A critical role for NMDA receptors in parvalbumin interneurons for gamma rhythm induction and behavior methods

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    "name": "A critical role for NMDA receptors in parvalbumin interneurons for gamma rhythm induction and behavior methods",
    "description": "Protocol for A critical role for NMDA receptors in parvalbumin interneurons for gamma rhythm induction and behavior methods",
    "step": [
      {
        "@type": "HowToStep",
        "position": 1,
        "name": "Slice electrophysiology",
        "text": "AAV DIO channelrhodopsin-2 (ChR2)-mCherry was injected into hippocampus of 5- to 7-week-old PV-Cre and PV-Cre/NR1f/f mice. At 7-10 days after viral transduction, transverse hippocampal slices (400 µ m thickness) were prepared as described., The tungsten bipolar electrode (FHC, Bowdoin, ME, USA) was placed in the stratum radiatum or oriens and the Schaffer collateral/commissural fibers were stimulated at 0.1 Hz. Picrotoxin (0.15 m M, Sigma, St Louis, MO, USA) was dissolved in artificial cerebrospinal fluid (aCSF) to block GABA A receptor-mediated synaptic transmission for whole-cell patch clamp recordings. α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor-mediated excitatory postsynaptic current (EPSCs) were recorded at -70 mV and NMDAR-mediated EPSCs were recorded at +40 mV with the same stimulus strength in the presen..."
      },
      {
        "@type": "HowToStep",
        "position": 2,
        "name": "Anesthetized electrophysiology",
        "text": "AAV DIO ChR2-mCherry was injected into barrel cortex of adult (8-12 weeks old) PV-Cre or PV-Cre/NR1f/f mice as described earlier. Electrophysiological in vivo recordings were performed 1-3 weeks after viral injections. Extracellular single-unit and local field potential (LFP) recordings were made with tetrodes or stereotrodes. Stimulus control and data acquisition was performed using software custom written in LabView (National Instruments, Austin, TX, USA) and Matlab (The Mathworks, Natick, MA, USA) by Ulf Knoblich."
      },
      {
        "@type": "HowToStep",
        "position": 3,
        "name": "Awake electrophysiology",
        "text": "Eight NR1f/f control and eight PV-Cre/NR1f/f mice (8-10 weeks) were used for awake electrophysiology. Teflon-coated tungsten electrodes (impedance of 100 kΩ) were implanted: two electrodes were placed bilaterally in primary somatosensory (barrel) cortices 1.5 mm posterior to bregma and 3.5 mm from the midline. In each hemisphere, a signal electrode was implanted 0.5 mm below the cortical surface and a reference electrode was implanted 1.75 mm below the cortical surface. A stainless steel screw over right posterior parietal cortex served as ground. Recording sessions took place in an empty box to which animals had not been previously exposed. After a 5-10-min habituation period, 20 min of baseline data were recorded for each animal. After the baseline period, four NR1f/f control and four PV-Cre/NR1f/f mice were injected intraperitone..."
      },
      {
        "@type": "HowToStep",
        "position": 4,
        "name": "Characterization of PV-Cre/NR1f/f mice",
        "text": "PV-Cre/NR1f/f mice were viable, developed normally and did not exhibit growth abnormalities or other gross anatomical changes (data not shown). Cre-dependent recombination from the PV locus was specific to PV-expressing cells and followed the postnatal onset of PV expression ( ). PV-Cre-driven recombination in somatosensory cortex and hippocampus was detected at postnatal day 13 (P13) with increased recombination at 29 days (P29) and almost complete recombination at 8 weeks (, ). Whole-cell recordings in hippocampal slices in vitro confirmed the functional loss of NMDAR currents in PV cells in PV-Cre/NR1f/f mice (five cells in four PV-Cre/NR1f/f mice, seven cells in five control mice, P =0.03, unpaired t -test; ). We found the cortical architecture (layers and barrels in somatosensory cortex), migration and differentiation of PV interneurons to be normal in PV-Cre/NR1f/f mice (; P =..."
      },
      {
        "@type": "HowToStep",
        "position": 5,
        "name": "Spontaneous and induced gamma oscillations are altered in PV-Cre/NR1f/f mice",
        "text": "To explore the role of NMDAR in PV interneurons on evoked gamma activity in the awake state, we challenged PV-Cre/NR1f/f mice with an acute administration of the non-competitive NMDAR antagonist MK-801 (0.5 mg kg -1; intraperitoneal; ) or saline (data not shown). In agreement with previous findings, control mice acutely displayed a significant increase in LFP in the 30-50 Hz gamma range (post1, 5-15 min after MK-801 administration; P <0.05, Wilcoxon signed-rank test; ). PV-Cre/NR1f/f mice, in contrast, displayed a significant reduction in gamma-band activity after NMDAR antagonist treatment ( ). Control mice and PV-Cre/NR1f/f mice displayed a significant induction in the 6-10 Hz frequency band relative power at later time points (post2, 25-35 min after MK-801 administration; ). In the 12-24 Hz band, the rel..."
      },
      {
        "@type": "HowToStep",
        "position": 6,
        "name": "Behavioral effects of NMDAR deletion in PV cells",
        "text": "We characterized the PV-Cre/NR1f/f mice in a series of paradigms to assess general locomotor and exploratory behaviors as well as cognitive tasks evaluating learning and memory. The PV-Cre/NR1f/f mice and their littermate controls (NR1f/f) were introduced into an automated novel open field environment at an age of 7 weeks ( n =11 per genotype). We did not observe any consistent behavioral differences between PV-Cre/NR1f/f and control mice over the entire 60 min trial or in any 5 min block after analysis of 18 different parameters, including total distance ( ), locomotion, stereotypy and center/margin time (data not shown)."
      },
      {
        "@type": "HowToStep",
        "position": 7,
        "name": "Behavioral effects of NMDAR deletion in PV cells",
        "text": "We questioned whether PV-Cre/NR1f/f mice at later stages develop behavioral abnormalities in the open field and therefore assessed mice of 11-12 weeks of age ( n =9 per genotype). We found a nonsignificant trend toward a decrease in total distance traveled over the 60-min period for PV-Cre/NR1f/f mice compared with control mice ( P >0.05; Mann-Whitney test; ). Analysis of time spent in the center ( ) or margin of the open field, vertical movements (data not shown) or stereotypy counts ( ) did not reveal differences between the genotypes over the 60-min period."
      },
      {
        "@type": "HowToStep",
        "position": 8,
        "name": "Behavioral effects of NMDAR deletion in PV cells",
        "text": "Our electrophysiological findings indicate that the PV-Cre/NR1f/f mice have a selective deficit in gamma emergence following optogenetic drive of FS-PV interneurons or pharmacological NMDAR inhibition. To investigate potential behavioral differences in response to acute NMDAR inhibition, we exposed naïve PV-Cre/NR1f/f and control mice ( n =11 per genotype) to the open field for 30 min and then challenged them with administration of the NMDAR antagonist MK-801 at a low dose (0.3 mg kg -1 ) followed by continued exposure to the open field for 60 min. Control mice responded over time to MK-801 administration with increased horizontal activity and induction of stereotypies as previously reported ( ). In contrast, PV-Cre/NR1f/f mice showed a markedly reduced sensitivity to MK801 ( ), consistent with our electrophysiological findings that PV interneu..."
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    "isBasedOn": {
      "@type": "ScholarlyArticle",
      "headline": "A critical role for NMDA receptors in parvalbumin interneurons for gamma rhythm induction and behavior",
      "datePublished": "2012",
      "author": [
        {
          "@type": "Person",
          "name": "M Carlén"
        },
        {
          "@type": "Person",
          "name": "K Meletis"
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        {
          "@type": "Person",
          "name": "J H Siegle"
        },
        {
          "@type": "Person",
          "name": "J A Cardin"
        },
        {
          "@type": "Person",
          "name": "K Futai"
        },
        {
          "@type": "Person",
          "name": "D Vierling-Claassen"
        },
        {
          "@type": "Person",
          "name": "C Rühlmann"
        },
        {
          "@type": "Person",
          "name": "S R Jones"
        },
        {
          "@type": "Person",
          "name": "K Deisseroth"
        },
        {
          "@type": "Person",
          "name": "M Sheng"
        },
        {
          "@type": "Person",
          "name": "C I Moore"
        },
        {
          "@type": "Person",
          "name": "L-H Tsai"
        }
      ],
      "identifier": "10.1038/mp.2011.31"
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