ds3  (Digitimer North America LLC)

Journal: Science Advances

Article Title: Beyond seizure control: Identifying deficits in cognitive networks in absence epilepsy

doi: 10.1126/sciadv.aed3642

Figure Lengend Snippet: ( A ) Left: Recording configuration for LFPs. A 16-channel silicon probe was positioned across cortical layers of the mPFC in acute coronal slices from Scn8a +/− and wild-type littermates expressing ChR2 in the Re. Right: Epifluorescence micrograph showing the eYFP/ChR2 fibers (green) from the Re projecting to the mPFC. ( B ) Example CSD profiles evoked by optogenetic Re activation. ( C ) Peak CSD sinks and sources in L2/3 (top) and L5 (bottom) of the mPFC. ( D ) Recording configuration for whole-cell patch clamp: L5 pyramidal neurons in mPFC were recorded in acute slices with ChR2-expressing Re afferents. ( E ) Example traces of monosynaptic EPSCs (−70 mV) and polysynaptic IPSCs (+10 mV) evoked by Re optogenetic stimulation. ( F ) E/I ratio calculated as EPSC amplitude (−70 mV) divided by IPSC amplitude (+10 mV). ( G ) Example current-clamp traces showing subthreshold EPSPs evoked by Re stimulation. ( H and I ) Quantification of EPSP rise time (H) and decay time (I).

Article Snippet: For stimulation, blue laser pulses (473 nm; Laserglow Technologies) were delivered in 1-min trains at 20 Hz (5-ms pulse duration, 50-ms pulse interval, 3.9-mW output, 3.4 mW/cm 2 ), interleaved with 5-min intervals without stimulation, and repeated throughout the recording period.

Techniques: Expressing, Activation Assay, Patch Clamp

Journal: Science Advances

Article Title: Beyond seizure control: Identifying deficits in cognitive networks in absence epilepsy

doi: 10.1126/sciadv.aed3642

Figure Lengend Snippet: ( A ) Whole-cell patch recordings from L5 pyramidal neurons in mPFC slices with a partially disconnected L1 flap. A bipolar electrode delivered electrical stimulation in the L1 flap. ( B ) Example EPSCs (−70 mV) and IPSCs (+10 mV) evoked by increasing stimulation intensities. ( C ) Quantification of EPSC and IPSC amplitudes. ( D ) Whole-cell patch recordings from L5 pyramidal neurons in acute slices from Scn8a med -VGAT-ChR2 mice. ( E ) Example IPSCs evoked by optogenetic activation of local interneurons. ( F ) Quantification of IPSC properties. ( G ) Whole-cell patch recordings from L5 pyramidal neurons in slices expressing ChR2 in Re afferents. ( H ) Example monophasic and polyphasic EPSCs evoked by optogenetic activation of Re afferents. ( I ) Proportion of L5 neurons responding to Re afferent stimulation with monophasic versus polyphasic EPSCs.

Article Snippet: For stimulation, blue laser pulses (473 nm; Laserglow Technologies) were delivered in 1-min trains at 20 Hz (5-ms pulse duration, 50-ms pulse interval, 3.9-mW output, 3.4 mW/cm 2 ), interleaved with 5-min intervals without stimulation, and repeated throughout the recording period.

Techniques: Activation Assay, Expressing

Journal: Science Advances

Article Title: Beyond seizure control: Identifying deficits in cognitive networks in absence epilepsy

doi: 10.1126/sciadv.aed3642

Figure Lengend Snippet: ( A ) Epifluorescence micrograph of an mPFC slice showing recorded L1 interneuron (purple), Re afferents expressing ChR2-eYFP (green), and DAPI-stained nuclei (blue). ( B ) Firing patterns of L1 interneurons. ( C ) Representative action potentials. ( D ) Quantification of action potential (AP) threshold and afterhyperpolarization. ( E ) EPSCs recorded in L1 interneurons. ( F ) Quantification of EPSC properties. ( G ) IPSCs recorded in L1 interneurons. ( H ) Quantification of IPSC properties. ( I ) Action potential discharge from resting membrane potential during 10-Hz train stimulation of Re afferents. ( J ) Quantification of action potential discharge probability during each stimulus within a 10-Hz train of stimuli.

Article Snippet: For stimulation, blue laser pulses (473 nm; Laserglow Technologies) were delivered in 1-min trains at 20 Hz (5-ms pulse duration, 50-ms pulse interval, 3.9-mW output, 3.4 mW/cm 2 ), interleaved with 5-min intervals without stimulation, and repeated throughout the recording period.

Techniques: Expressing, Staining, Membrane

Journal: Science Advances

Article Title: Beyond seizure control: Identifying deficits in cognitive networks in absence epilepsy

doi: 10.1126/sciadv.aed3642

Figure Lengend Snippet: ( A ) Recording configuration: EEG electrodes in the somatosensory cortex (SC) and MC, with optic fiber targeting the Re. Mice were awake and head restrained on a cylindrical treadmill. ( B ) Stimulation protocol: 10 repetitions of a 1-min, 20-Hz train followed by 5 min without stimulation (total duration, 1 hour). ( C ) EEG traces from a 1-hour recording. Blue bars mark periods of light delivery in the recording cage (left, control stimulation) or on the Re (right). ( D ) Expanded EEG traces from (C) showing a typical SWD. ( E ) Average number of seizures during 1-hour-long recordings in control and Re stimulation conditions. ( F ) Average speed on the cylindrical treadmill during 1-hour-long recordings in control and Re stimulation conditions. ( G ) Same as (E) restricted to the 10 blocks of stimulation.

Article Snippet: For stimulation, blue laser pulses (473 nm; Laserglow Technologies) were delivered in 1-min trains at 20 Hz (5-ms pulse duration, 50-ms pulse interval, 3.9-mW output, 3.4 mW/cm 2 ), interleaved with 5-min intervals without stimulation, and repeated throughout the recording period.

Techniques: Control

Journal: Science Advances

Article Title: Beyond seizure control: Identifying deficits in cognitive networks in absence epilepsy

doi: 10.1126/sciadv.aed3642

Figure Lengend Snippet: ( A ) Schematic of the swimming Y-maze task. Mice underwent a learning phase on days 1 and 2 (left) followed by a reversal phase on day 3 (right). ( B ) Learner (L) mice reached the criterion of ≥7 of 10 correct trials on day 2 and advanced to the reversal phase. Nonlearners (NL) and floaters (F) were excluded from subsequent analyses. ( C ) Escape latency during the learning phase for Scn8a +/+ (black), Scn8a +/− (red), and Scn8a +/− with Re optogenetic stimulation (day 2). ( D ) Escape latency during the reversal phase (day 3).

Article Snippet: For stimulation, blue laser pulses (473 nm; Laserglow Technologies) were delivered in 1-min trains at 20 Hz (5-ms pulse duration, 50-ms pulse interval, 3.9-mW output, 3.4 mW/cm 2 ), interleaved with 5-min intervals without stimulation, and repeated throughout the recording period.

Techniques: