typically differ substantially from their mature counterparts, which suggests that development may not simply involve synaptic refinement, but rather involves restructuring of key synaptic components and network connections, in a manner reminiscent of metamorphosis. Here, we discuss recent evidence that, taken together, suggests that transient features of developing cerebellar synapses may act to coordinate network activity, and thereby shape the development of the cerebellar microcircuit.
MeSH term(s)	Animals ; Cerebellum/growth & development ; Cerebellum/ultrastructure ; Humans ; Neural Pathways/growth & development ; Neural Pathways/ultrastructure ; Neurogenesis/physiology ; Synapses/physiology ; Synapses/ultrastructure
Language	English
Publishing date	2011-02-24
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1078046-4
ISSN	1873-6882 ; 0959-4388
ISSN (online)	1873-6882
ISSN	0959-4388
DOI	10.1016/j.conb.2011.01.009
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Zs.A 3260: Show issues

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Article ; Online: Closed-Loop Real-Time Imaging Enables Fully Automated Cell-Targeted Patch-Clamp Neural Recording In Vivo.

Suk, Ho-Jun / van Welie, Ingrid / Kodandaramaiah, Suhasa B / Allen, Brian / Forest, Craig R / Boyden, Edward S

Neuron

2017 Volume 95, Issue 5, Page(s) 1037–1047.e11

Abstract: Targeted patch-clamp recording is a powerful method for characterizing visually identified cells in intact neural circuits, but it requires skill to perform. We previously developed an algorithm that automates "blind" patching in vivo, but full ... ...

Abstract	Targeted patch-clamp recording is a powerful method for characterizing visually identified cells in intact neural circuits, but it requires skill to perform. We previously developed an algorithm that automates "blind" patching in vivo, but full automation of visually guided, targeted in vivo patching has not been demonstrated, with currently available approaches requiring human intervention to compensate for cell movement as a patch pipette approaches a targeted neuron. Here we present a closed-loop real-time imaging strategy that automatically compensates for cell movement by tracking cell position and adjusting pipette motion while approaching a target. We demonstrate our system's ability to adaptively patch, under continuous two-photon imaging and real-time analysis, fluorophore-expressing neurons of multiple types in the living mouse cortex, without human intervention, with yields comparable to skilled human experimenters. Our "imagepatching" robot is easy to implement and will help enable scalable characterization of identified cell types in intact neural circuits.
MeSH term(s)	Animals ; Automation/methods ; Computer Systems ; Fluorescent Dyes ; Functional Neuroimaging/methods ; Mice ; Neurons/physiology ; Patch-Clamp Techniques/methods
Chemical Substances	Fluorescent Dyes
Language	English
Publishing date	2017-09-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	808167-0
ISSN	1097-4199 ; 0896-6273
ISSN (online)	1097-4199
ISSN	0896-6273
DOI	10.1016/j.neuron.2017.08.011
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Zs.A 2496: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Bidirectional control of BK channel open probability by CAMKII and PKC in medial vestibular nucleus neurons.

van Welie, Ingrid / du Lac, Sascha

Journal of neurophysiology

2011 Volume 105, Issue 4, Page(s) 1651–1659

Abstract: Large conductance K(+) (BK) channels are a key determinant of neuronal excitability. Medial vestibular nucleus (MVN) neurons regulate eye movements to ensure image stabilization during head movement, and changes in their intrinsic excitability may play a ...

Abstract	Large conductance K(+) (BK) channels are a key determinant of neuronal excitability. Medial vestibular nucleus (MVN) neurons regulate eye movements to ensure image stabilization during head movement, and changes in their intrinsic excitability may play a critical role in plasticity of the vestibulo-ocular reflex. Plasticity of intrinsic excitability in MVN neurons is mediated by kinases, and BK channels influence excitability, but whether endogenous BK channels are directly modulated by kinases is unknown. Double somatic patch-clamp recordings from MVN neurons revealed large conductance potassium channel openings during spontaneous action potential firing. These channels displayed Ca(2+) and voltage dependence in excised patches, identifying them as BK channels. Recording isolated single channel currents at physiological temperature revealed a novel kinase-mediated bidirectional control in the range of voltages over which BK channels are activated. Application of activated Ca(2+)/calmodulin-dependent kinase II (CAMKII) increased BK channel open probability by shifting the voltage activation range towards more hyperpolarized potentials. An opposite shift in BK channel open probability was revealed by inhibition of phosphatases and was occluded by blockade of protein kinase C (PKC), suggesting that active PKC associated with BK channel complexes in patches was responsible for this effect. Accordingly, direct activation of endogenous PKC by PMA induced a decrease in BK open probability. BK channel activity affects excitability in MVN neurons and bidirectional control of BK channels by CAMKII, and PKC suggests that cellular signaling cascades engaged during plasticity may dynamically control excitability by regulating BK channel open probability.
MeSH term(s)	Action Potentials/physiology ; Animals ; Body Temperature/physiology ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/physiology ; Female ; Large-Conductance Calcium-Activated Potassium Channels/physiology ; Male ; Mice ; Mice, Inbred C57BL ; Models, Animal ; Neuronal Plasticity/physiology ; Neurons/cytology ; Neurons/physiology ; Patch-Clamp Techniques ; Protein Kinase C/physiology ; Vestibular Nuclei/physiology
Chemical Substances	Large-Conductance Calcium-Activated Potassium Channels ; Protein Kinase C (EC 2.7.11.13) ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 (EC 2.7.11.17)
Language	English
Publishing date	2011-02-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	80161-6
ISSN	1522-1598 ; 0022-3077
ISSN (online)	1522-1598
ISSN	0022-3077
DOI	10.1152/jn.00058.2011
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Article: Background activity regulates excitability of rat hippocampal CA1 pyramidal neurons by adaptation of a K+ conductance.

van Welie, Ingrid / van Hooft, Johannes A / Wadman, Wytse J

Journal of neurophysiology

2006 Volume 95, Issue 3, Page(s) 2007–2012

Abstract: In the in vivo brain background synaptic activity has a strong modulatory influence on neuronal excitability. Here we report that in rat hippocampal slices, blockade of endogenous in vitro background activity results in an increased excitability of CA1 ... ...

Abstract	In the in vivo brain background synaptic activity has a strong modulatory influence on neuronal excitability. Here we report that in rat hippocampal slices, blockade of endogenous in vitro background activity results in an increased excitability of CA1 pyramidal neurons within tens of minutes. The increase in excitability constitutes a leftward shift in the input-output relationship of pyramidal neurons, indicating a reduced threshold for the induction of action potentials. The increase in excitability results from an adaptive decrease in a sustained K+ conductance, as recorded from somatic cell-attached patches. After 20 min of blockade of background activity, the mean sustained K+ current amplitude in somatic patches was reduced to 46 +/- 9% of that in time-matched control patches. Blockade of background activity did not affect fast Na+ conductance. Together, these results suggests that the reduction in K+ conductance serves as an adaptive mechanism to increase the excitability of CA1 pyramidal neurons in response to changes in background activity such that the dynamic range of the input-output relationship is effectively maintained.
MeSH term(s)	Action Potentials/physiology ; Adaptation, Physiological/physiology ; Animals ; Biological Clocks/physiology ; Cells, Cultured ; Electric Conductivity ; Male ; Neuronal Plasticity/physiology ; Potassium/metabolism ; Pyramidal Cells/physiology ; Rats ; Rats, Wistar ; Synaptic Transmission/physiology
Chemical Substances	Potassium (RWP5GA015D)
Language	English
Publishing date	2006-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80161-6
ISSN	1522-1598 ; 0022-3077
ISSN (online)	1522-1598
ISSN	0022-3077
DOI	10.1152/jn.00220.2005
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Article: Low affinity block of native and cloned hyperpolarization-activated Ih channels by Ba2+ ions.

van Welie, Ingrid / Wadman, Wytse J / van Hooft, Johannes A

European journal of pharmacology

2005 Volume 507, Issue 1-3, Page(s) 15–20

Abstract: Ba2+ is commonly used to discriminate two classes of ion currents. The classical inward-rectifying K+ current, I(Kir), is blocked by low millimolar concentrations of Ba2+, whereas the hyperpolarization-activated cation current, I(h), is assumed not to be ...

Abstract	Ba2+ is commonly used to discriminate two classes of ion currents. The classical inward-rectifying K+ current, I(Kir), is blocked by low millimolar concentrations of Ba2+, whereas the hyperpolarization-activated cation current, I(h), is assumed not to be sensitive to Ba2+. Here we investigated the effects of Ba2+ on I(h) currents recorded from rat hippocampal CA1 pyramidal neurons, and on cloned I(h) channels composed of either HCN1 or HCN2 subunits transiently expressed in Human Embryonic Kidney (HEK) 293 cells. The results show that low millimolar concentrations of Ba2+ reduce the maximal I(h) conductance (IC50 approximately 3-5 mM) in both CA1 pyramidal neurons and in HEK 293 cells without specificity for HCN1 or HCN2 subunits. In addition, Ba2+ decreases the rate of activation and increases the rate of deactivation of I(h) currents. Neither the half-maximal voltage of activation, V(h), nor the reversal potential of the I(h) channels were affected by Ba2+. The combined results suggest that B2+, at concentrations commonly used to block I(Kir) currents, also reduces the conductance of I(h) channels without subunit specificity, and affects the kinetics of I(h) channel gating.
MeSH term(s)	Animals ; Barium/pharmacology ; Cell Line ; Cloning, Molecular/methods ; Cyclic Nucleotide-Gated Cation Channels ; Humans ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Ion Channels/antagonists & inhibitors ; Ion Channels/physiology ; Male ; Membrane Potentials/drug effects ; Membrane Potentials/physiology ; Potassium Channels ; Rats ; Rats, Wistar
Chemical Substances	Cyclic Nucleotide-Gated Cation Channels ; HCN1 protein, human ; HCN2 protein, human ; Hcn1 protein, rat ; Hcn2 protein, rat ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Ion Channels ; Potassium Channels ; Barium (24GP945V5T)
Language	English
Publishing date	2005-01-10
Publishing country	Netherlands
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80121-5
ISSN	1879-0712 ; 0014-2999
ISSN (online)	1879-0712
ISSN	0014-2999
DOI	10.1016/j.ejphar.2004.11.051
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Zs.A 522: Show issues

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Article: Homeostatic scaling of neuronal excitability by synaptic modulation of somatic hyperpolarization-activated Ih channels.

van Welie, Ingrid / van Hooft, Johannes A / Wadman, Wytse J

Proceedings of the National Academy of Sciences of the United States of America

2004 Volume 101, Issue 14, Page(s) 5123–5128

Abstract: The hyperpolarization-activated cation current (Ih) plays an important role in determining membrane potential and firing characteristics of neurons and therefore is a potential target for regulation of intrinsic excitability. Here we show that an ... ...

Abstract	The hyperpolarization-activated cation current (Ih) plays an important role in determining membrane potential and firing characteristics of neurons and therefore is a potential target for regulation of intrinsic excitability. Here we show that an increase in AMPA-receptor-dependent synaptic activity induced by alpha-latrotoxin or glutamate application as well as direct depolarization results in an increase in Ih recorded from cell-attached patches in hippocampal CA1 pyramidal neurons. This mechanism requires Ca2+ influx but not increased levels of cAMP. Artificially increasing Ih by using a dynamic clamp during whole-cell current clamp recordings results in reduced firing rates in response to depolarizing current injections. We conclude that modulation of somatic Ih represents a previously uncharacterized mechanism of homeostatic plasticity, allowing a neuron to control its excitability in response to changes in synaptic activity on a relatively short-term time scale.
MeSH term(s)	Animals ; Glutamic Acid/physiology ; Homeostasis ; Ion Channels/physiology ; Male ; Neurons/physiology ; Rats ; Rats, Wistar ; Receptors, AMPA/physiology ; Synapses/physiology ; gamma-Aminobutyric Acid/physiology
Chemical Substances	Ion Channels ; Receptors, AMPA ; Glutamic Acid (3KX376GY7L) ; gamma-Aminobutyric Acid (56-12-2)
Language	English
Publishing date	2004-03-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.0307711101
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Article: Different levels of Ih determine distinct temporal integration in bursting and regular-spiking neurons in rat subiculum.

van Welie, Ingrid / Remme, Michiel W H / van Hooft, Johannes A / Wadman, Wytse J

The Journal of physiology

2006 Volume 576, Issue Pt 1, Page(s) 203–214

Abstract: Pyramidal neurons in the subiculum typically display either bursting or regular-spiking behaviour. Although this classification into two neuronal classes is well described, it is unknown how these two classes of neurons contribute to the integration of ... ...

Abstract	Pyramidal neurons in the subiculum typically display either bursting or regular-spiking behaviour. Although this classification into two neuronal classes is well described, it is unknown how these two classes of neurons contribute to the integration of input to the subiculum. Here, we report that bursting neurons possess a hyperpolarization-activated cation current (I(h)) that is two-fold larger (conductance, 5.3 +/- 0.5 nS) than in regular-spiking neurons (2.2 +/- 0.6 nS), whereas I(h) exhibits similar voltage-dependent and kinetic properties in both classes of neurons. Bursting and regular-spiking neurons display similar morphology. The difference in I(h) between the two classes of neurons is not responsible for the distinct firing patterns, as neither pharmacological blockade of I(h) nor enhancement of I(h) using a dynamic clamp affects the qualitative firing patterns. Instead, the difference in I(h) between bursting and regular-spiking neurons determines the temporal integration of evoked synaptic input from the CA1 area. In response to stimulation at 50 Hz, bursting neurons, with a large I(h), show approximately 50% less temporal summation than regular-spiking neurons. The amount of temporal summation in both neuronal classes is equal after pharmacological blockade of I(h). A computer simulation model of a subicular neuron with the properties of either a bursting or a regular-spiking neuron confirmed the pivotal role of I(h) in temporal integration of synaptic input. These data suggest that in the subicular network, bursting neurons are better suited to discriminate the content of high-frequency input, such as that occurring during gamma oscillations, than regular-spiking neurons.
MeSH term(s)	Action Potentials/physiology ; Animals ; Cations ; Computer Simulation ; Electrophysiology ; Hippocampus/physiology ; Ion Channels/physiology ; Male ; Mathematics ; Membrane Potentials/physiology ; Neurons/classification ; Neurons/physiology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Synapses/physiology
Chemical Substances	Cations ; Ion Channels
Language	English
Publishing date	2006-06-29
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3115-x
ISSN	1469-7793 ; 0022-3751
ISSN (online)	1469-7793
ISSN	0022-3751
DOI	10.1113/jphysiol.2006.113944
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