Perineuronal nets stabilize the grid cell network methods
Aim. Evidence-backed execution summary for Perineuronal nets stabilize the grid cell network methods from Perineuronal nets stabilize the grid cell network.
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rat
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Surgical procedures
All surgical procedures were performed in an aseptic environment. Rats were anesthetized with isoflurane mixed with air (5% induction, 1.5-2% for maintenance) and immobilized in a stereotaxic frame (World Precision Instruments Ltd, Hertfordshire, UK). They were given subcutaneous injections of buprenorphine (0...
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- All surgical procedures were performed in an aseptic environment. Rats were anesthetized with isoflurane mixed with air (5% induction, 1.5-2% for maintenance) and immobilized in a stereotaxic frame (World Precision Instruments Ltd, Hertfordshire, UK). They were given subcutaneous injections of buprenorphine (0...
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Grid cell representations in familiar environments
In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores (Fig. c). Pharmacogenetically reducing activity in PV + neurons lead to increased out-of-field activity in grid cells. To test if similar effects could be seen after removal of PNNs, we identified firing fields for each grid cell (Supplementary Fig. ) and calculated mean firing rate inside and outside of fields. Grid cells from chABC-treated rats showed a weak but consistent tendency of reduced firing rates inside grid fields and increased firing rates outside grid fields (Supplementary Table ). The combination of these effects lead to a prominent decrease in spatial specificity of grid cells in animals with disrupted PNNs (Fig. d). To account for effects of an arbitrary definition of g...
Methods
This study used recording data from 23 male Long-Evans rats (3-8 months old, 350-550 g at surgery). After surgeries the animals were housed individually in transparent Plexiglas cages (45 × 30 × 35 cm) in a temperature- and humidity-controlled vivarium. All rats were maintained on a 12-h light/12-h dark schedule. Testing occurred in the dark phase. The rats were kept at 85-90% of free-feeding body weight and food deprived 18-24 h before each training and recording trial. Water was available ad libitum. Experiments were performed in accordance with the Norwegian Animal Welfare Act and the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes.
Surgical procedures
All surgical procedures were performed in an aseptic environment. Rats were anesthetized with isoflurane mixed with air (5% induction, 1.5-2% for maintenance) and immobilized in a stereotaxic frame (World Precision Instruments Ltd, Hertfordshire, UK). They were given subcutaneous injections of buprenorphine (0.04 mg/kg) and local subcutaneous injections of bupivacaine/adrenaline (Marcain adrenaline, 13.2 mg/kg) in the scalp before surgery began. The scalp was shaved and cleaned with ethanol and chlorhexidine. Heart rate and core temperature were continuously monitored throughout the operation through a MouseStat system (Kent Scientific, CT, USA), the latter in a feedback mechanism to a heating pad. In addition, the hind paw withdrawal reflex was used to assess the depth of anesthesia.
Surgical procedures
Tetrodes were implanted above MEC at AP 0.4 ± 0.1 mm in front of the transverse sinus and ML 4.5 ± 0.1 mm relative to the midline. Tetrodes implanted above hippocampus were placed at AP Bregma -3.8 ± 0.2 mm and ML 3.1 ± 0.1 mm. The depth of implantation was 1800 µm measured from the surface of dura. Jeweler's screws fixed to the skull served as ground electrodes. The microdrives were secured to the skull using jeweler's screws and dental cement. All animals were given a subcutaneous injection of carprofen (5 mg/kg) at the end of the surgery, and the edge of the wound was cleaned and local anesthetic ointment Lidocain was applied. This was repeated for 3 days after surgery.
Extracellular recordings
First, a recording session of 20 min was conducted in the familiar environment followed by 3 × 20 min recording sessions in a novel environment. Lastly, the animals were put back in the familiar environment for an additional 20 min session. The animals rested for 5-10 min in the home cage between sessions. This experimental procedure was repeated for three consecutive days, with the exception that the novel environment recording on days 2 and 3 consisted of only one 20 min long recording session.
Tissue processing and immunohistochemistry
At the end of the experiments, animals were deeply anesthetized by an intraperitoneal injection of pentobarbital sodium (50 mg/kg) and intracardially perfused with 0.9% NaCl followed by 4% paraformaldehyde (PFA) in 1 × PBS. The brains were dissected out and post-fixed in 4% PFA overnight. They were cryoprotected in 30% sucrose in 1 × PBS for 3 days, and 40 µm sagittal (MEC) or coronal (hippocampus) sections were cut with a cryostat. Staining procedures were performed on free-floating sections under constant agitation unless mentioned otherwise. The lectin WFA was used to visualize PNNs.
Coefficient of variation
To reduce bias in ISI that arises from animals traveling between receptive fields, we calculated only in-field CV-values by extracting one CV value for each pass through the fields. This was compared with two other methods. In the first, a CV value was calculated for the entire session. The second followed the protocol of ref., where sections of the recording where the speed of the rat was less than 8 cm/s were selected. A CV value was then calculated for the sections with a longer duration than 0.2 s and at least two spikes. See Supplementary Fig. for comparison of the three methods.
Measurement outputs
What raw and processed outputs should exist?
In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
As animals move through the environment, grid cells are thought to be updated by path integration through cells that monitor the animal's instantaneous speed. The impairm...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
The PSD were calculated on z -scored LFP signal using the Welch method given by mlab.psd from the Matplotlib package.
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
- Structured table with cleaned measurements ready for comparison
- Reported as
- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
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Acquisition
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inferred from protocolPreprocessing / cleaning
In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores (Fig. c).
from paperScoring or quantification
Quantify the primary readouts for this experiment: In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores...; As animals move through the environment, grid cells are thought to be updated by path integration through cells that monitor the animal's instantaneous speed. The impairm...; The PSD were calculated on z -scored LFP signal using the Welch method given by mlab.psd from the Matplotlib package..
from paperStatistical comparison
In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores...; Grid cell pairs were selected from the same recording and hemisphere. Temporal pairwise cross-correlations were calculated by first calculate the instantaneous firing rate by co...
from paperReporting output
Report representative outputs alongside summary comparisons for In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores..., As animals move through the environment, grid cells are thought to be updated by path integration through cells that monitor the animal's instantaneous speed. The impairm..., The PSD were calculated on z -scored LFP signal using the Welch method given by mlab.psd from the Matplotlib package..
inferred from protocolStructured statistical methods
In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores...; Grid cell pairs were selected from the same recording and hemisphere. Temporal pairwise cross-correlations were calculated by first calculate the instantaneous firing rate by co...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (7)
In animals treated with chABC we observed more dispersed spiking within and at the edges of grid fields (Fig. a, b), but this did not lead to reduction in gridness scores (Fig. c). Pharmacogenetically reducing activity in PV + neurons lead to increased out-of-field activity in grid cells. To test if similar effects could be seen after removal of PNNs, we identified firing fields for each grid cell (Supplementary Fig. ) and calculated mean firing rate inside and outside of fields. Grid cells from chABC-treated rats showed a weak but consistent tendency of reduced firing rates inside grid fields and increased firing rates outside grid fields (Supplementary Table ). The combination of these effects lead to a prominent decrease in spatial specificity of grid cells in animals with disrupted PNNs (Fig. d). To account for effects of an arbitrary definition of grid fields, we also calculated the spatial information content of each grid cell, a measure that is independent of any predefined firing fields. The spatial information was significantly reduced in grid cells from animals treated with chABC (Fig. e). In addition, we found changes in the tempo...
This study used recording data from 23 male Long-Evans rats (3-8 months old, 350-550 g at surgery). After surgeries the animals were housed individually in transparent Plexiglas cages (45 × 30 × 35 cm) in a temperature- and humidity-controlled vivarium. All rats were maintained on a 12-h light/12-h dark schedule. Testing occurred in the dark phase. The rats were kept at 85-90% of free-feeding body weight and food deprived 18-24 h before each training and recording trial. Water was available ad libitum. Experiments were performed in accordance with the Norwegian Animal Welfare Act and the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes.
All surgical procedures were performed in an aseptic environment. Rats were anesthetized with isoflurane mixed with air (5% induction, 1.5-2% for maintenance) and immobilized in a stereotaxic frame (World Precision Instruments Ltd, Hertfordshire, UK). They were given subcutaneous injections of buprenorphine (0.04 mg/kg) and local subcutaneous injections of bupivacaine/adrenaline (Marcain adrenaline, 13.2 mg/kg) in the scalp before surgery began. The scalp was shaved and cleaned with ethanol and chlorhexidine. Heart rate and core temperature were continuously monitored throughout the operation through a MouseStat system (Kent Scientific, CT, USA), the latter in a feedback mechanism to a heating pad. In addition, the hind paw withdrawal reflex was used to assess the depth of anesthesia.
Tetrodes were implanted above MEC at AP 0.4 ± 0.1 mm in front of the transverse sinus and ML 4.5 ± 0.1 mm relative to the midline. Tetrodes implanted above hippocampus were placed at AP Bregma -3.8 ± 0.2 mm and ML 3.1 ± 0.1 mm. The depth of implantation was 1800 µm measured from the surface of dura. Jeweler's screws fixed to the skull served as ground electrodes. The microdrives were secured to the skull using jeweler's screws and dental cement. All animals were given a subcutaneous injection of carprofen (5 mg/kg) at the end of the surgery, and the edge of the wound was cleaned and local anesthetic ointment Lidocain was applied. This was repeated for 3 days after surgery.
First, a recording session of 20 min was conducted in the familiar environment followed by 3 × 20 min recording sessions in a novel environment. Lastly, the animals were put back in the familiar environment for an additional 20 min session. The animals rested for 5-10 min in the home cage between sessions. This experimental procedure was repeated for three consecutive days, with the exception that the novel environment recording on days 2 and 3 consisted of only one 20 min long recording session.
At the end of the experiments, animals were deeply anesthetized by an intraperitoneal injection of pentobarbital sodium (50 mg/kg) and intracardially perfused with 0.9% NaCl followed by 4% paraformaldehyde (PFA) in 1 × PBS. The brains were dissected out and post-fixed in 4% PFA overnight. They were cryoprotected in 30% sucrose in 1 × PBS for 3 days, and 40 µm sagittal (MEC) or coronal (hippocampus) sections were cut with a cryostat. Staining procedures were performed on free-floating sections under constant agitation unless mentioned otherwise. The lectin WFA was used to visualize PNNs.
To reduce bias in ISI that arises from animals traveling between receptive fields, we calculated only in-field CV-values by extracting one CV value for each pass through the fields. This was compared with two other methods. In the first, a CV value was calculated for the entire session. The second followed the protocol of ref., where sections of the recording where the speed of the rat was less than 8 cm/s were selected. A CV value was then calculated for the sections with a longer duration than 0.2 s and at least two spikes. See Supplementary Fig. for comparison of the three methods.
Machine-readable layer
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