Feedforward Inhibitory Connections From Multiple Thalamic Cells to Multiple Regular-Spiking Cells in Layer 4 of the Somatosensory Cortex

2006 ◽  
Vol 96 (4) ◽  
pp. 1746-1754 ◽  
Author(s):  
Tsuyoshi Inoue ◽  
Keiji Imoto
Keyword(s):  
Patch Clamp ◽  
Somatosensory Cortex ◽  
Brain Slices ◽  
Patch Clamp Recording ◽  
Cortical Cells ◽  
Ventrobasal Thalamus ◽  
Layer 4 ◽  
Fast Spiking ◽  
Minimal Stimulation ◽  
Stimulation Of

Thalamocortical (TC) cells in the ventrobasal thalamus make direct excitatory connections with regular-spiking (RS) cells in layer 4 of the somatosensory cortex, but also make disynaptic feedforward inhibitory connections with the RS cells by layer 4 fast-spiking (FS) cells. In this study, we investigated connection rules of the feedforward inhibitory circuit from multiple TC cells to multiple RS cells, at the level of synaptic potentials. Using thalamocortical brain slices of young mice (postnatal days 12–16), we made simultaneous patch-clamp recordings from three adjacent cortical cells (two RS cells and one FS cell), combined with minimal stimulation of presumed single TC fibers. We found that nearly all (97%) of TC fibers, which generated excitatory inputs onto RS cells, also generated divergent excitatory inputs onto adjacent FS cells. Some 44% of TC fibers generated divergent excitatory inputs onto adjacent pairs of RS cells. We then combined the triple patch-clamp recording with multisite (two to three) minimal stimulation of single TC fibers and found that 86% of FS cells received convergent inputs from all of the stimulated TC fibers. We also found that 68% of FS cells generated divergent inhibitory inputs onto adjacent pairs of RS cells. The results indicate that spikes in TC cells, which excite RS cells, also excite adjacent FS cells with high fidelity. The results also indicate that FS cells receive convergent excitatory inputs from multiple TC cells and then send divergent inhibitory outputs to multiple RS cells.

scholarly journals Optogenetic stimulation of long-range inputs and functional characterization of connectivity in patch-clamp recordings in mouse brain slices

2018 ◽  
Author(s):  
Louis Richevaux ◽  
Louise Schenberg ◽  
Mathieu Beraneck ◽  
Desdemona Fricker
Keyword(s):  
Patch Clamp ◽  
Long Range ◽  
Brain Slices ◽  
Cell Type ◽  
Patch Clamp Recording ◽  
Axon Terminals ◽  
Optogenetic Stimulation ◽  
Cell Type Specific ◽  
The Brain ◽  
Stimulation Of

Knowledge of cell type specific synaptic connectivity is a crucial prerequisite for understanding brain wide neuronal circuits. The functional investigation of long-range connections requires targeted recordings of single neurons combined with the specific stimulation of identified distant inputs. This is often difficult to achieve with conventional, electrical stimulation techniques, because axons from converging upstream brain areas may intermingle in the target region. The stereotaxic targeting of a specific brain region for virus-mediated expression of light sensitive ion channels allows to selectively stimulate axons coming from that region with light. Intracerebral stereotaxic injections can be used in well-delimited structures, such as the anterodorsal thalamic nuclei, and also in other subcortical or cortical areas throughout the brain. Here we describe a set of techniques for precise stereotaxic injection of viral vectors expressing channelrhodopsin in the anterodorsal thalamus, followed by photostimulation of their axon terminals in hippocampal slices. In combination with whole-cell patch clamp recording from a postsynaptically connected presubicular neuron, photostimulation of thalamic axons allows the detection of functional synaptic connections, their pharmacological characterization, and the evaluation of their strength in the brain slice preparation. We demonstrate that axons originating in the anterodorsal thalamus ramify densely in presubicular layers 1 and 3. The photostimulation of Chronos expressing thalamic axon terminals in presubiculum initiates short latency postsynaptic responses in a presubicular layer3 neuron, indicating a monosynaptic connection. In addition, biocytin filling of the recorded neuron and posthoc revelation confirms the layer localization and pyramidal morphology of the postsynaptic neuron. Taken together, the optogenetic stimulation of long-range inputs in ex vivo brain slices is a useful method to determine the cell-type specific functional connectivity from distant brain regions.

Patch Clamp Recording in Brain Slices

2015 ◽  
Author(s):  
Luke Campagnola ◽  
Paul Manis
Keyword(s):  
Patch Clamp ◽  
Brain Slices ◽  
Neuronal Membrane ◽  
Electrical Noise ◽  
Patch Clamp Recording ◽  
Experimental Conditions ◽  
Neural Substrate ◽  
Hardware Configuration ◽  
Technical Issues

Patch clamp recording in brain slices allows unparalleled access to neuronal membrane signals in a system that approximates the in-vivo neural substrate while affording greater control of experimental conditions. In this chapter we discuss the theory, methodology, and practical considerations of such experiments including the initial setup, techniques for preparing and handling viable brain slices, and patching and recording signals. A number of practical and technical issues faced by electrophysiologists are also considered, including maintaining slice viability, visualizing and identifying healthy cells, acquiring reliable patch seals, amplifier compensation features, hardware configuration, sources of electrical noise and table vibration, as well as basic data analysis issues and some troubleshooting tips.

scholarly journals Combined single cell RT-PCR analysis with patch clamp recording in acute brain slices

2000 ◽  
Vol 82 ◽  
pp. 12
Author(s):  
Keisuke Tsuzuki ◽  
Bertrand Lambolez ◽  
Etienne Audinat ◽  
James T. Porter ◽  
Bruno Cauli ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Single Cell ◽  
Brain Slices ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Patch Clamp Recording

scholarly journals Local circuit input to the medullary reticular formation from the rostral nucleus of the solitary tract

2008 ◽  
Vol 295 (5) ◽  
pp. R1391-R1408 ◽  
Author(s):  
J. Nasse ◽  
D. Terman ◽  
S. Venugopal ◽  
G. Hermann ◽  
R. Rogers ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Reticular Formation ◽  
Calcium Imaging ◽  
Neonatal Rat ◽  
Solitary Tract ◽  
Patch Clamp Recording ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Stimulation Of

The intermediate reticular formation (IRt) subjacent to the rostral (gustatory) nucleus of the solitary tract (rNST) receives projections from the rNST and appears essential to the expression of taste-elicited ingestion and rejection responses. We used whole cell patch-clamp recording and calcium imaging to characterize responses from an identified population of prehypoglossal neurons in the IRt to electrical stimulation of the rNST in a neonatal rat pup slice preparation. The calcium imaging studies indicated that IRt neurons could be activated by rNST stimulation and that many neurons were under tonic inhibition. Whole cell patch-clamp recording revealed mono- and polysynaptic projections from the rNST to identified prehypoglossal neurons. The projection was primarily excitatory and glutamatergic; however, there were some inhibitory GABAergic projections, and many neurons received excitatory and inhibitory inputs. There was also evidence of disinhibition. Overall, bath application of GABAA antagonists increased the amplitude of excitatory currents, and, in several neurons, stimulation of the rNST systematically decreased inhibitory currents. We have hypothesized that the transition from licks to gapes by natural stimuli, such as quinine monohydrochloride, could occur via such disinhibition. We present an updated dynamic model that summarizes the complex synaptic interface between the rNST and the IRt and demonstrates how inhibition could contribute to the transition from ingestion to rejection.

scholarly journals Patch-clamp recordings of rat neurons from acute brain slices of the somatosensory cortex during magnetic stimulation

2014 ◽  
Vol 8 ◽  
Author(s):  
Tamar Pashut ◽  
Dafna Magidov ◽  
Hana Ben-Porat ◽  
Shuki Wolfus ◽  
Alex Friedman ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Somatosensory Cortex ◽  
Magnetic Stimulation ◽  
Brain Slices

scholarly journals Cross-Whisker Adaptation of Neurons in Layer 2/3 of the Rat Barrel Cortex

2021 ◽  
Vol 15 ◽  
Author(s):  
Yonatan Katz ◽  
Ilan Lampl
Keyword(s):  
Cortical Neurons ◽  
Barrel Cortex ◽  
Receptive Fields ◽  
Repetitive Stimulation ◽  
Complex Environments ◽  
Cortical Cells ◽  
High Responsiveness ◽  
Layer 2 ◽  
Layer 4 ◽  
Stimulation Of

Neurons in the barrel cortex respond preferentially to stimulation of one principal whisker and weakly to several adjacent whiskers. Such integration exists already in layer 4, the pivotal recipient layer of thalamic inputs. Previous studies show that cortical neurons gradually adapt to repeated whisker stimulations and that layer 4 neurons exhibit whisker specific adaptation and no apparent interactions with other whiskers. This study aimed to study the specificity of adaptation of layer 2/3 cortical cells. Towards this aim, we compared the synaptic response of neurons to either repetitive stimulation of one of two responsive whiskers or when repetitive stimulation of the two whiskers was interleaved. We found that in most layer 2/3 cells adaptation is whisker-specific. These findings indicate that despite the multi-whisker receptive fields in the cortex, the adaptation process for each whisker-pathway is mostly independent of other whiskers. A mechanism allowing high responsiveness in complex environments.

scholarly journals Automatic deep learning driven label-free image guided patch clamp system for human and rodent in vitro slice physiology

2020 ◽  
Author(s):  
Krisztian Koos ◽  
Gáspár Oláh ◽  
Tamas Balassa ◽  
Norbert Mihut ◽  
Márton Rózsa ◽  
...  
Keyword(s):  
Image Analysis ◽  
Deep Learning ◽  
Patch Clamp ◽  
Brain Research ◽  
Brain Slices ◽  
Cell Detection ◽  
Label Free ◽  
Patch Clamp Recording ◽  
Tissue Slices

ABSTRACTPatch clamp recording of neurons is a labor-intensive and time-consuming procedure. We have developed a tool that fully automatically performs electrophysiological recordings in label-free tissue slices. The automation covers the detection of cells in label-free images, calibration of the micropipette movement, approach to the cell with the pipette, formation of the whole-cell configuration, and recording. The cell detection is based on deep learning. The model was trained on a new image database of neurons in unlabeled brain tissue slices. The pipette tip detection and approaching phase use image analysis techniques for precise movements. High-quality measurements were performed on hundreds of human and rodent neurons. We also demonstrate that further molecular and anatomical analysis can be performed on the recorded cells. The software has a diary module that automatically logs patch clamp events. Our tool can multiply the number of daily measurements to help brain research.ONE SENTENCE SUMMARYNovel deep learning and image analysis algorithms for automated patch clamp systems to reliably measure neurons in human and rodent brain slices.

Patch Clamp Recording in Vivo

2015 ◽  
Author(s):  
David Ferster
Keyword(s):  
Patch Clamp ◽  
Brain Slices ◽  
Mammalian Brain ◽  
Intracellular Recordings ◽  
Intracellular Membrane ◽  
Patch Clamp Recording ◽  
Intact Organism ◽  
Technological Developments ◽  
History Of

Patch clamp recording in vivo allows an investigator to study intracellular membrane potentials in an intact organism (as opposed to cells in culture or acute brain slices). This technique is a reliable method of obtaining high-quality intracellular recordings from neurons, regardless of their size, in several parts of the mammalian brain. This chapter will describe the principles and practice of performing patch clamp experiments in vivo, beginning with a brief history of the technological developments that have made this technique possible.

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