Immunohistochemical Analysis of Neuroendocrine Markers

Primary antibodies used are: rabbit anti-GnRH (Beauvillain and Tramu, 1980 (link)), sheep anti-KP AC053 (Franceschini et al., 2013 (link)), sheep anti-GnRH (Skrapits et al., 2015 (link)), mouse anti-p140Cap (Grasso et al., 2018 (link), see Supplementary Figure 1 for specificity test), guinea pig anti-Vesicular GABA Transporter (VGAT, Synaptic System, used 1:500), guinea pig anti-Vesicular glutamate Transporter (VGLUT, Synaptic System, used 1:500), rabbit anti-c-Fos (sc-52 Santacruz, used 1:500) and rabbit anti-cleaved caspase 3 (Cell Signaling Technology, used 1:400, Whittaker et al., 2021 (link)). Secondary antibodies used are: Alexa-Fluor 488 goat anti-rabbit (Invitrogen, used 1:400), Cy3 goat anti-rabbit (Invitrogen, used 1:800), Alexa-Fluor 568 goat anti-mouse (Invitrogen, used 1:400), FITC anti-guinea pig (Sigma F6261, used 1:100), Cy3 donkey anti-sheep (Jackson, used 1:1000). Peroxidase-conjugated secondary antibodies were obtained from GE Healthcare.