Immunohistochemical Analysis of Rat Nervous System

Five rats of each parental strain BN and SHR were used for immunohistochemistry and minimum three sections per rat were evaluated from each tissue; whiskers area, trigeminal ganglia and brain stem. The method used for tissue preparation, staining and antibodies were described previously [8 (link)]. The antibodies used include polyclonal rabbit anti-HSV-1 (1:100) (Dako), mouse monoclonal anti-Tuj1 (1:500) (Covance), mouse monoclonal anti-ED1 (1:200) (Serotec), mouse monoclonal anti-NKRp1 (1:200) (Harlan Sea-Lab) and mouse monoclonal anti-CD8 (1:200) (Serotec). Additional antibodies used in this study include; polyclonal rabbit anti-von Willebrand factor Ab (1:200) (Abcam) to visualize vWF in the endothelial cell layer and mouse monoclonal anti-Occludin 1A8 Ab (1:200) (Antibodies Online) to visualize the expression of tight junction proteins. After incubation with primary antibodies, sections were rinsed and incubated with Alexa Fluor^™®488 goat anti-rabbit (1:200) (Molecular Probes, USA) or Alexa Fluor^™® 594 goat anti-mouse (1:200) (Molecular Probes) secondary antibodies. Micrographs were taken on a Zeiss Axioskop microscope system and processed in Adobe Photoshop CS3. The pictures included in the Figs represent the staining differences visualised in all slides from the different compartments of all the rats studied.