To evaluate whether the observer’s behaviour mainly vary because of the demonstrator mice’s hyperactivity or immobility we have used drugs for demonstrators. We investigated whether immobility times among observer mice decreased when demonstrator mice exhibited excessive activity. We treated the demonstrator mice, with MK-801 to reduce the immobility. In the present study, (+)-MK-801 (dizocilpine hydrogen maleate; 130-17381, FUJIFILM Wako Pure Chemical Corporation) was diluted in saline at a concentration of 0.1 mg/ml. Mice received an intraperitoneal (IP) injection of MK-801 of 0.2 mg/kg 30 min before being tested. The demonstrator mice were treated with pentobarbital to induce sleep and increase their immobility. Cagemate mice were deeply anaesthetised via a high dose of sodium pentobarbital (50 mg/kg, IP; Somnopentyl®, Kyoritsu Seiyaku Corp. Tokyo, Japan). The experimental apparatus consisted of two identical chambers (18 cm × 20 cm × 40 cm) with a transparent divider in the middle and a metal grid floor. Treated mice were placed in the demonstrator chamber, while cagemate mice were placed in the observer chamber (Fig. 2 a). The resultant behaviour was recorded for 4 min. Images were captured using a video camera, and immobility time was measured and evaluated using the ANY-MAZE software.
Mk801
Manufactured by Fujifilm
Sourced in Japan
The MK801 is a compact and versatile lab equipment designed for diverse applications in scientific research and analysis. Its core function is to provide accurate and reliable measurements across a wide range of parameters, supporting various laboratory workflows. The detailed specifications and intended use of this product are not available within the scope of this request.
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Lab products found in correlation
S 4 carboxyphenylglycine cpg, by Bio-Techne (3 mentions)
Muscimol, by Fujifilm (2 mentions)
Ly341495, by Bio-Techne (2 mentions)
N acetyl l cysteine nac, by Fujifilm (2 mentions)
Somnopentyl, by Kyoritsu Seiyaku (1 mentions)
Way100635, by Cosmo Bio (1 mentions)
Bp554, by Fujifilm (1 mentions)
Rs α cyclopropyl 4 phosphonophenyl glycine, by Bio-Techne (1 mentions)
Rs α cyclopropyl 4 phosphonophenyl glycine cppg, by Bio-Techne (1 mentions)
Clozapine, by Merck Group (1 mentions)
Haloperidol, by Fujifilm (1 mentions)
9 protocols using mk801
1
Evaluating Observer Behavior through Demonstrator Modulation
2
Pharmacological Modulation of Neurotransmitter Systems
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Lurasidone, the 5-HT1AR agonist BP554, the 5-HT7R antagonist SB269970, the GABAA-R agonist muscimol, and the noncompetitive NMDA-R antagonist MK801 were obtained from Fujifilm-Wako (Osaka, Japan). The 5-HT1AR antagonist WAY100635 and the 5-HT7R agonist AS19 were purchased from Cosmo-Bio (Tokyo, Japan).
All compounds were prepared on the day of experiments. All drugs were perfused in MRS containing 145 mM Na+, 2.7 mM K+, 1.2 mM Ca2+, 1.0 mM Mg2+, and 154.4 mM Cl−, which was adjusted to pH 7.4 using 2 mM phosphate buffer and 1.1 mM Tris buffer [7 (link),50 (link)]. muscimol, MK-801, WAY100635, and SB269970 were dissolved in MRS directly, whereas BP554 and AS19 were initially dissolved in dimethyl sulfoxide at 50 mM. Lurasidone was initially dissolved in dimethyl sulfoxide at 1 mg/mL. The final dimethyl sulfoxide concentration was lower than 0.1% (vol/vol).
All compounds were prepared on the day of experiments. All drugs were perfused in MRS containing 145 mM Na+, 2.7 mM K+, 1.2 mM Ca2+, 1.0 mM Mg2+, and 154.4 mM Cl−, which was adjusted to pH 7.4 using 2 mM phosphate buffer and 1.1 mM Tris buffer [7 (link),50 (link)]. muscimol, MK-801, WAY100635, and SB269970 were dissolved in MRS directly, whereas BP554 and AS19 were initially dissolved in dimethyl sulfoxide at 50 mM. Lurasidone was initially dissolved in dimethyl sulfoxide at 1 mg/mL. The final dimethyl sulfoxide concentration was lower than 0.1% (vol/vol).
3
Pharmacological Modulation of Neuronal Signaling
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NMDAR antagonist, MK801,15 (link) memantine (MEM), cysteine prodrug, N‐acetyl‐l ‐cysteine (NAC),16 (link) and the GABAA receptor agonist, muscimol (MUS)15 (link) were obtained from Wako Chemicals (Osaka, Japan). The II‐mGluR antagonist LY341495,17 (link) the III‐mGluR antagonist (RS)‐α‐cyclopropyl‐4‐phosphonophenyl glycine (CPPG),17 (link) and the Sxc inhibitor (S)‐4‐carboxyphenylglycine (CPG)18 (link) were purchased from Tocris Bioscience (Bristol, UK).
All compounds were prepared on the day of experiments. MK801, MEM, CPPG, CPG, NAC, and MUS were dissolved in modified Ringer's solution (MRS) or artificial cerebrospinal fluid (ACSF). LY341495 was initially dissolved in 10 mmol L−1 dimethyl sulfoxide and was then diluted to 1 mmol L−1 in MRS. The final concentrations of LY341495 and dimethyl sulfoxide were 1 μmol L−1 and 0.1% (vol vol−1), respectively.
MRS contained 145 mmol L−1 Na+, 2.7 mmol L−1 K+, 1.2 mmol L−1 Ca2+, 1.0 mmol L−1 Mg2+, and 154.4 mmol L−1 Cl−. The pH was adjusted to 7.4 using 2 mmol L−1 phosphate buffer and 1.1 mmol L−1 Tris buffer.19 (link), 20 (link) ACSF contained 130 mmol L−1 NaCl, 5.4 mmol L−1 KCl, 1.8 mmol L−1 CaCl2, 1 mmol L−1 MgCl2, and 5.5 mmol L−1 glucose and was buffered to pH 7.3 using 20 mmol L−1 HEPES.
All compounds were prepared on the day of experiments. MK801, MEM, CPPG, CPG, NAC, and MUS were dissolved in modified Ringer's solution (MRS) or artificial cerebrospinal fluid (ACSF). LY341495 was initially dissolved in 10 mmol L−1 dimethyl sulfoxide and was then diluted to 1 mmol L−1 in MRS. The final concentrations of LY341495 and dimethyl sulfoxide were 1 μmol L−1 and 0.1% (vol vol−1), respectively.
MRS contained 145 mmol L−1 Na+, 2.7 mmol L−1 K+, 1.2 mmol L−1 Ca2+, 1.0 mmol L−1 Mg2+, and 154.4 mmol L−1 Cl−. The pH was adjusted to 7.4 using 2 mmol L−1 phosphate buffer and 1.1 mmol L−1 Tris buffer.19 (link), 20 (link) ACSF contained 130 mmol L−1 NaCl, 5.4 mmol L−1 KCl, 1.8 mmol L−1 CaCl2, 1 mmol L−1 MgCl2, and 5.5 mmol L−1 glucose and was buffered to pH 7.3 using 20 mmol L−1 HEPES.
4
Pharmacological Modulation of Neurochemical Pathways
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Clozapine was purchased from Sigma (St. Louis, MO, USA). The non-competitive NMDA-R antagonist, MK801 [40 (link)], was obtained from Wako Chemicals (Osaka, Japan). The II-mGluR antagonist (LY341495), the III-mGluR antagonist ((RS)-α-cyclopropyl-4-phosphonophenyl glycine (CPPG)) [41 (link)], and the Sxc inhibitor ((S)-4-carboxyphenylglycine (CPG)) [30 (link),42 (link)] were purchased from Tocris Bioscience (Bristol, UK). The hemichannel and gap-junction blocker, carbenoxolone (CBX) [43 (link)], was obtained from Cosmo Bio (Tokyo, Japan).
LY341495 and CBX were initially dissolved in 10 mM with dimethyl sulfoxide, then diluted to 1 mM with modified Ringer’s solution (MRS) or artificial cerebrospinal fluid (ACSF) [28 (link),44 (link)]. CPPG and CPG were dissolved in MRS or ACSF. CLZ and MK801 were dissolved in MRS, ACSF, or Dulbecco’s modified Eagle’s medium containing 10% fetal calf serum (fDMEM) containing less than 0.1% acetic acid.
LY341495 and CBX were initially dissolved in 10 mM with dimethyl sulfoxide, then diluted to 1 mM with modified Ringer’s solution (MRS) or artificial cerebrospinal fluid (ACSF) [28 (link),44 (link)]. CPPG and CPG were dissolved in MRS or ACSF. CLZ and MK801 were dissolved in MRS, ACSF, or Dulbecco’s modified Eagle’s medium containing 10% fetal calf serum (fDMEM) containing less than 0.1% acetic acid.
5
Pharmacological Modulation of Motor Cortical LTP
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A separate group of animals were prepared to explore the correlation between motor cortical LTP induction after QTS5, QPS5, and OPS2.5 and NMDA-receptor activation, with rats receiving an NMDA-receptor antagonist, MK-801 (1 mg/kg, intraperitoneally; FUJIFILM Wako Pure Chemical Corporation, Japan; [B] in Figure 1 , n = 15). Low-dose MK-801 (0.5 mg/kg) was tested only for MEPs and CS preceding TS-MEP recording (n = 5). Based on the previously reported pharmacokinetic and pharmacodynamic properties of MK-801 in rats, we adjusted motor cortical LTP induction periods to make them comparable with the bet time window (Wegener et al., 2011 (link)). Furthermore, to confirm whether MK-801 was at a steady state during the induction of motor cortical LTP by QTS5, QPS5, and OPS2.5, the pharmacodynamics of MK-801 were tested by means of functional observation battery (FOB) using eight separate group of animals. We verified the positive steady symptoms during 1-h-post-dose periods (see Supplementary Material for detail). Thus, we could confirm that MK-801 doses were steady during the induction of LTP, but interaction with anesthetic combination was not confirmed.
6
Pharmacological Modulation of Glutamate Signaling
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The NMDA-R antagonists, MK801 and amantadine (AMA) [17 (link)], and a cysteine prodrug, N-acetyl-L-cysteine (NAC) [18 (link)] were obtained from Wako Chemicals (Osaka, Japan). The cystine/glutamate antiporter (Sxc) inhibitor (S)-4-carboxyphenylglycine (CPG) [19 (link)] was purchased from Tocris Bioscience (Bristol, UK). Selective group I metabotropic glutamate receptors (I-mGluRs) antagonist, YM298198, was purchased from Sigma-Aldrich (St. Louis, MO, USA). A selective AMPA-R antagonist, perampanel (PER), was obtained from Cosmo Bio (Tokyo, Japan). All compounds were prepared on the day of their use in experiments. MK801, AMA, YM298198 and NAC were dissolved in modified Ringer’s solution (MRS) or artificial cerebrospinal fluid (ACSF). PER was initially dissolved at a concentration of 1 mM in dimethyl sulfoxide. CPG was initially dissolved in 1 N NaOH and was diluted to 1 μM in MRS.
7
Preparation of L-PA and MK-801 Solution
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L-PA and MK-801 were purchased from Wako Pure Chemical Industries (Osaka, Japan) and were dissolved in 0.85% saline containing 0.1% Evans Blue solution. The suspension was stirred using a vortex mixer and sonicated using a bath sonicator for 10 min.
8
Pharmacological Modulation of Neural Circuits
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The noncompetitive NMDAR antagonist, MK801 (dizocilpine) and the GABAA-R agonist, muscimol, were obtained from Fujifilm-Wako (Osaka, Japan). The AMPAR antagonist, perampanel was obtained from Cosmo Bio (Tokyo, Japan). These compounds were prepared on the day of experiments. These drugs were perfused in MRS containing 145 mM Na+, 2.7 mM K+, 1.2 mM Ca2+, 1.0 mM Mg2+, and 154.4 mM Cl−, which was adjusted to pH 7.4 using 2 mM phosphate buffer and 1.1 mM Tris buffer [15 (link),16 (link)]. muscimol and MK801 were dissolved in MRS directly. Perampanel was initially dissolved at a concentration of 1 mM in dimethyl sulfoxide.
According to previous reports, in the present study, to study effects of MK801, muscimol and perampanel, each rat was administrated by perfusion with MK801 (1, 5 and 50 μM) [15 (link),16 (link),17 (link),20 (link),21 (link)], muscimol (1 μM) [23 (link),24 (link)] and perampanel (1 μM) [19 (link),20 (link)] into the mPFC, RTN or DRN.
According to previous reports, in the present study, to study effects of MK801, muscimol and perampanel, each rat was administrated by perfusion with MK801 (1, 5 and 50 μM) [15 (link),16 (link),17 (link),20 (link),21 (link)], muscimol (1 μM) [23 (link),24 (link)] and perampanel (1 μM) [19 (link),20 (link)] into the mPFC, RTN or DRN.
9
Pharmacological Modulation of Schizophrenia-Related Behaviors
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(+)-MK-801 (130–17,381), Memantine (131–18,313), and Haloperidol (084–04261) were purchased from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan) and Betahistine (B1424) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). All reagents were diluted in saline and administered intraperitoneally. (+)-MK-801 was diluted to a concentration of 0.1 mg/mL and administered at a dose of 0.2 mg/kg. This dose was selected based on previous studies demonstrating a schizophrenia-related deficient effect of MK-801 at 0.2 mg/kg in mice [43 (link)–45 (link)]. Memantine was diluted at a concentration of 10 mg/mL and administered at a dose of 20 mg/kg. This dose was selected based on previous studies demonstrating a hyperlocomotive effect of memantine at 20 mg/kg in mice [35 (link)]. Haloperidol was dissolved in saline with a minimal amount of acetic acid [46 (link)] and administered at a dose of 2 mg/kg, whereas, betahistine was diluted at a concentration of 10 mg/mL and administered at a dose of 10 mg/kg [35 (link)]. The same amount of acetic acid was added to the vehicle for control animals.
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