Behavioral experiments were conducted in a sound-attenuated and air-regulated room, where the animals were habituated 1 h prior to experiments. All animal experimentation reported in this study was performed under established standards of the Brazilian law No. 11.794/2008 in accordance with the Policies on the Use

of Animals and Humans in Neuroscience Research, revised and approved by the Society for Neuroscience Research. Purification of Tx3-1 from the venom of the spider Phoneutria nigriventer was performed following the method of Cordeiro and coworkers (1993). Aβ25-35, learn more Aβ35-25 and 4-aminopyridine (4-AP), were purchased from Sigma (St. Louis, MO, USA). The Tx3-1 and 4-AP dose were based on electrophysiological experiments that evaluated the effect of both compounds on IA currents ( Kushmerick et al., 1999). Aβ aggregation was performed according to Maurice et al. (1996), wherein Dapagliflozin datasheet 3 mM of either 25-35 or 35-25 (used as control) sequence peptide were incubated at 37 °C for 4 days, stored at −20 °C and freshly diluted to the final dose (3 nmol/site; 1 mM) when used. In all behavioral experiments, Aβ25-35, Aβ35-25, Tx3-1, 4-AP or vehicle, were administered by intracerebroventricular

(i.c.v.) route, according to Laursen and Belknap (1986). Briefly, mice were anesthetized with isofluorane until full anesthesia was achieved. The microinjections were performed using a Hamilton 10 μl syringe connected to a specially made 28-gauge stainless steel needle with 3 mm in length. The needle was inserted directly through the skin and skull into the lateral

ventricle, targeted by visualizing an equilateral triangle between the eyes and center of skull to locate bregma, then inserting the needle 1 mm laterally to this point. This avoids the use of unnecessary force since the needle penetrates at the suture line of the skull plates. Compounds were injected in a volume of 5 μl over a 5 s period, followed by a 10 s delay to allow diffusion and prevent backflow. All injections were performed by an experimenter well trained in this technique. Novel object recognition task Chloroambucil was performed in wooden chamber (30 × 30 × 30 cm) with black side and rear walls, front wall made of transparent acrylic and the floor covered with an ethyl vinyl acetate sheet. A light bulb, hanging 60 cm above the behavioral apparatus, provided constant illumination of about 40 lux, and an air-conditioner provided constant background sound isolation. The objects used were plastic mounting bricks, each of them with different shapes and colors, but same size. Throughout the experiments objects were used in a counterbalanced manner and animals did not previously display preference for any of the objects. Chambers and objects were thoroughly cleaned with 30% ethanol before each experiment. Six days after Aβ injection, novel object recognition task was performed according to Wang and coworkers (Wang et al.