, 2007). See the Supplemental Data for details on the generation of Cxcr7flox/+ mice. See the Supplemental Data for details on immunohistochemistry and in situ hybridization. In utero electroporation with pCAGGS-Cxcl12 and pCAGGS-DsRed2 or pCAGGS alone was Sorafenib cost performed as described (Li et al., 2008). For blocking of CXCR4 receptors, 1 μl of AMD3100 solution (12.6 mM; Sigma) or PBS was injected into the lateral ventricle at E14.5. Coronal slices (200 μm) of E15.5 Cxcr4+/−

or Cxcr7+/− control and littermate Cxcr4−/−or Cxcr7−/− mutant brains were placed onto nucleopore membrane filters over 35 mm glass-bottom dishes containing Minimum Essential Medium (GIBCO) and 10% FBS. Mounted slices were immediately transferred to a 37°C/5% CO2 live tissue incubation chamber attached to a Zeiss inverted microscope and a PASCAL confocal laser scanning system and imaged repeatedly every 12 min for up to 20 hr. Real-time interneuronal migration patterns were quantified by using Zeiss LSM Image Browser software. See the Supplemental Data for details on the primary cultures and the transwell assay. For CXCR7 staining, the cells were fixed in 2% formaldehyde for 30 min at RT and then permeabilized with permeabilization solution (PBS/0.1% BSA/0.2% saponin) for 10 min at RT. Cells were incubated in blocking solution (PBS/1% BSA/0.1% saponin/1%

goat serum) for 30 min at RT. Cells were then incubated with CXCR7 antibody (11G8, 1:400; ChemoCentryx) in blocking solution for 30 min on ice. For double CHIR-99021 price labeling, cells were first incubated with CXCR7 antibody and then incubated with the other primary antibodies for 2 hr at RT. Statistical analysis was performed with Student’s t test, x2 test, or the one-way ANOVA and Tukey-Kramer post-hoc test using Prism4 (GraphPad Software) and XLSTAT (Addinsoft) software. p <0.05 was considered statistically significant. We

thank Marc von Zastrow and Mark Penfold for helpful discussions and reagents. This work was supported by the research grants to J.L.R.R. from Citizens United for Research in Epilepsy (C.U.R.E.), Nina Ireland, Larry L. Hillblom Foundation, Weston Havens Foundation, and NIMH R37 MH049428; to Y.W. from C.U.R.E Rhode Island Award from the Epilepsy Foundation; and to S.J.P. from K02 MH074985 and R01 MH077694. Timothy W. Behrens and Jason E. Long are full-time PDK4 employees of Genentech, Inc. Dianna Crawford is a full-time employee of Amgen, Inc. “
“Chemokines are a large family of small proteins that are characterized by their ability to induce chemotaxis in responsive cells. Several chemokines and their receptors are expressed in the developing CNS, among which Cxcl12 (also known as Stromal cell-derived factor-1, SDF1) is the most studied. Cxcl12 has been shown to promote the migration of granule cells in the cerebellum and hippocampus (Bagri et al., 2002, Ma et al., 1998, Zhu et al., 2002 and Zou et al., 1998), Cajal-Retzius cells (Borrell and Marín, 2006 and Paredes et al.