Whole-cell patch-clamp recording and parameters

Kodirov, Sodikdjon A.

doi:10.1007/s12551-023-01055-8

Cited by 10 publications

(5 citation statements)

References 96 publications

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“…We here introduce loose-patch clamp methods ( Stühmer et al, 1983 ) to investigate corresponding effects in native in situ CA1 pyramidal neurons in hippocampal coronal slices for the first time. This approach complements conventional tight patch electrode techniques previously used in both on- and whole-cell electrophysiological single-cell current-clamp ( Liu et al, 2001 ; Minlebaev et al, 2013 ; Ghasemi et al, 2018 ; Vazetdinova et al, 2022 ) or voltage-clamp studies of neuronal cells in hippocampal tissue slices ( Kodirov, 2023 ) or following isolation ( Mathias et al, 1990 ; Klee et al, 1995 ). However, on the one hand, the whole-cell conventional patch clamp configuration permitting membrane current measurement over the entire somal cell membrane involves cell membrane rupture to electrically access the intracellular environment ( French et al, 1990 ).…”

Section: Introductionmentioning

confidence: 90%

Loose-patch clamp analysis applied to voltage-gated ionic currents following pharmacological ryanodine receptor modulation in murine hippocampal cornu ammonis-1 pyramidal neurons

Bertagna,

Ahmad,

Lewis

et al. 2024

Front. Physiol.

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IntroductionThe loose-patch clamp technique was first developed and used in native amphibian skeletal muscle (SkM), offering useful features complementing conventional sharp micro-electrode, gap, or conventional patch voltage clamping. It demonstrated the feedback effects of pharmacological modification of ryanodine receptor (RyR)-mediated Ca2+ release on the Na+ channel (Nav1.4) currents, initiating excitation–contraction coupling in native murine SkM. The effects of the further RyR and Ca2+-ATPase (SERCA) antagonists, dantrolene and cyclopiazonic acid (CPA), additionally implicated background tubular-sarcoplasmic Ca2+ domains in these actions.Materials and methodsWe extend the loose-patch clamp approach to ion current measurements in murine hippocampal brain slice cornu ammonis-1 (CA1) pyramidal neurons. We explored the effects on Na+ currents of pharmacologically manipulating RyR and SERCA-mediated intracellular store Ca2+ release and reuptake. We adopted protocols previously applied to native skeletal muscle. These demonstrated Ca2+-mediated feedback effects on the Na+ channel function.ResultsExperiments applying depolarizing 15 ms duration loose-patch clamp steps to test voltages ranging from −40 to 120 mV positive to the resting membrane potential demonstrated that 0.5 mM caffeine decreased inward current amplitudes, agreeing with the previous SkM findings. It also decreased transient but not prolonged outward current amplitudes. However, 2 mM caffeine affected neither inward nor transient outward but increased prolonged outward currents, in contrast to its increasing inward currents in SkM. Furthermore, similarly and in contrast to previous SkM findings, both dantrolene (10 μM) and CPA (1 μM) pre-administration left both inward and outward currents unchanged. Nevertheless, dantrolene pretreatment still abrogated the effects of subsequent 0.5- and 2-mM caffeine challenges on both inward and outward currents. Finally, CPA abrogated the effects of 0.5 mM caffeine on both inward and outward currents, but with 2 mM caffeine, inward and transient outward currents were unchanged, but sustained outward currents increased.ConclusionWe, thus, extend loose-patch clamping to establish pharmacological properties of murine CA1 pyramidal neurons and their similarities and contrasts with SkM. Here, evoked though not background Ca2+-store release influenced Nav and Kv excitation, consistent with smaller contributions of background store Ca2+ release to resting [Ca2+]. This potential non-canonical mechanism could modulate neuronal membrane excitability or cellular firing rates.

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Section: Introductionmentioning

confidence: 90%

Loose-patch clamp analysis applied to voltage-gated ionic currents following pharmacological ryanodine receptor modulation in murine hippocampal cornu ammonis-1 pyramidal neurons

Bertagna,

Ahmad,

Lewis

et al. 2024

Front. Physiol.

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“…We here introduce loose patch clamp methods to investigate particular features of membrane current in CA1 cells in hippocampal slices for the first time. This complements conventional tight patch electrode techniques proven to be invaluable in both on‐ and whole‐cell electrophysiological single cell current‐clamp 9–12 or voltage‐clamp studies of neuronal cells in hippocampal tissue slices 13 or following isolation 14,15 . The latter techniques utilize electrodes with relatively small tip diameters to achieve required high seal resistances 16,17 .…”

Section: Introductionmentioning

confidence: 94%

Loose patch clamp membrane current measurements in cornus ammonis 1 neurons in murine hippocampal slices

Bertagna,

Ahmad,

Lewis

et al. 2024

Annals of the New York Academy of Sciences

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Hippocampal pyramidal neuronal activity has been previously studied using conventional patch clamp in isolated cells and brain slices. We here introduce the loose patch clamping study of voltage‐activated currents from in situ pyramidal neurons in murine cornus ammonis 1 hippocampal coronal slices. Depolarizing pulses of 15‐ms duration elicited early transient inward, followed by transient and prolonged outward currents in the readily identifiable junctional region between the stratum pyramidalis (SP) and oriens (SO) containing pyramidal cell somas and initial segments. These resembled pyramidal cell currents previously recorded using conventional patch clamp. Shortening the depolarizing pulses to >1–2 ms continued to evoke transient currents; hyperpolarizing pulses to varying voltages evoked decays whose time constants could be shortened to <1 ms, clarifying the speed of clamping in this experimental system. The inward and outward currents had distinct pharmacological characteristics and voltage‐dependent inactivation and recovery from inactivation. Comparative recordings from the SP, known to contain pyramidal cell somas, demonstrated similar current properties. Recordings from the SO and stratum radiatum demonstrated smaller inward and outward current magnitudes and reduced transient outward currents, consistent with previous conventional patch clamp results from their different interneuron types. The loose patch clamp method is thus useful for in situ studies of neurons in hippocampal brain slices.

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“…Rounding out the Issue is a scientific letter contributed by a scientist based in Austria (Kodirov 2023) who discusses different aspects of the patch clamp method-an electrophysiological procedure capable of inferring differences in conduction at the single ion channel level. Starting with a general outline of the technique, this scientific letter relates different aspects of the experimental setup to both expected and anomalous observations at the level of recorded current and electrochemical potential (Kodirov 2023). Having provided this short summary of the current Issue's contents, we round out this Editorial with a short discussion of the pros and cons of writing in both, the formulation of ideas, and in their understanding.…”

Section: Precis Of the Current Issuementioning

confidence: 99%

Biophysical Reviews: And the winner is …

Hall

2023

Biophys Rev

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This Editorial (Vol. 15 Issue 2-Regular Issue) first announces the winner of the 2023 Michèle Auger Award for Young Scientists' Independent Research before then going on to describe the contents of the current Issue. The Editorial closes with a discussion of the pros and cons of writing in the formulation of scientific ideas.Each year, the Biophysical Reviews journal runs a single competition known as the "Michèle Auger Award for Young Scientists' Independent Research". Started in 2020, the award carries out two functions that are important to the journal:(i) The annual award acts as a perpetual memorial for a much admired and well-liked former Editorial Board Member Prof. Michèle Auger (IUPAB 2018), (ii) Helps to recognize the next generation of biophysicists by providing a forum to have their work read and appreciated within the scientific community.In this section, I would like to explain some of the benefits available to those nominated for the award, the process by which this year's award was judged, and finally, to announce the winner of this year's award.

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Whole-cell patch-clamp recording and parameters

Cited by 10 publications

References 96 publications

Loose-patch clamp analysis applied to voltage-gated ionic currents following pharmacological ryanodine receptor modulation in murine hippocampal cornu ammonis-1 pyramidal neurons

Loose-patch clamp analysis applied to voltage-gated ionic currents following pharmacological ryanodine receptor modulation in murine hippocampal cornu ammonis-1 pyramidal neurons

Loose patch clamp membrane current measurements in cornus ammonis 1 neurons in murine hippocampal slices

Biophysical Reviews: And the winner is …

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