The resin-bound fractions were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and cellular GTP-Rap1 levels were analyzed by immunoblotting with anti-Rap1-GTP and anti-rap1, respectively. The band intensities were then normalized to anti-Rap1 in the input lane (defined as 1.0). In this study, 8-pCPT-2′-O-Me-cAMP-AM (BioLog Life Science Institute, Bremen, Germany) was dissolved in a vehicle solution containing 0.1% DMSO and 0.1%BSA. The sections from the CA1 stratum radiatum of the hippocampus of three male EPAC−/− at

96 days old of age and three control littermates (EPAC+/+ mice) were studied. Mice were perfused with 2% paraformaldehyde + 2% glutaraldehyde in 0.1 M phosphate buffer, then postfixed in 1% osmium tetroxide in 0.1 M cacodylate Androgen Receptor antagonist buffer, stained en bloc with 1% uranyl acetate in 50% ethanol, dehydrated in an ethanol series and then put in propylene oxide and embedded in epon. Thin sections were stained with lead citrate.

Golgi staining was performed on four strains of male mutant mice and their respective wild-type controls at 90 ± 5 days old of age. In ALK signaling pathway this study, FD rapid Golgi Stain kit (FD NeuroTechnologies,) was used. Briefly, brains were removed from mice and immediately immersed in solution A and B for 2 weeks at room temperature and transferred into solution C for 24 hr at 4°C, as instructed on the manufacture’s experimental methods. The brains were sliced using a Vibratome (VT1000S; Leica) at a thickness of 100 μm. Bright-field microscopy (Axio Observer; Zeiss) images were taken of CA1 pyramidal neurons (80 cell with a total of 80 cm length of dendrites per group were analyzed). Images 4-Aminobutyrate aminotransferase were coded and synaptic spines counted in software with Image Probes. All the spines counted were also measured for spine length and spine densities were expressed as spine/μm dendrite. Comparisons between genotypes were carried out using two-way ANOVA. Electrophysiological experiments and biochemical assays were analyzed using

the Student t test. LTP analysis was performed on 10 min blocks of data within the last 30 min of the recording. This work was supported by National Natural Science Foundation of China (Grant 81130079 YL), National Institute of Health (NIH/NINDS, R01NS5051383Y.L and NIH/NIA R01AG033282Y.L), the New Century Excellent Talents in University (NCET-10-0421, L.-Q.Z.), and the NIH/NIDCD Intramural Program (R.S.P. and Y.-X.W.). “
“There is abundant evidence demonstrating a key role for the hippocampus and MEC in landmark- and path integration-based navigation (O’Keefe and Nadel, 1978, Morris et al., 1982, Nadel, 1991, McNaughton et al., 1996, McNaughton et al., 2006, Whishaw et al., 2001a, Parron and Save, 2004 and Steffenach et al., 2005). Both areas contribute to spatial mapping, with place cells in the hippocampus firing at particular locations in the environment (O’Keefe and Dostrovsky, 1971), and grid cells in MEC providing a precise two-dimensional metric for space (Hafting et al.