Fig. 6.

Effect of NT-3 and BDNF on total neuronal numbers and on dendritic morphology of excitatory neurons in E16 hippocampal cultures. A, At 14 d in vitro, cultures were fixed and immunostained for MAP2ab. To determine the number of cells, a total of 10 fields per coverslip were counted using 400× magnification under fluorescence filters or bright field (Zeiss Axioplan microscope). Results are expressed as the total number of cells staining for MAP2ab in 10 fields. Results are the mean ± SEM of data from seven cultures from four experiments.B–D, Cells were grown for 14 d in control conditions (CT) or from days 11–14 in the presence of NT-3 orBDNF. Cells were double immunostained for MAP2ab and GABA. MAP2ab-positive/GABA-negative neurons were traced with a camera lucida using a 63× objective, and from these drawings the number of primary dendrites and dendritic branches were directly counted. The total length of dendrites was measured with a digital plan measure and the NIH Image program. Results are the mean ± SEM of data from 10 cells per group from two experiments. CT, control;NT, NT-3 plus BDNF.