To determine the effects of genotype on MK-801–induced hyperlocomotion, mice were habituated in the open field (40 cm by 40 cm by 40 cm box) for 60 min, followed by the administration of vehicle or MK-801, and locomotor activity was recorded for another 60 min. To determine the effects of MTEP and BRD0705 on MK-801–induced hyperlocomotion, mice were habituated in the open field for 30 min, followed by the intraperitoneal administration of MTEP (10 mg/kg at a dosing volume of 10 ml/kg) or BRD0705 (30 mg/kg at a dosing volume of 2 ml/kg). After an additional 30 min, MK-801 (0.3 mg/kg at a dosing volume of 10 ml/kg) was administered intraperitoneally, and locomotor activity was recorded for another 60 min. The time course of drug-induced changes in ambulation was expressed as centimeters traveled per 5 min over the 120-min session. Sessions were recorded using Plexon’s CinePlex Studio and analyzed using Plexon’s CinePlex Editor and code was written in MATLAB. MK-801–induced locomotor activity was scored and analyzed using the average of the final 5 min (minutes 115 to 120) of observation per cohort.
Cineplex studio
Manufactured by Plexon
CinePlex Studio is a comprehensive video tracking and data acquisition system from Plexon. It provides real-time video tracking and synchronization with neural and behavioral data. The system offers advanced features for 2D and 3D tracking of animal movement and behavior.
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8 protocols using cineplex studio
1
Behavioral Effects of MK-801 and Modulators
2
Auditory Fear Conditioning in Mice
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Nine days after choleratoxin-B injection to mPFC subdivisions, mice were submitted to an auditory fear conditioning paradigm. Five pairings of auditory conditioned stimulus (CS) and aversive unconditioned stimulus (US) were presented with an intertrial interval of 78-110 s. The CS consisted of 50 ms pips repeated at 0.9 Hz (total duration of 30 s) with a pip frequency of 7.5 kHz and 75 dB sound pressure level (Tucker-Davis Technologies) and was followed by a 1 s 0.65 mA AC foot shock (Coulbourn Instruments). Freezing behavior was classified as a 2 s absence of movement and quantified using Cineplex Studio and CinePlex Editor video tracking software (Plexon) and customwritten MATLAB (MathWorks) routines. Mice were perfused for immunohistochemical analysis 24 hr after fear conditioning.
3
Optogenetic Stimulation and EMG Recordings
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For electromyographic (EMG) recordings during stimulation of MLR-Rbp4 neurons, injection and fiber implantation were conducted as described above (see virus production, injections and implantations). Cable preparation and EMG implantation of the biceps and triceps muscle of the forelimb were conducted as previously reported (Miri et al., 2017 (link)). Acquisition was carried out together with optogenetic stimulation (1 s continuous light, 20mW), during locomotion on a treadmill, set to 10 cm/s to encourage continuous locomotion. The signal was amplified and bandpass filtered (A-M systems 1700, gain 100, bandpass 100-1000 Hz) and acquired using a plexon recording system (Omniplex, Plexon Inc.) at 5000 Hz. Mean subtraction was applied to correct for the DC offset. Movies were recorded from the side with a video camera (Pike, Allied Vision Inc.) acquiring at 100 fps, controlled by the Cineplex Studio software (Plexon Inc.).
4
Electrophysiological Recordings and Neural Tracking
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Electrophysiological recordings for this project were collected using a 96 channel OmniPlex D Neural Acquisition System (Plexon). Each channel was amplified and bandpass filtered for both single-unit activity (154 Hz to 8.8 kHz) and local field potentials (1.5 Hz to 400 kHz). Spike channels were referenced to a local electrode in the same region in order to remove both movement-related and any electrical noise. Action potentials of neurons were detected via threshold crossing and digitized at 40 kHz. Between recorded training sessions tetrodes were advanced based on visual inspection, in order to maximize neural unit yield, and allowed to settle overnight before conducting the next recording session. Individual neural units were isolated via manual offline clustering, employing Offline Sorter v3 (Plexon). Cineplex Studio (Plexon) was used for capturing behavioral tracking data via three infrared LEDs positioned atop the microdrive EIB. Cineplex Editor (Plexon) was employed to enter event markers and to verify animal positional tracking data.
5
Open Field Habituation Locomotor Assessment
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Open field habituation task was performed in a 40 × 40 × 40 cm3 box during 2 × 15-min habituation sessions, spaced 1–2 h apart. Animals at 8–12 weeks of age were placed in the behavior box and allowed to explore freely during each habituation session. Sessions were recorded and locomotor activity was tracked using Plexon’s CinePlex Studio and analyzed using Plexon’s CinePlex Editor and code written in MATLAB. For analysis, the box was divided into two zones: an ‘inner’ zone (containing the inner 30 × 30 cm2 center square) and an ‘outer’ zone (the outermost area 5 cm from the walls). Total distance traveled was computed as the sum of distance covered over the course of each habituation session.
6
Extracellular Recordings and Position Tracking
7
Open Field Assay for Locomotor and Anxiety
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A standard open field arena (43 × 43 cm; Med Associates) was used to assess locomotor activity and anxiety-like behaviour. Seven days following vehicle or Fos ecRNA ASO infusions into the CA1 of the hippocampus, rats were placed in the open field arena for a 30-min period. Activity was tracked using automated video tracking software (CinePlex Studio, Plexon Inc.). Distance travelled (in cm) was used to quantify total locomotor activity, and time spent in the center of the open field (defined as 18 cm square in the middle of the field) was used to quantify anxiety-like behaviour.
8
Contextual Fear Conditioning and Open Field Exploration
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Contextual fear conditioning (CFC) was conducted as described [25 (link)]. Rats were placed into the training chamber with a metal floor grid (Med Associates) and allowed to explore for 7 min, during which three electric shocks (1 s, 0.5 mA each) were administered every 2 min. Memory was tested at 1 h, 24 h, and 7 days after training. An open field arena (43 × 43 cm; Med Associates) was used to assess locomotor and anxiety behavior, as previously described [25 (link)]. One week following CFC, rats were placed in the open field arena and allowed to explore for 30 min. Distance traveled (in cm) and time spent in the center (s) were tracked and quantified using automated video tracking software (CinePlex Studio, Plexon Inc).
Additional details for all procedures can be found insupplemental methods .
Additional details for all procedures can be found in
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