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University of Nottingham Medical School Neuronal Networks Laboratory Using Plexon at Nottingham: Sensory Gating, Anxiety, Seizures, and Pain Dr Rob Mason.

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Presentation on theme: "University of Nottingham Medical School Neuronal Networks Laboratory Using Plexon at Nottingham: Sensory Gating, Anxiety, Seizures, and Pain Dr Rob Mason."— Presentation transcript:

1 University of Nottingham Medical School Neuronal Networks Laboratory Using Plexon at Nottingham: Sensory Gating, Anxiety, Seizures, and Pain Dr Rob Mason School of Biomedical Sciences University of Nottingham Medical School Neuronal Networks Laboratory

2 Talk Format Brief overview of what we have been studying using Plexon system(s) at Nottingham Indicate on route some hardware developments & signal processing / analysis approaches we use schizophrenia – sensory gating epilepsy anxiety pain Neuronal Networks Laboratory

3 Multiple Electrode Recordings in vivo simultaneous multichannel spike & field potential recording 64-channel MAP system 32-channel MAP system – with CinePlex for behavioural studies 64-channel Recorder system 16-channel Recorder system used MAP & Recorder for in vitro MEA studies with brain slices & neuronal cultures

4 LAB TEAM epilepsy Ben Coomber [Dr Mike ODonoghue] Clare Roe schizophrenia Dr Jill Suckling [Prof CA Marsden] Dr Dissanayake anxiety Dr Carl Stevenson [Prof CA Marsden] pain Dr Steve Elmes [Dr V Chapman] rodent USVsBeth Tunstall[Dr S Beckett] data analysisMargarita Zachariou[Dr S Coombes / Dr M Owen] [Mathematics Dept] Dr David Halliday (University of York) Prof D Auer (fMRI / phMRI) Neuronal Networks Laboratory

5 LAB EXPERIMENTAL DIRECTIONS mPFCx interconnectivity & functional context – in vivo studies maternal separation - depression circadian - affective states sensory gating - schizophrenia - epilepsy USVs - affective states medial Pre-Frontal Cortex Amygdala nucleus AccumbensHippocampus SCN Contralateral mPFC VTA - schizophrenia - drugs of abuse PVt

6 Dual site recordings VTA - mPFC hippocampus - mPFC amygdala - mPFC mPFC - mPFC Systemic pharmacological manipulation Local pharmacological manipulation injec-trode integrated microiontophoresis with recording array Electrical stimulation Independent electrode array microdrive Anaesthetised & awake-behaving preparations Neuronal Networks Laboratory

7 University of Nottingham Medical School Neuronal Networks Laboratory Project #1: Schizophrenia auditory-evoked sensory gating effects of PCP / ketamine - model effects of social isolation - model Neuronal Networks Laboratory

8 Schizophrenia - VTA/mPFCx dual-site recordings medial prefrontal cortex - mPFCventral tegmental area - VTA Prussian Blue reaction - locate recording site(s) - histology PFC VTA

9 Dual site recording: pre-frontal cortical / VTA recording multiple single-units & LFPs 1 mV 5 s VTA cells PFC cells VTA LFP PFC LFP Neurone # Matlab scripts for off-line characterisation unit spike waveforms & LFPs

10 Explore connectivity between PFC and VTA - e.g. local application of drug ( e.g. glutamate) to VTA to drive mPFC Local application of drugs – injec-trode 10s Extra-VTA PFC Extra-VTA LFP PFC LFP 50 nl Glutamate (100 mM) Electrode headstage Tubing attached to Hamilton syringe Electrode tips Cannula tip

11 Sensory gating in hippocampus: A model for schizophrenia ? Sensory gating: mechanism(s) by which irrelevant sensory information is filtered – enables efficient information processing Auditory Conditioning-Test paradigm: measures reduction in auditory-evoked response produced by Test stimulus following a Conditioning stimulus Stimuli: 3kHz sine-wave / 10ms duration / presented 500ms apart / 80-90dB human P50 wave = rat N40 component Gating absent in- schizophrenic patients (& near family members) - normal human volunteers given PCP / amphetamine - rats given PCP / amphetamine Neuronal Networks Laboratory

12 1s LFP2 averaged 128 trials Units LFPs Hippocampal CA3 - auditory-evoked unit & LFP activity Trial # s Cs Ts mV N40 Averaged LFP T/C ratio = 50% event tone

13 dentate gyrus CA3 region Single-unit PSTHs rasters histograms - gating rats Unit 1 Unit 2 Unit 3 Unit 4 Spike raster PSTH LFP LFP trial raster

14 Hippocampal CA3 auditory-evoked unit & LFP activity Effects of PCP (1mg/kg i.p.) attenuates/abolishes sensory gating Basal [128 trials] T/C ratio = 32% 45mins after PCP T/C ratio = 66% 1 Trail # 128

15 University of Nottingham Medical School Neuronal Networks Laboratory Project #2: Epilepsy kainate-induced epileptiform activity – TLE functional network interactions - hippocampus mPFC electrical stimulation – perforant pathway role of endocannibnoid system – CB 1 R pharmacology Neuronal Networks Laboratory

16 hippocampus mPFC PFC Hipp PFC Hipp L Hipp M Basal firingKA-induced firing (10mg kg -1, i.p.) 2s Kainate-induced epileptiform activity – dual recording hippocampus & mPFC

17 mPFC Hipp mPFC HIPP Hippocampal-mPFC – functional connectivity Burst analysis – Synchrony index - Cross-correlation – coherence - PDC Ref unit mPFCHippocampus unit 3H unit 1H unit 2H

18 PDC Analysis – Unit populations Identify direction of activity between hippocampus and mPFC - PDC applied: technique with potential to reveal the neuronal ensemble drive direction. Note: the magnitude of the classical coherence gives no information about directional connectivity, but its phase may do so. KA

19 PDC Analysis – LFPs Representative PDC plot between LFPs recorded from 3 electrodes in both mPFC (AD01-08) hippocampus (AD10 to AD16 = medial to lateral electrodes). TIME FRAME # Hippocampus mPFC Hippocampus Red boxes highlight PDC between brain regions – suggesting the predominant flow is from mPFC to hippocampus Green boxes highlight that the predominant flow within hippocampus is from lateral to medial.

20 LFPanalyser suite 2006 – custom Matlab script Basal (pre-KA challenge) - hippocampus

21 45min post-KA - hippocampus LFPanalyser suite 2006

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23 % Power DistributionBasal 45min Post-KA LFPanalyser suite 2006

24 University of Nottingham Medical School Neuronal Networks Laboratory Project #3: Nociception & pain management dual spinal cord / supraspinal recording Role of endocannabinoid system in normal physiology and pain (e.g. neuropathic) states Neuronal Networks Laboratory

25 innocuous (7g) stimulation noxious (65g) stimulation Mechanically-evoked response in somatosensory thalamus (VPM)

26 University of Nottingham Medical School Neuronal Networks Laboratory Project #4: Affective state Cortico-limibic network interactions anxiety effects of maternal separation pharmacologically-induced (e.g. FG-7142) anxiety behavioural sequalae rodent ultrasound vocalisations Neuronal Networks Laboratory

27 Role of nuc accumbens in USV-mediated Behaviours nucleus accumbens / basolateral amygdala - mPFCx functional connectivity nucleus accumbens – mPFC Rat ultrasound vocalisations (USVs) & affective state Neuronal Networks Laboratory 50kH z 22kHz Time (s) Frequency (kHz) ~22 kHz calls - aversive stimuli (call Type G) ~50 kHz calls - rewarding stimuli (Types A-F)

28 Combined unit / LFP with USV / behavioural recording USV call start time unit activity - nuc accumbens USV Recorder input Behavioural data Local Field Potential (LFP) Spectrogram of specific call s [AviSoft]

29 Multiple Electrode Recordings in vivo simultaneous multichannel spike / field potential / behavioural recording Video camera CinePlex system

30 Behavioural Electrophysiology II Circular arena recording using CinePlex Movie - rat HopScotch: 8-microwire array in nuc. accumbens - 6 weeks post implant Neuronal Networks Laboratory Recorded video Neural data

31 University of Nottingham Medical School Neuronal Networks Laboratory Lab web site Neuronal Networks Laboratory Thats all folks

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