Long-lasting single-neuron labeling by in vivo electroporation without microscopic guidance

Oyama, Kei; Ohara, Shinya; Sato, Sho; Karube, Fuyuki; Fujiyama, Fumino; Isomura, Yoshikazu; Mushiake, Hajime; Iijima, Toshio; Tsutsui, Ken

doi:10.1016/j.jneumeth.2013.06.004

Cited by 16 publications

(30 citation statements)

References 17 publications

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“…7). Overall 21 neurons, about 16% of all detection events, were recovered by native fluorescence from 130 penetrations in nine mice, comparable to that of published manual electroporation labelling experiments in vivo 252628 (Fig. 3g).…”

Section: Resultssupporting

confidence: 85%

“…Increased spiking (Fig. 4a2) was observed in this cell post-electroporation25262833. After a 4-day expression, we successfully recovered this neuron's pyramidal morphology at matching location and depth.…”

Section: Resultsmentioning

confidence: 67%

“…However, its applications are restricted to the superficial brain regions accessible to two-photon microscopy. Juxtacellular electroporation and whole-cell (blind) recording, on the other hand, have been manually conducted in deep brain structures but are technically demanding232425262728. Whole-cell experiments also require careful and highly skilled preparation to re-seal the membrane at the end of filling and despite recent efforts, suffer from low yield for delivery of genetic constructs through the patch pipette2930.…”

mentioning

confidence: 99%

“…Fourth, ACE does not require a two-photon microscope, making it cost-effective. Finally, blind patch recording and SCE have been manually conducted in multiple animal models2324252627282930, meaning that ACE can have broad applications in various species in which transgenic animal models are not readily available.…”

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confidence: 99%

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A robot for high yield electrophysiology and morphology of single neurons in vivo

Ouellette

Stoy

et al. 2017

Nat Commun

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Single-cell characterization and perturbation of neurons provides knowledge critical to addressing fundamental neuroscience questions including the structure–function relationship and neuronal cell-type classification. Here we report a robot for efficiently performing in vivo single-cell experiments in deep brain tissues optically difficult to access. This robot automates blind (non-visually guided) single-cell electroporation (SCE) and extracellular electrophysiology, and can be used to characterize neuronal morphological and physiological properties of, and/or manipulate genetic/chemical contents via delivering extraneous materials (for example, genes) into single neurons in vivo. Tested in the mouse brain, our robot successfully reveals the full morphology of single-infragranular neurons recorded in multiple neocortical regions, as well as deep brain structures such as hippocampal CA3, with high efficiency. Our robot thus can greatly facilitate the study of in vivo full morphology and electrophysiology of single neurons in the brain.

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Section: Resultssupporting

confidence: 85%

Section: Resultsmentioning

confidence: 67%

mentioning

confidence: 99%

mentioning

confidence: 99%

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A robot for high yield electrophysiology and morphology of single neurons in vivo

Ouellette

Stoy

et al. 2017

Nat Commun

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“…However, to understand how a neuron relates to a larger neuronal network and how it functions and interacts with other neurons, we need to investigate the precise morphological and histochemical background of the neuron, including its type, which receptors it expresses, and which other neurons it projects to. Staining a single neuron after having recorded from it (Oyama et al 2013) during a behavioral paradigm will allow for such histochemical and morphological investigations and may reveal the functions and relationships of discrete subtypes of striatal neurons that code different reward-related information.…”

Section: Cs Tonic Cs Phasicmentioning

confidence: 99%

Discrete coding of stimulus value, reward expectation, and reward prediction error in the dorsal striatum

Oyama

Tateyama

Hernádi

et al. 2015

Journal of Neurophysiology

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To investigate how the striatum integrates sensory information with reward information for behavioral guidance, we recorded single-unit activity in the dorsal striatum of head-fixed rats participating in a probabilistic Pavlovian conditioning task with auditory conditioned stimuli (CSs) in which reward probability was fixed for each CS but parametrically varied across CSs. We found that the activity of many neurons was linearly correlated with the reward probability indicated by the CSs. The recorded neurons could be classified according to their firing patterns into functional subtypes coding reward probability in different forms such as stimulus value, reward expectation, and reward prediction error. These results suggest that several functional subgroups of dorsal striatal neurons represent different kinds of information formed through extensive prior exposure to CS-reward contingencies.

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In vivo electroporation to physiologically identified deep brain regions in postnatal mammals

et al. 2014

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Genetic manipulation is widely used to research the central nervous system (CNS). The manipulation of molecular expression in a small number of neurons permits the detailed investigation of the role of specific molecules on the function and morphology of the neurons. Electroporation is a broadly used technique for gene transfer in the CNS. However, the targeting of gene transfer using electroporation in postnatal animals was restricted to the cortex, hippocampus, or the region facing the ventricle in previous reports. Electroporation targeting of deep brain structures, such as the thalamus, has been difficult. We introduce a novel electroporation technique that enables gene transfer to a physiologically identified deep brain region using a glass pipette. We recorded neural activity in young-adult mice to identify the location of the lateral geniculate nucleus (LGN) of the thalamus, using a glass pipette electrode containing the plasmid DNA encoding enhanced green fluorescent protein (EGFP). The location of the LGN was confirmed by monitoring visual responses, and the plasmid solution was pressure-injected into the recording site. Voltage pulses were delivered through the glass pipette electrode. Several EGFP-labeled somata and dendrites were observed in the LGN after a few weeks, and labeled axons were found in the visual cortex. The EGFP-expressing structures were observed in detail sufficient to reconstruct their morphology in three dimensions. We further confirmed the applicability of this technique in cats. This method should be useful for the transfer of various genes into cells in physiologically identified brain regions in rodents and gyrencephalic mammals.

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Long-lasting single-neuron labeling by in vivo electroporation without microscopic guidance

Cited by 16 publications

References 17 publications

A robot for high yield electrophysiology and morphology of single neurons in vivo

A robot for high yield electrophysiology and morphology of single neurons in vivo

Discrete coding of stimulus value, reward expectation, and reward prediction error in the dorsal striatum

In vivo electroporation to physiologically identified deep brain regions in postnatal mammals

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