Temporal and sequential analysis of microglia in the substantia nigra following medial forebrain bundle axotomy in rat

Abstract

Dopaminergic neurons in the substantia nigra pars compacta undergo apoptosis after transection of the medial forebrain bundle. We have assessed the temporal and sequential activities of microglia in these events by examining the complement-3 (OX-42), major histocompatibility complex class II antigen presentation (OX-6) and phagocytic activity (ED1), and correlating these indicators with dopaminergic neuronal loss. Microglia in the ipsilateral substantia nigra pars reticulata evinced activation morphology at 12 h postaxotomy. Phagocytic microglia apposed dying dopaminergic neurons in the pars compacta starting at 3 days postlesion; their number increased through 14 days and slowly decreased. Nuclear chromatin condensation and significant loss of tyrosine hydroxylase-positive dopaminergic neurons occurred around 7 days postlesion. In contrast to microglial expression of interleukin-1β and inducible nitric oxide synthase at the axotomy site, nigral microglia were interleukin-1β and inducible nitric oxide synthase-negative. Consistently, RNase protection assays showed that interleukin-1β and inducible nitric oxide synthase transcripts in nigra were equivocal. The present data support the idea that phagocytosis of axotomized neurons by activated microglia is not limited to dead neurons but includes dying neurons probably without cytotoxic effects of inflammatory substances, such as interleukin-1β or nitric oxide.

Keywords microglia; dopaminergic neurons; Parkinson’s disease; phagocytosis; neurodegenerative disease; apoptosis