Labeling of amoeboid microglial cells and intraventricular macrophages in fetal rats following a maternal injection of a fluorescent dye

Abstract

Amoeboid microglial cells (AMC) in fetal brains were labeled by rhodamine B isothiocyanate (RhIc) when injected intravenously or intraperitoneally into mother rats at late stage of pregnancy. The fluorescent cells were immunostained with antibodies OX-42 and OX-18 that recognize complement type 3 (CR3) receptors and major histocompatibility complex class I (MHC-I) surface antigen, respectively. RhIc-labeled AMC were first observed in the cavum septum pellucidum and subependymal cysts associated with the cerebral aqueduct as well as the fourth ventricle, and subsequently at other sites including the corpus callosum and other subcortical white matter. The fluorescence intensity increased with time after RhIc administration so that after 1 day the cells were brightly labeled. The majority of the labeled cells were round, with some elongated ones bearing two or three processes. Besides AMC, macrophages in the ventricular system were also labeled. All fluorescent cells were double labeled with OX-42 and OX-18 antibodies. Present results suggest that when introduced into the maternal circulation, RhIc could readily gain access into the fetal brain through the inefficient placental, blood-brain and blood-cerebrospinal-fluid (blood-CSF) barriers. The avid uptake of RhIc in circulation by brain macrophages indicates an active scavenging role of these cells in fetal brian. The labeling of cells by maternal route offers a rapid method for study of distribution of brain macrophages in fetuses.