Figure 2.

Maternal programming leads to microglia activation and c-Fos response in hypothalamus. Offspring was programmed by CAF diet exposure as previously described, and 0.2 micrograms/kg ghrelin was intradermically administered, and subjects were intracardially perfused with 0.1M PBS + heparin followed by PBS + 4% PFA. Hypothalamic sections were obtained using a cryostat, according to the Paxinos and Watson Atlas. Immunofluorescence to identify microglia activation was performed using the Iba-1 antibody (1:200) following by the secondary antibody Alexa Fluor 546 Goat Anti-Rabbit (IgG) (1:1000) (a). c-Fos activation was identified by anti c-Fos (1:1000) and Alexa Fluor 488 Rabbit Goat Anti-Rabbit (1:1000) (b). Brain sections were mounted using Vectashield with DAPI (Vector Laboratories) on coverslips. (n = 3). PBS, phosphate-buffered saline, PFA, paraformaldehyde.