Acetylcholine esterase is a regulator of GFAP expression and a target of dichlorvos in astrocytic differentiation of rat glioma C6 cells

Ozawa, Aisa; Kadowaki, Erina; Haga, Yosuke; Sekiguchi, Hitomi; Hemmi, Natsuko; Kaneko, Takuya; Maki, Takehiro; Sakabe, Kou; Hara, Shuntaro; Yamamoto, Masayuki; Arishima, Kazuyoshi; Sakaue, Motoharu

doi:10.1016/j.brainres.2013.08.031

Cited by 9 publications

(6 citation statements)

References 23 publications

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“…Upon differentiation with dBcAMP/theophylline C6 glioma cells have increased expression of the glial fibrillary acidic protein (GFAP). 217,218 Furthermore, after this differentiation protocol, cells present morphological alterations, including long projections or cellular processes. 217,218 Other agents are described to induce differentiation of this cell line.…”

Section: C6 Cells: a Rat Glioma Cell Linementioning

confidence: 99%

“…217,218 Furthermore, after this differentiation protocol, cells present morphological alterations, including long projections or cellular processes. 217,218 Other agents are described to induce differentiation of this cell line. One example is the protein kinase A activator forskolin, which, by down-regulating cyclin D1 expression, represses cell growth, via cell cycle arrest in the G 0 /G 1 phase, and induces cell differentiation characteristic with elongated processes and restoration of GFAP expression.…”

Section: C6 Cells: a Rat Glioma Cell Linementioning

confidence: 99%

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In vitro models for neurotoxicology research

et al. 2015

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Section: C6 Cells: a Rat Glioma Cell Linementioning

confidence: 99%

Section: C6 Cells: a Rat Glioma Cell Linementioning

confidence: 99%

In vitro models for neurotoxicology research

et al. 2015

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“…The rat C6 glioblastomas are an excellent cell line for modeling glial pathways due to their expression of glial proteins and their ability to differentiate into an astrocyte-like phenotype ( Ozawa et al, 2013 ; Chao et al, 2015 ). To better understand the function of MMPs in glia, we expressed GFP-dMMP and GFP Control constructs in C6 cells using Fugene 6 transfection reagent.…”

Section: Resultsmentioning

confidence: 99%

Expression of Drosophila Matrix Metalloproteinases in Cultured Cell Lines Alters Neural and Glial Cell Morphology

Hearst

Bednářová

Draughn

et al. 2021

Front. Cell Dev. Biol.

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Matrix metalloproteinases (MMPs) are zinc- and calcium- dependent endopeptidases that play pivotal roles in many biological processes. The expression of several MMPs in the central nervous system (CNS) have been shown to change in response to injury and various neurological/neurodegenerative disorders. While extracellular MMPs degrade the extracellular matrix (ECM) and regulate cell surface receptor signaling, the intracellular functions of MMPs or their roles in CNS disorders is unclear. Around 23 different MMPs are found in the human genome with overlapping function, making analysis of the intracellular role of human MMPs a daunting task. However, the fruit fly Drosophila melanogaster genome encodes only two MMPs: dMMP1 and dMMP2. To better understand the intracellular role of MMPs in the CNS, we expressed Green Fluorescent Protein (GFP)- tagged dMMPs in SH-SY5Y neuroblastoma cells and C6 glioblastoma cell lines. Lipofection of GFP-dMMPs in SH-SY5Y cells enhanced nuclear rupture and reduced cell viability (coupled with increased apoptosis) as compared to GFP alone. In non-liposomal transfection experiments, dMMP1 localizes to both the cytoplasm and the nucleus whereas dMMP2 had predominantly cytoplasmic localization in both neural and glial cell lines. Cytoplasmic localization demonstrated co-localization of dMMPs with cytoskeleton proteins which suggests a possible role of dMMPs in cell morphology. This was further supported by transient dMMP expression experiments that showed that dMMPs significantly increased neurite formation and length in neuronal cell lines. Inhibition of endogenous MMPs decreased neurite formation, length and βIII Tubulin protein levels in differentiated SH-SY5Y cells. Further, transient expression experiments showed similar changes in glial cell morphology, wherein dMMP expression increased glial process formation and process length. Interestingly, C6 cells expressing dMMPs had a glia-like appearance, suggesting MMPs may be involved in intracellular glial differentiation. Inhibition or suppression of endogenous MMPs in C6 cells increased process formation, increased process length, modulated GFAP protein expression, and induced distinct glial-like phenotypes. Taken together, our results strongly support the intracellular role that dMMPs can play in apoptosis, cytoskeleton remodeling, and cell differentiation. Our studies further reinforce the use of Drosophila MMPs to dissect out the precise mechanisms whereby they exert their intracellular roles in CNS disorders.

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“…The level of ACh may be adequately controlled in order for the ACh to bind to appropriate AChRs for the induction of biological reactivity in glial cells. The induced GFAP expression in C6 cells was suppressed by an AChE inhibitor and siRNA for AChE [17,18], which suggests that AChE dysfunction would induce an accumulation of ACh around glial cells and that ACh is an endogenous agonist for AChRs to control the condition of the glial cells.…”

Section: Discussionmentioning

confidence: 96%

Immunohistochemical analysis for acetylcholinesterase and choline acetyltransferase in mouse cerebral cortex after traumatic brain injury

Horio

Ozawa

Kamiie

et al. 2020

The Journal of Veterinary Medical Science

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The regulation of glial cells, especially astrocytes and microglia, is important to prevent the exacerbation of a brain injury because over-reactive glial cells promote neuronal death. Acetylcholine (ACh), a neurotransmitter synthesized and hydrolyzed by choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), respectively, in the central nervous system, has the potential to regulate glial cells' states, i.e., non-reactive and reactive states. However, the expression levels of these ACh-related enzymes in areas containing reactive glial cells are unclear. Herein we immunohistochemically investigated the distributions of AChE and ChAT with reactive glial cells in the cryo-injured brain of mice as a traumatic brain injury model. Immunohistochemistry revealed AChE-and ChATimmunopositive signals in injured areas at 7 days post-injury. The signals were observed in and around GFAP-or CD68-immunopositive cells, and the numbers of cells doubly positive for GFAP/AChE, GFAP/ChAT, CD68/AChE, and CD68/ChAT were significantly increased in injured areas compared to sham-operated areas. Enzyme histochemistry for AChE showed intensely positive signals in injured areas. These results suggest that reactive astrocytes and microglia express and secrete AChE and ChAT in brain-injury areas. These glial cells may adjust the ACh concentration around themselves through the regulation of the expression of ACh-related enzymes in order to control their reactive states.

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Acetylcholine esterase is a regulator of GFAP expression and a target of dichlorvos in astrocytic differentiation of rat glioma C6 cells

Cited by 9 publications

References 23 publications

In vitro models for neurotoxicology research

In vitro models for neurotoxicology research

Expression of Drosophila Matrix Metalloproteinases in Cultured Cell Lines Alters Neural and Glial Cell Morphology

Immunohistochemical analysis for acetylcholinesterase and choline acetyltransferase in mouse cerebral cortex after traumatic brain injury

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