A Tat-conjugated peptide, designed to block the binding of dynamin to amphiphysin and thus prevent endocytosis (Gout et al., 1993 (link); Lissin et al., 1998 (link); Kittler et al., 2000 (link); Lin et al., 2011 (link)), was infused in order to block endocytosis. Depending on the experiment, rats received bilateral infusions of Tat P4 peptide and Tat Scrambled control peptide (S) (60μg/μl/0.5 μl side; Cambridge, UK), human recombinant BDNF or saline (0.5 μg/μl/0.5 μl side), ANA-12 or saline at different times during the behavioral task. The injection volume was always 0.5 μl/side. Sequences are as follows: amino acid sequence for the dynamin inhibitory peptide (P4) is QVPSRPNRAP, and for the Scrambled control peptide (S) is QPPASNPRVR.
Bilateral infusions were conducted simultaneously using two 5-μl Hamilton syringes that were connected to the infusion cannulas by propylene tubing. Syringes were driven by a Harvard Apparatus precision syringe pump, which delivered 0.5 μl to each hemisphere over 1 min. The infusion cannulas were left in place for an additional minute to allow for diffusion. At least 3 days were allowed for washout between repeated infusions.
Bilateral infusions were conducted simultaneously using two 5-μl Hamilton syringes that were connected to the infusion cannulas by propylene tubing. Syringes were driven by a Harvard Apparatus precision syringe pump, which delivered 0.5 μl to each hemisphere over 1 min. The infusion cannulas were left in place for an additional minute to allow for diffusion. At least 3 days were allowed for washout between repeated infusions.
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