Methods for single-cell recording and labeling in vivo

Theta oscillations play a major role in temporarily defining the hippocampal rate code by translating behavioral sequences into neuronal representations. However, mechanisms constraining phase timing and cell-type-specific phase preference are unknown. Here, we employ computational models tuned with evolutionary algorithms to evaluate phase preference of individual CA1 pyramidal cells recorded in mice and rats not engaged in any particular memory task. We applied unbiased and hypothesis-free approaches to identify effects of intrinsic and synaptic factors, as well as cell morphology, in determining phase preference. We found that perisomatic inhibition delivered by complementary populations of basket cells interacts with input pathways to shape phase-locked specificity of deep and superficial pyramidal cells. Somatodendritic integration of fluctuating glutamatergic inputs defined cycle-by-cycle by unsupervised methods demonstrated that firing selection is tuneable across sublayers. Our data identify different mechanisms of phase-locking selectivity that are instrumental for flexible dynamical representations of theta sequences.

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Section: Discussionmentioning

confidence: 95%

“…Single-cell and simultaneous LFP recordings were sampled at 20 kHz/channel with 12 bits precision (Digidata 1440; Molecular Devices). After recording, cells were modulated using the juxtacellular labeling technique with positive current pulses (500-600 ms on-off pulses; 5-18 nA) 68 .…”

Section: Discussionmentioning

confidence: 99%

Multimodal determinants of phase-locked dynamics across deep-superficial hippocampal sublayers during theta oscillations

et al. 2020

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“…Characteristic features such as the laminar profile of theta and sharp-wave ripples, as well as unit activity were used to inform laminar position within the hippocampus. In all cases, electrode position was histologically confirmed 71 Single-cell recordings followed by juxtacellular labeling for post-hoc immune-histochemical identification were obtained in combination with LFP. For juxtacellular recordings, a glass pipette (1.0mm x 0.58mm, ref 601000; A-M Systems) was filled with 1.5-2.5 % Neurobiotin in 0.5 M NaCl (impedance 8-15 MΩ).…”

Section: Electrophysiological Recordingsmentioning

confidence: 99%

Section: Electrophysiological Recordingsmentioning

confidence: 99%

Multimodal determinants of phase-locked dynamics across deep-superficial hippocampal sublayers during theta oscillations

Navas-Olive

Valero

Jurado-Parras

et al. 2020

Preprint

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Theta oscillations play a major role in temporarily defining the hippocampal rate code by translating behavioural sequences into neuronal representations. However, mechanisms constraining phase timing and cell-type specific phase preference are unknown. Here, we employ computational models tuned with evolutionary algorithms to evaluate phase preference of individual CA1 pyramidal cells recorded in mice and rats not engaged in any particular memory task. We applied unbiased and hypothesis-free approaches to identify effects of intrinsic and synaptic factors, as well as cell morphology, in determining phase preference. We found that perisomatic inhibition delivered by complementary populations of basket cells interacts with input pathways to shape phase-locked specificity of deep and superficial pyramidal cells. Somatodendritic integration of fluctuating glutamatergic inputs defined cycle-by-cycle by unsupervised methods demonstrated that firing selection is tuneable across sublayers. Our data identify different mechanisms of phase-locking selectivity that are instrumental for high-level flexible dynamical representations.

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“…These observations raised the possibility that other SOC neurons may fire burst type or regular type patterns. One caveat of loose-seal patch clamp recordings is that cells cannot be identified, unless iontophoretic methods are used to label them (Pinault, 1996; Cid and de la Prida, 2019). As an alternative to identify the morphology and location of recorded cells, whole-cell current clamp recordings were performed in forty ISO anesthetized pups between ages P0 and P11 (Figure 2).…”

Section: Resultsmentioning

confidence: 99%

Characterization of developmental changes in spontaneous electrical activity of superior olivary neurons before hearing onset under injectable or volatile anesthesia

Guilmi

Rodriguez-Contreras

2021

Preprint

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In this work we studied the impact of two widely used anesthetics on the electrical activity of auditory brainstem neurons during development. The spontaneous electrical activity in neonate rats of either sex was analyzed under the injectable mix of ketamine/xylazine (K/X mix) and the volatile anesthetic isoflurane (ISO). We used a ventral craniotomy in mechanically ventilated pups to carry out electrophysiology recordings in the superior olivary complex (SOC) between birth (postnatal day 0, P0) and P12. To characterize neuronal activity of single and ensembles of neurons, we performed patch clamp and multi-electrode experiments under different anesthetic conditions. Our results provide the first study that compares K/X mix and ISO in the same rodent species. We demonstrate that electrical activity of SOC neurons ramps up during development, and that the firing pattern of single units recorded in K/X mix was similar to that reported in ISO anesthetized rat pups. However, ISO displayed a large scatter on its suppressing effects on electrical activity when delivered at 1.5% in the presence or the absence of K/X mix. Taken together, our results shed light on the use of anesthetics for future studies to enable electrophysiology or optical imaging studies in-vivo to obtain functional information on the activity of medial olivochoclear neurons and their role in auditory development.

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Methods for single-cell recording and labeling in vivo

Cited by 16 publications

References 117 publications

Multimodal determinants of phase-locked dynamics across deep-superficial hippocampal sublayers during theta oscillations

Multimodal determinants of phase-locked dynamics across deep-superficial hippocampal sublayers during theta oscillations

Multimodal determinants of phase-locked dynamics across deep-superficial hippocampal sublayers during theta oscillations

Characterization of developmental changes in spontaneous electrical activity of superior olivary neurons before hearing onset under injectable or volatile anesthesia

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