Brain derived neurotrophic factor (bdnf)

Human Aβ_1–42 was purchased from EMD Millipore Corporation (Billerica, MA, USA). Ori was obtained from Chengdu Must Bio-Technology Company (Sichuan, China). Neurobasal media and B27 were bought from Life Technologies Corporation. MAP-2 antibody was purchased from Abcam (Cambridge, MA, USA). COX-2 antibody was bought from Santa Cruz Biotechnology (Santa Cruz, USA). PSD-95 antibody, synaptophysin antibody and CREB antibody were obtained from CST (Cell Signaling Technology, USA). 4', 6'-diamidino-2-phenylindole (DAPI), VDAC1, BDNF, p-TrkB, TrkB, p-CREB and β-actin antibodies were purchased from Bioworld Technology (Bioworld, USA). Lamin B1 antibody and HRP-conjugated secondary antibodies were also obtained from Bioworld.

Recombinant human BDNF was purchased from PeproTech (Rocky Hill, NJ, USA). FSH (human pituitary) and protein G agarose were from Merck Millipore (Darmstadt, Germany). Hoechst 33342 for DNA staining was obtained from Sigma-Aldrich (Darmstadt, Germany). The following antibodies were used: rabbit polyclonal antibodies against β-actin, FSHR and BDNF (Bioworld Technology, Louis Park, MN, USA); rabbit monoclonal antibodies against CREB and phospho-CREB (Ser133) (Cell signaling Technology, Danvers, MA, USA); rabbit polyclonal antibody against ubiquitin and aromatase, and mouse monoclonal antibody against phosphoserine/threonine/tyrosine (Abcam, Shanghai, CHN). The following kits were employed: Human BDNF ELISA Kit (Boster, Wuhan, CHN); cAMP assay kit (R&D Systems, Inc., Minneapolis, MN, USA); PKA kinase activity kit (Abcam, Shanghai, CHN).