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The mice, under isoflurane anesthesia (1% at 2 l/min oxygen flow) for at least 30 min, were placed in prone position on the bed of a LabPET™ small animal PET scanner (Gamma Medica, Northridge, CA, USA), having a 3.75-cm axial field of view and achieving 1.35-mm resolution [24].
Twenty-four h later the animals received the microinjections and after 10 min were placed into the test box.
PMA (Biotium, Hayward, CA, USA) was added to the samples at a final concentration of 50 μM and the samples shaken in the dark for 5 min were placed in the PMA lamp apparatus (LED-Active Blue, Ingenia Biosystems, Barcelona, Spain) for 15 min.
The mice that reached the platform in <1 min were placed on the center of the elevated dowel at 30 cm from the platform, and the latency time to reach the platform, the number of arrivals, and the number of falls were recorded during 90 s.
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This operation (modifying the part of the matrix where CRM min is placed, in order to improve its value) changes GRM(H).
A 1 mm burr hole was placed 3 mm lateral to the bregma and a stereotactic implantation of 3 µl cell suspension injected over 3 min was placed in an area corresponding to the internal capsule 0.5 mm below the tracts of the corpus callosum.
A time limit of 10 min was placed on each session.
The probe, denatured at 75°C for 5 min, was placed on the denatured slides, covered with Parafilm (American National Can, Greenwich, CT), and incubated in a humid box at 37°C overnight.
After 2 min, mice were placed back into the arena, and left to explore for 5 min. Mice were then removed from the arena and returned to the home cage.
After fixation with 1 % formaldehyde for 10 min metaphases were placed in denaturing solution (70%% formamide in 2 x SSC) at 72 °C for 7 min followed by dehydration.
After 1 min, oocytes were placed in 1 ml solution containing 0.5 M sucrose, and incubated at RT for 3 min. Finally, the oocytes were washed at RT for 6 min in two different dishes containing 1 ml basic medium, then transferred into 1 ml culture media (Cleavage media, Sage).
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