The endosomal trafficking of apolipoprotein E3 and E4 in cultured human brain neurons and astrocytes.

The association of the E4 isoform of apolipoprotein E (apoE) as a genetic risk factor for late onset Alzheimer's disease (AD) has been well established. Central nervous system (CNS) neurons are specifically affected so that defining the mechanisms by which two of the major human apoE isoforms act within CNS neurons is important to our understanding of their effect on neuronal maintenance and function. We have developed a cell culture model using human brain tissue to characterize exogenous apoE transport. We have tracked the association of apoE3 and E4 with CD63, the GTP-binding protein rab5a and the acidic hydrolase cathepsin D, which localize lysosomes, early endosomes, and late endosomes/lysosomes, respectively. Double immunostaining and confocal laser scanning microscopy revealed by z-series that after 30 min most intraneuronal apoE colocalized with rab5a, whereas no astrocyte apoE/rab5a colocalization was detected. Conversely, apoE3 and CD63 did not colocalize in neurons, even after 1 h, but was colocalized in astrocytes. Also, there was approximately 9% apoE3 colocalization with cathepsin D in neurons, whereas up to 87% of apoE4 vesicles were colocalized. In astrocytes, the proportion of apoE3 colocalized with cathepsin D was greater than that in neurons, but still significantly different from that found with apoE4. These immunohistological data demonstrate that, in neurons, apoE can be endocytosed via a rab5a-regulated vesicle-mediated pathway and that beyond this stage there may be isoform specific differences in apoE trafficking present in both neurons and astrocytes. Copyright 2001 Academic Press.