Cytosolic Phospholipase A2 Facilitates Oligomeric Amyloid-β Peptide Association with Microglia via Regulation of Membrane-Cytoskeleton Connectivity.

Cytosolic phospholipase A₂ (cPLA₂) mediates oligomeric amyloid-β peptide (oAβ)-induced oxidative and inflammatory responses in glial cells. Increased activity of cPLA₂ has been implicated in the neuropathology of Alzheimer's disease (AD), suggesting that cPLA₂ regulation of oAβ-induced microglial activation may play a role in the AD pathology. We demonstrate that LPS, IFNγ, and oAβ increased phosphorylated cPLA₂ (p-cPLA₂) in immortalized mouse microglia (BV2). Aβ association with primary rat microglia and BV2 cells, possibly via membrane-binding and/or intracellular deposition, presumably indicative of microglia-mediated clearance of the peptide, was reduced by inhibition of cPLA₂. However, cPLA₂ inhibition did not affect the depletion of this associated Aβ when cells were washed and incubated in a fresh medium after oAβ treatment. Since the depletion was abrogated by NH₄Cl, a lysosomal inhibitor, these results suggested that cPLA₂ was not involved in the degradation of the associated Aβ. To further dissect the effects of cPLA₂ on microglia cell membranes, atomic force microscopy (AFM) was used to determine endocytic activity. The force for membrane tether formation (F_mtf) is a measure of membrane-cytoskeleton connectivity and represents a mechanical barrier to endocytic vesicle formation. Inhibition of cPLA₂ increased F_mtf in both unstimulated BV2 cells and cells stimulated with LPS + IFNγ. Thus, increasing p-cPLA₂ would decrease F_mtf, thereby increasing endocytosis. These results suggest a role of cPLA₂ activation in facilitating oAβ endocytosis by microglial cells through regulation of the membrane-cytoskeleton connectivity. [ABSTRACT FROM AUTHOR]