Modulation of the Distribution of Acetylcholinesterase Molecular Forms in a Murine Neuroblastoma × Sympathetic Ganglion Cell Hybrid Cell Line

Lazar, Monique; Vigny, Marc

doi:10.1111/j.1471-4159.1980.tb07860.x

Cited by 88 publications

(32 citation statements)

References 39 publications

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“…Previous work on this cell line (Johnson and Moore, 2000) has shown that there are no high salt-soluble isoforms present. These results on the distribution of AChE isoforms confirm others on neuroblastoma cell lines (Kimhi et al, 1980;Lazar and Vigny, 1980;Melone et al, 1987;Stieger et al, 1989). The increase in AChE levels seen on differentiation is well documented, and is frequently used as a differentiation marker in many cell types.…”

Section: Discussionsupporting

confidence: 88%

Association of the HNK‐1 epitope with the detergent‐soluble G4 isoform of acetylcholinesterase from human neuroblastoma cells

Johnson

Moore

2001

Intl J of Devlp Neuroscience

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The HNK-1 carbohydrate epitope is expressed in neural and natural killer cells and is a mediator of cell adhesion. It is well documented that acetylcholinesterase has a secondary function in cell adhesion and differentiation. The presence of HNK-1 on isoforms of Torpedo and Electrophorus acetylcholinesterase, as well as isoforms from the bovine central nervous system has been described. In this paper, we have investigated the association of the epitope with acetylcholinesterase from human neuroblastoma cells. Acetylcholinesterase was extracted, with or without detergent, purified on immunoaffinity columns and the isoforms separated by sucrose density gradient sedimentation. Secreted acetylcholinesterase, from spent serum-free culture medium, was similarly treated. The presence of the HNK-1 epitope was determined by ELISA using the anti-HNK-1 and Elec 39 monoclonal antibodies. The epitope was found to be associated with the detergent-soluble G4 isoform, but not with the hydrophilic G1 nor the secreted hydrophilic G4 isoforms. Likewise, no HNK-1 was observed associated with human erythrocyte acetylcholinesterase. These results indicate that acetylcholinesterase-G4, anchored in the extracellular membrane, is capable of mediating cell-substrate adhesion through HNK-1.

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Section: Discussionsupporting

confidence: 88%

Association of the HNK‐1 epitope with the detergent‐soluble G4 isoform of acetylcholinesterase from human neuroblastoma cells

Johnson

Moore

2001

Intl J of Devlp Neuroscience

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“…There is no chemical or structural basis for a difference in substrate affinity: butyrylcholine is simply a larger molecule that cannot access the more constrained active site of AChE. These results on neuroblastoma AChE and BChE levels, both cell-associated and in the medium, confirm those of other workers [45][46][47].…”

Section: Discussionsupporting

confidence: 79%

Cholinesterases modulate cell adhesion in human neuroblastoma cells in vitro

Johnson¹,

Moore²

2000

Intl J of Devlp Neuroscience

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Cholinesterases are expressed non-synaptically during embryonic development, neoplasia and neurodegeneration. We have investigated the effects of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and, conversely, anti-AChE and -BChE antibodies and inhibitors on cell adhesion and neurite outgrowth in human neuroblastoma cells. Analysis of cholinesterase levels and isoforms in undifferentiated and differentiated cells indicated a significant rise in AChE levels on differentiation. This increase was related to both cell-associated and secreted enzyme, and was predominantly the G4 isoform. BChE levels and isoforms, on the other hand, showed no significant variation. Coating the tissue culture plate with AChE stimulated neurite outgrowth, while BChE had an anti-adhesive effect. Cell adhesion was affected by the BChE inhibitor, ethopropazine, and the AChE peripheral site inhibitor, BW284c51, but not by eserine which binds to the active site. This indicates that the adhesion function is non-cholinergic, a finding supported by the lack of effect of AE-2, a monoclonal antibody that inhibits AChE, on cell adhesion. Four out of a panel of nine anti-AChE antibodies inhibited adhesion to varying degrees. Of these antibodies, two are catalytic, with epitopes associated with the peripheral anionic site of AChE, and the remaining two have epitopes overlapping this site. Neither of the two anti-BChE antibodies used had any effect on adhesion. These results indicate the importance of AChE in neuroblastoma cell adhesion and neurite outgrowth, and suggest that the peripheral anionic site may be involved in these processes.

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“…However, quantitative comparisons of ChE activity levels between NTera 2 cells and other tissue sources are difficult, since NTera 2 cells show varying levels of ChE when grown at different densities (data not shown). Such variation in ChE levels is also seen in cultured murine neuroblastoma cells and may reflect density-dependent differences in mitotic activity (Blume et al, 1973;Schneider, 1993;Vimard et al, 1993;Kimhi et al, 1980;Lazar and Vigny, 1980;Chang and Blume, 1976).…”

Section: Cholinesterase Is Expressed In Undifferentiated and Differenmentioning

confidence: 93%

Acetylcholinesterase expression in NTera 2 human neuronal cells: A model for developmental expression in the nervous system

Llanes

Collman

Hrin

et al. 1995

J of Neuroscience Research

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Acetylcholinesterase (AChE; EC 3.1.1.7) is expressed in the central nervous system in multiple molecular forms that may subserve multiple functions and may be selectively lost in neurodegenerative illnesses such as Alzheimer's disease. AChE expression has been studied in primary cultures of developing vertebrate nervous system, but investigation has been limited by the lack of a suitable human CNS surrogate cell model system for in vitro studies and the inability of primary brain cultures to provide large numbers of pure neurons. To develop an in vitro model for studies of neuronal AChE expression and function, we utilized a neuronally committed human teratocarcinoma cell line, NTera 2, that can be induced to differentiate to a post-mitotic CNS neuronal phenotype. We found that NTera 2 cells express multiple molecular forms of AChE that are similar to CNS-derived AChE isoforms in velocity sedimentation profile, anion exchange elution profile, and sensitivity to inhibitors. At least two forms of AChE are expressed (G1 and G4), similar to human and rodent brain, and induction of NTera 2 cell differentiation results in an increased G4/G1 ratio, which is characteristic of mature neurons. As in primary CNS neurons, AChE is present in NTera 2 cells in both the cytosolic fraction and in the outer membrane, and is also released in a soluble form. These observations indicate that NTera 2 cells provide a useful human model system for studies of expression of cell-associated and soluble cell-free AChE in developing and mature human neurons and for elucidating the potential role(s) of acetylcholinesterase metabolism in both normal development and neurodegenerative disease states.

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Modulation of the Distribution of Acetylcholinesterase Molecular Forms in a Murine Neuroblastoma × Sympathetic Ganglion Cell Hybrid Cell Line

Cited by 88 publications

References 39 publications

Association of the HNK‐1 epitope with the detergent‐soluble G4 isoform of acetylcholinesterase from human neuroblastoma cells

Association of the HNK‐1 epitope with the detergent‐soluble G4 isoform of acetylcholinesterase from human neuroblastoma cells

Cholinesterases modulate cell adhesion in human neuroblastoma cells in vitro

Acetylcholinesterase expression in NTera 2 human neuronal cells: A model for developmental expression in the nervous system

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