High-throughput cellular-resolution synaptic connectivity mapping in vivo with concurrent two-photon optogenetics and volumetric Ca2+ imaging

AbstractThe ability to measure synaptic connectivity and properties is essential for understanding neuronal circuits. However, existing methods that allow such measurements at cellular resolution are laborious and technically demanding. Here, we describe a system that allows such measurements in a high-throughput way by combining two-photon optogenetics and volumetric Ca2+ imaging with whole-cell recording. We reveal a circuit motif for generating fast undulatory locomotion in zebrafish.

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Compensation of physiological motion enables high-yield whole-cell recording in vivo

10.1101/2020.06.09.143008 ◽

2020 ◽

Author(s):

William M. Stoy ◽

Bo Yang ◽

Ali Kight ◽

Nathaniel C. Wright ◽

Peter Y. Borden ◽

...

Keyword(s):

Single Cells ◽

Strong Dependence ◽

High Yield ◽

Patch Clamp Recording ◽

Gold Standard Method ◽

Whole Cell ◽

Whole Cell Recording ◽

Cell Recording ◽

Living Mouse

1.1.1AbstractWhole-cell patch-clamp recording in vivo is the gold-standard method for measuring subthreshold electrophysiology from single cells during behavioural tasks, sensory stimulations, and optogenetic manipulation. However, these recordings require a tight, gigaohm resistance, seal between a glass pipette electrode’s aperture and a cell’s membrane. These seals are difficult to form, especially in vivo, in part because of a strong dependence on the distance between the pipette aperture and cell membrane. We elucidate and utilize this dependency to develop an autonomous method for placement and synchronization of pipette’s tip aperture to the membrane of a nearby, moving neuron, which enables high-yield seal formation and subsequent recordings in the deep in the brain of the living mouse, in the thalamus. This synchronization procedure nearly doubles the reported gigaseal yield in the thalamus (>3 mm below the pial surface) from 26% (n=17/64) to 48% (n=32/66). Whole-cell recording yield improved from 10% (n = 9/88) to 24% (n=18/76) when motion compensation was used during the gigaseal formation. As an example of its application, we utilized this system to investigate the role of the sensory environment and ventral posterior medial region (VPM) projection synchrony on intracellular dynamics in the barrel cortex. This method results in substantially greater subcortical whole-cell recording yield than previously reported and thus makes pan-brain whole-cell electrophysiology practical in the living mouse brain.

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Faculty Opinions recommendation of Transfection via whole-cell recording in vivo: bridging single-cell physiology, genetics and connectomics.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.10314962.11583054 ◽

2011 ◽

Author(s):

Nathan Dascal

Keyword(s):

Single Cell ◽

Cell Physiology ◽

Whole Cell ◽

Whole Cell Recording ◽

Cell Recording ◽

Single Cell Physiology

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Faculty Opinions recommendation of Transfection via whole-cell recording in vivo: bridging single-cell physiology, genetics and connectomics.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.10314962.11124060 ◽

2011 ◽

Author(s):

Yi Eve Sun ◽

Weihong Ge

Keyword(s):

Single Cell ◽

Cell Physiology ◽

Whole Cell ◽

Whole Cell Recording ◽

Cell Recording ◽

Single Cell Physiology

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Whole-Cell Recording and Voltage-Sensitive Dye Imaging In Vivo

Cold Spring Harbor Protocols ◽

10.1101/pdb.prot5232 ◽

2009 ◽

Vol 2009 (6) ◽

pp. pdb.prot5232-pdb.prot5232 ◽

Cited By ~ 1

Author(s):

C. Petersen

Keyword(s):

Voltage Sensitive Dye ◽

Whole Cell ◽

Whole Cell Recording ◽

Cell Recording ◽

Voltage Sensitive Dye Imaging

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High-velocity stimulation evokes “dense” population response in layer 2/3 vibrissal cortex

Journal of Neurophysiology ◽

10.1152/jn.00815.2016 ◽

2017 ◽

Vol 117 (3) ◽

pp. 1218-1228 ◽

Cited By ~ 9

Author(s):

Yadollah Ranjbar-Slamloo ◽

Ehsan Arabzadeh

Keyword(s):

High Velocity ◽

Sensory Cortex ◽

Dense Population ◽

Population Response ◽

Whole Cell ◽

Two Photon ◽

Whole Cell Recording ◽

Layer 2 ◽

Cell Recording ◽

Standard Range

Supragranular layers of sensory cortex are known to exhibit sparse firing. In rodent vibrissal cortex, a small fraction of neurons in layer 2 and 3 (L2/3) respond to whisker stimulation. In this study, we combined whole cell recording and two-photon imaging in anesthetized mice and quantified the synaptic response and spiking profile of L2/3 neurons. Previous literature has shown that neurons across layers of vibrissal cortex are tuned to the velocity of whisker movement. We therefore used a broad range of stimuli that included the standard range of velocities (0–1.2 deg/ms) and extended to a “sharp” high-velocity deflection (3.8 deg/ms). Consistent with previous literature, whole cell recording revealed a sparse response to the standard range of velocities: although all recorded cells showed tuning to velocity in their postsynaptic potentials, only a small fraction produced stimulus-evoked spikes. In contrast, the sharp stimulus evoked reliable spiking in the majority of neurons. The action potential threshold of spikes evoked by the sharp stimulus was significantly lower than that of the spontaneous spikes. Juxtacellular recordings confirmed that application of sharp stimulus to single or multiple whiskers produced temporally precise spiking with minimal trial-to-trial spike count variability (Fano factors equal or close to the theoretical minimum). Two-photon imaging further confirmed that most neurons that were not responsive to the standard deflections responded to the sharp stimulus. Altogether, our results indicate that sparseness in L2/3 cortex depends on the choice of stimulus: strong single- or multiwhisker stimulation can induce the transition from sparse to “dense” population response. NEW & NOTEWORTHY In superficial layers of sensory cortex, only a small fraction of neurons fire most of the spontaneous and sensory evoked spikes. However, the functional relevance of such “sparse” activity remains unknown. We found that a “dense” population response is evoked by high-velocity micromotions applied to whiskers. Our results suggest that flashes of precisely timed population response on an almost silent background can provide a high capacity for coding of ecologically salient stimuli.

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