Microscopic slide

SYBR Green I nucleic acid stain, 10,000× concentrate was purchased from Invitrogen, USA, and a working solution of 10× SYBR Green I was prepared in Milli-Q water. p-Phenylenediamine and chloroform were obtained from Acros Organic (Fair Lawn, NJ, USA) and a 10% (wt/vol) stock solution of p-phenylenediamine was prepared in Milli-Q water. Whatman® anodisc inorganic filter membrane (13 mm, 0.02 μm pore size) was obtained from GE Healthcare (Buckinghamshire, UK). Microscopic slide was obtained from VWR international (Radnor, PA, USA). Tryptic soy broth (TSB) and tryptic soy agar (TSA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Filtration system, cover glass and Luria Bertani (LB) broth were obtained from Fisher Scientific (Pittsburgh, PA, USA). Phosphate buffered saline (PBS) was purchased from Fisher Bioreagents (Fair Lawn, NJ, USA). Polycarbonate filter (20 um pore size, 47 mm diameter) was purchased from Maine Manufacturing (ME, USA). Milli-Q water was produced by QPAK^® 2 purification system (EMD Millipore, Billerica, MA, USA).

Fourteen sections of 14 μm thickness were sectioned from each frozen hippocampal sample, and sections were collected onto polyethylene terephthalate metal frame slides for laser capture microdissection (Leica, Milton Keynes, UK). Two additional sections were collected onto a microscopic slide (VWR International, UK) and stained briefly in 0.1% toluidine blue (pH 4.5) solution for 5 s to visualize the granule cell layer. laser capture microdissection (LCM 700; Leica, Milton Keynes, UK) was then carried out along the entire length of the GCL of multiple sections per case to capture basal and dispersed DGCs (Figure 2A). Basal samples included DGCs in the granule cell layer closest to CA4, while the dispersed samples included ectopic DGCs in the outer-granular layer and inner and outer molecular layers. A total tissue area of 9 ± 1 mm² was dissected for each case, and submitted for proteomic analysis.