Effect of Acetylcholine Precursors on Proliferation and Differentiation of Astroglial Cells in Primary Cultures

Abstract

Effects of acetylcholine and of the cholinergic precursors choline, cytidine 5'-diphosphocholine (CDP-choline) and alpha-glyceryl-phosphorylcholine (alpha-GPC) on transglutaminase (TG) and cyclin D1 expression were studied in primary astrocyte cultures by confocal laser microscopy (CLSM) with monodansyl-cadaverine uptake as a marker of enzyme activity and by immunochemistry (Western blotting). CLSM analysis showed an increased cytofluorescence in 0.1 microM choline-treated astrocytes. Treatment with CDP-choline dose-dependently increased TG. A total of 1 microM CDP-choline exposure in 14 days in vitro (DIV) astrocyte cultures increased cytofluorescence. A total of 1 microM alpha-GPC 24 h-treated cultures revealed increased cytofluorescence both in cytosol and nuclei. Western blot analysis showed an increased TG expression in cultures exposed for 24 h to 1 microM choline or alpha-GPC, whereas in 24 h 1 microM CDP-choline and acetylcholine-treated astrocytes TG expression was unaffected. Treatment with 1 microM acetylcholine reduced TG expression at 21 DIV. In cultures at 14 and 35 DIV cholinergic precursor treatment for 24 h induced a marked down-regulation of cyclin D1 expression, with reduced cyclin D1 expression in 1 microM alpha-GPC treated astrocytes. Our data suggest a role of cholinergic precursors investigated independent from acetylcholine on maturation and differentiation of astroglial cells in vitro, rather than on their growth, proliferation and development in culture.

The Role of Citicoline in Neuroprotection and Neurorepair in Ischemic Stroke

“…The diagram explains the actions of citicoline to enhance the processes of inhibition of apoptosis [48], angiogenesis [92], neurogenesis [84], gliagenesis [93], synaptogenesis [50], and modulation of neurotransmitters [84]. Notice that all these effects are similar to those induced by stem cells.…”

“…Long-term treatment with citicoline is safe and effective, improving post-stroke cognitive decline and enhancing patients’ functional recovery. Prolonged citicoline administration at optimal doses has been demonstrated to be remarkably well tolerated and to enhance endogenous mechanisms of neurogenesis and neurorepair, similar to those obtained with stem cells [92,93], contributing to physical therapy and rehabilitation.…”

Serum deprivation alters lipid profile in HN9.10e embryonic hippocampal cells

“…Furthermore, hydrolysis of PC by phospholipase D is a source of choline, which is not only used for the synthesis of the neurotransmitter acetylcholine in cholinergic neurons [7], but it is possibly involved in maturation and differentiation of astroglial cells in vitro independently of acetylcholine [8,9]. The multiple pathways of ceramide generation led to the hypothesis that individual ceramide molecular species are regulated by specific biochemical pathways in distinct subcellular compartments and they carry out distinct functions [10].…”

Effect of Scopolamine-based Amnesia on the Number of Astrocytes in the Rat's Hippocampus

“…Treatment with a-GPC reduce glial reaction levels in the hippocampus that it is possible to cause neuronal protection and astroglial dynamic effects and some important neurodegenerative disorders such as brain aging, stroke, ischemic brain disease Parkinson's and Alzheimer's to be effective (Bramanti et al, 2008).…”