How cesium dialysis affects the passive properties of pyramidal neurons: implications for voltage clamp studies of persistent sodium current

Fleidervish, Ilya A.; Libman, Lior

doi:10.1088/1367-2630/10/3/035001

Cited by 8 publications

(5 citation statements)

References 29 publications

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“…Although our voltage clamp could have been arguably improved using internal cesium chloride in the intracellular pipette solution, we found that replacing internal K ϩ with cesium led to membrane resting values near Ϫ10 mV, which made voltage clamping cells at voltages between Ϫ95 mV and Ϫ45 mV more difficult due to the substantial increase in mean current required to clamp cells near these voltages. This is expected and results from reducing internal K ϩ concentration with cesium and the resulting change in the K ϩ reversal potential (Fleidervish and Libman, 2008). Furthermore, in our experience, as well as others (Kato et al, 2017), the use of QX-314 in the pipette solution significantly reduces synaptic-mediated spontaneous membrane voltage fluctuations during in vivo patch recordings.…”

Section: Potential Issues With Voltage-clamp and Dendritic Contributisupporting

confidence: 63%

Voltage-Dependent Membrane Properties Shape the Size But Not the Frequency Content of Spontaneous Voltage Fluctuations in Layer 2/3 Somatosensory Cortex

2019

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Under awake and idling conditions, spontaneous intracellular membrane voltage is characterized by large, synchronous, low-frequency fluctuations. Although these properties reflect correlations in synaptic inputs, intrinsic membrane properties often indicate voltagedependent changes in membrane resistance and time constant values that can amplify and help to generate low-frequency voltage fluctuations. The specific contribution of intrinsic and synaptic factors to the generation of spontaneous fluctuations, however, remains poorly understood. Using visually guided intracellular recordings of somatosensory layer 2/3 pyramidal cells and interneurons in awake male and female mice, we measured the spectrum and size of voltage fluctuation and intrinsic cellular properties at different voltages. In both cell types, depolarizing neurons increased the size of voltage fluctuations. Amplitude changes scaled with voltage-dependent changes in membrane input resistance. Because of the small membrane time constants observed in both pyramidal cells and interneuron cell bodies, the low-frequency content of membrane fluctuations reflects correlations in the synaptic current inputs rather than significant filtering associated with membrane capacitance. Further, blocking synaptic inputs minimally altered somatic membrane resistance and time constant values. Overall, these results indicate that spontaneous synaptic inputs generate a low-conductance state in which the amplitude, but not frequency structure, is influenced by intrinsic membrane properties.In the absence of sensory drive, cortical activity in awake animals is associated with self-generated and seemingly random membrane voltage fluctuations characterized by large amplitude and low frequency. Partially, these properties reflect correlations in synaptic input. Nonetheless, neurons express voltage-dependent intrinsic properties that can potentially influence the amplitude and frequency of spontaneous activity. Using visually guided intracellular recordings of cortical neurons in awake mice, we measured the voltage dependence of spontaneous voltage fluctuations and intrinsic membrane properties. We show that voltagedependent changes in membrane resistance amplify synaptic activity, whereas the frequency of voltage fluctuations reflects correlations in synaptic inputs. Last, synaptic activity has a small impact on intrinsic membrane properties in both pyramidal cells and interneurons.

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Section: Potential Issues With Voltage-clamp and Dendritic Contributisupporting

confidence: 63%

Voltage-Dependent Membrane Properties Shape the Size But Not the Frequency Content of Spontaneous Voltage Fluctuations in Layer 2/3 Somatosensory Cortex

2019

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“…We therefore sought to replicate previous studies and recorded mEPSCs in hippocampal neurons electrophysiologically. Recordings were performed under voltage-clamp conditions, with the cell held at −70 mV, taking steps to maximize the sensitivity of mEPSC detection, including the use of a high impedance (low noise) amplifier and a cesium-containing internal solution to increase input resistance (Spruston et al, 1993 ; Fleidervish and Libman, 2008 ). In contrast to the increase in mini frequency we observed using direct optical measurements, we found a small reduction in mEPSC frequency ( Figures 4A,B ).…”

Section: Resultsmentioning

confidence: 99%

A Novel Optical Quantal Analysis of Miniature Events Reveals Enhanced Frequency Following Amyloid β Exposure

Taylor

Tong

Jeans

et al. 2020

Front. Cell. Neurosci.

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Non-evoked miniature release of neurotransmitters is increasingly recognized as playing an important role in neural function and is implicated in synaptic plasticity, metaplasticity, and homeostasis. Spontaneous miniature release events (minis) are usually measured electrophysiologically by recording the miniature postsynaptic currents (mEPSCs) that they evoke. However, this indirect technique can be confounded by changes within the postsynaptic neuron. Here, using the fluorescent probe SynaptopHluorin 2×, we have developed an optical method for the measurement of minis that enables direct assessment of release events. We use the technique to reveal that the frequency of minis following incubation of hippocampal neurons with Amyloid β oligomers (Aβo) is increased. Electrophysiological mEPSC recordings obtained under the same conditions report a decrease in frequency, with the discrepancy likely due to Aβo-induced changes in quantal size. Optical quantal analysis of minis may therefore have a role in the study of minis in both normal physiology and disease, as it can circumvent potential confounds caused by postsynaptic changes.

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“…The pH of the pipette solution was adjusted to 7.3 with KOH and the osmolality was set to 295 mOsm. Potassium-methylsulfate as internal solution provided a physiological space clamp ( Fleidervish and Libman, 2008 ). Dual patch-clamp whole-cell recordings (EPC9/dual, HEKA, Germany) were made in the frontal cortex (medial agranular and anterior cingulate cortex) with the use of × 40 water-immersion objective (Axioskop FS, Carl Zeiss, Germany).…”

Section: Methodsmentioning

confidence: 99%

Functional effects of distinct innervation styles of pyramidal cells by fast spiking cortical interneurons

Kubota

Kondo

Nomura

et al. 2015

eLife

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Inhibitory interneurons target precise membrane regions on pyramidal cells, but differences in their functional effects on somata, dendrites and spines remain unclear. We analyzed inhibitory synaptic events induced by cortical, fast-spiking (FS) basket cells which innervate dendritic shafts and spines as well as pyramidal cell somata. Serial electron micrograph (EMg) reconstructions showed that somatic synapses were larger than dendritic contacts. Simulations with precise anatomical and physiological data reveal functional differences between different innervation styles. FS cell soma-targeting synapses initiate a strong, global inhibition, those on shafts inhibit more restricted dendritic zones, while synapses on spines may mediate a strictly local veto. Thus, FS cell synapses of different sizes and sites provide functionally diverse forms of pyramidal cell inhibition.DOI: http://dx.doi.org/10.7554/eLife.07919.001

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How cesium dialysis affects the passive properties of pyramidal neurons: implications for voltage clamp studies of persistent sodium current

Cited by 8 publications

References 29 publications

Voltage-Dependent Membrane Properties Shape the Size But Not the Frequency Content of Spontaneous Voltage Fluctuations in Layer 2/3 Somatosensory Cortex

Voltage-Dependent Membrane Properties Shape the Size But Not the Frequency Content of Spontaneous Voltage Fluctuations in Layer 2/3 Somatosensory Cortex

A Novel Optical Quantal Analysis of Miniature Events Reveals Enhanced Frequency Following Amyloid β Exposure

Functional effects of distinct innervation styles of pyramidal cells by fast spiking cortical interneurons

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