HSP27 expression was investigated in cultured neurons and glial cells isolated from fetal human brains using immunoblotting and immunocytochemistry. Under unstressed conditions, HSP27 was identified at a high level in astrocytes (> 99%), at a low level in neurons (7%), and at a minimally detectable level in microglia (< 1%), whereas it was undetectable in oligodendrocytes. Under these conditions, HSP27 was located in the cytoplasm, fractionated into the Triton X-100-soluble phase, and composed chiefly of the basic isoform (HSP27a). After exposure to heat stress (43 degrees C/90 min), the level of HSP27 expression was not altered in astrocytes but was elevated significantly in neurons (11-21%) and microglia (4-7%) during 8-48 hr postrecovery periods, while it remained undetectable in oligodendrocytes. In addition, various human neural cell lines exhibited differential patterns of HSP27 expression under unstressed and heat-stressed conditions. Following heat shock treatment (45 degrees C/30 min), granular aggregates of HSP27 were identified in the cytoplasm of astrocytes. Under heat-stressed conditions, HSP27 was distributed within the Triton X-100-insoluble fraction associated with an increase in two more acidic isoforms (HSP27b and HSP27c). HSP27 and alpha B-crystallin were coexpressed in astrocytes under unstressed and heat-stressed conditions. When astrocytes were exposed to known HSP27 inducers, hydrogen peroxide and cysteamine reduced the synthesis of HSP27, while estradiol showed no effects. The differential patterns of constitutive and heat-induced expression of HSP27 in cultured human neurons and glial cells suggest that the cellular mechanisms by which HSP27 expression is regulated are different among various cell types in the human central nervous system.