The development of microglia involves the expression of a phenotype displaying phagocytic behaviour termed brain macrophage or amoeboid microglial cell. We have previously shown that rat brain macrophages purified in vitro secrete thrombospondin, an extracellular matrix protein, which acts on cultured neuronal cells by promoting neurite growth. In the present study, the expression of thrombospondin was investigated in tissue sections of the developing rat forebrain in relation to the distribution of microglia. These cells were identified using anti-macrophage antibodies and the isolectin B4 from Bandeiraea simplicifolia. Immunocytochemical detection of thrombospondin clearly outlined a cell population displaying the morphologies and distribution of brain macrophages, from the 17th day of embryonic life up to the end of the second postnatal week. These cells were most numerous in cortical and subcortical regions of developing fibre tracts such as the corpus callosum or the internal capsule. The localization of thrombospondin in brain macrophages was confirmed by double immunostaining using ED1 monoclonal anti-macrophage antibodies. Ramified microglial cells were also labelled transiently by anti-thrombospondin antibodies during early postnatal life. These results provide in situ evidence supporting the notion that microglial cells could favour axonal growth by producing thrombospondin during development.