Immunocytochemical Localization of Fos in Perfused Nonhuman Primate Brain Tissue: Fixation and Antisera Selection

Abstract

Immunocytochemical localization of immediate early gene proteins, such as Fos, provides a powerful tool with which to demonstrate activated neuronal populations in response to specific stimuli. In contrast to studies using rat brain tissue that consistently show good Fos detection with a variety of antisera, studies using brain tissue from other species yield variable Fos detection. This may be partly due to differences in Fos protein sequences among species or to perfusion and fixation methods. To determine the ability of various Fos antisera to detect neuronal activation in nonhuman primate tissue, we tested nine Fos antisera and compared these antibodies under conditions of intense or physiological stimulation. Monkey brain tissue was either perfused and postfixed with 4% paraformaldehyde or perfused with 4% paraformaldehyde and postfixed with 2.5% acrolein in 4% paraformaldehyde. In rat tissue, stained for comparison, several antisera resulted in good to excellent Fos detection. However, few antisera tested in monkey tissue resulted in excellent Fos staining. We demonstrate that detection of Fos in monkey brain tissue perfused with 4% paraformaldehyde can be improved by postfixation in a dilute acrolein solution. Our findings emphasize the importance of choosing appropriate antisera and perfusion–fixation procedures to optimize Fos detection in nonhuman primate tissue.