Regulation of the rat oligodendroglia cell line OLN‐93 by antisense transfection of butyrylcholinesterase

Abstract

Butyrylcholinesterase (BChE) is a glial cell marker with unknown function. For neuroepithelial cells, BChE has been shown to regulate cell division and expression of the postmitotic marker acetylcholinesterase (AChE), while similar studies are lacking for glial cells. By transducing an antisense‐5′BChE cDNA expression vector via calcium phosphate precipitation, we have analyzed the effect of BChE inhibition on proliferation and differentiation of rat oligodendroglia‐derived OLN‐93 cells. OLN‐93 cells were chosen because they are highly proliferative, while expressing markers of differentiated oligodendrocytes (Richter‐Landsberg and Heinrich, 1996). First, we established that OLN‐93 cells do express BChE protein, albeit chiefly in an inactive state, and that BChE was decreased by antisense‐5′BChE transfection. Cell proliferation was also strongly diminished, protein kinase C (PKCα) was upregulated, and expression of cytoskeletal and cell surface proteins was altered. In particular, immunoreactivities of the intermediate filament proteins vimentin and the cell adhesion protein F11 were detected, indicating that BChE‐inhibited OLN‐93 cells have shifted toward an astrocytic phenotype. These data support a role of the glia marker BChE in CNS glial cell proliferation and differentiation, achieved via a nonenzymatic mechanism. The possible biomedical impact of BChE protein, e.g., on CNS nerve regeneration, is briefly discussed. GLIA 31:195–205, 2000. © 2000 Wiley‐Liss, Inc.

PKC stimulation increases expression of Cholinesterases in R28 cell line

“…The surprising finding is that application of DOG along with BChE knockdown leads to more pronounced increase in PKC-α expression as compared with controls ( Fig 1). BChE knockdown leads to an increase in PKC-α expression (Fig 2), [7,8,14]. Here, we show that DOG stimulation along with BChE knockdown leads to an over-stimulation of PKC-α, adding an effect.…”

“…DOG is a diacyl glycerol analog of the classical stimulator of calcium-dependent PKC-α signaling. Previously, the interaction between ChE and PKC-α is shown [7,[8][9][10][11][12][13][14]. In this study, we stimulated embryonic cell line R28 with DOG under normal cell conditions as well as knocking BChE down.…”

“…Previously down-regulation of BChE leading to increased AChE and PKC-α expression along with the change in activation status of ERK pathway was displayed [8,14]. The effectors of change were found as transcription factor and activator associated with ERK/ PKC pathway, c-fos and P90RSK1 [8].…”

Lipid homeostasis and apolipoprotein E in the development and progression of Alzheimer's disease

“…Patients with apoE4 may be more vulnerable to developing white matter lesions in the presence of vascular risk factors such as hypertension (196). Thus, factors that mediate glial activation, proliferation, and maturation, including butyrylcholinesterase (BuChE), acetylcholinesterase (AChE), peroxisome proliferator-activated receptor activation, high peripheral cholesterol levels, injury, ischemia, EFAs and their derivatives, and A␤, may interact with apoE isoforms to mediate glia-induced inflammation and oxidative stress in the brain (93,191,194,(197)(198)(199). Chronic gliosis has been shown to trigger altered amyloid processing in vivo (200), and the pathological changes are associated with AD (201).…”

“…AChE, BuChE, and APOE all have roles in glial activation (191,197,198) and neurite outgrowth (146,218). Both AChE and APOE have roles in A␤ fibrilization (188,219), and BuChE and APOE have roles in lipid metabolism and transport (13,(220)(221)(222).…”