Characterization of acetylcholinesterase expression and secretion during osteoblast differentiation

Inkson, Colette A.; Brabbs, Alex C.; Grewal, T. S.; Skerry, Timothy M.; Genever, Paul G.

doi:10.1016/j.bone.2004.05.026

Cited by 68 publications

(53 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Due to amino acid sequence homology of AChE with cell adhesion proteins, such as gliotactin, glutactin, and neuroligins, AChE has been shown capable of promoting cellular adhesion of neurons and osteoblasts [Inkson et al, 2004;Johnson and Moore, 2004]. AChE may promote adhesion of epithelial cell lines, such as HeLa and LLC-MK2 cells, however, this study showed AChE is localized to the perinuclear region of these cells, and not on the plasma membrane, indicating involvement of other cellular factors in adhesion of these types of cell.…”

Section: Discussionmentioning

confidence: 99%

“…Forty micrograms of crude protein lysates were electrophoresed on a 10% SDS-PAGE gel for immunoblot analysis. Primary anti-huAChE monoclonal antibody (Clone 46, BD Biosciences, Mississauga, ON) has previously been used to detect AChE in immunoblot, immunohistochemical, and immunocytochemical assays [Boudreau-Lariviere and Jasmin, 1999;Meshorer et al, 2002;Inkson et al, 2004]. Anti-huAChE was diluted 1:1,500 in blocking buffer (5% non-fat dehydrated milk in PBS/0.3% Tween-20) and secondary anti-mouse IgG conjugated to HRP (Cell Signaling Technology, Beverly, MA) was diluted 1:1,000 in blocking buffer.…”

Section: Immunoblot Analysis Of Achementioning

confidence: 99%

See 1 more Smart Citation

Differential localization of acetylcholinesterase in neuronal and non‐neuronal cells

Thullbery

Cox

Schule

et al. 2005

J of Cellular Biochemistry

View full text Add to dashboard Cite

Acetylcholinesterase (AChE) expression is regulated in cell types at the transcriptional and translational levels. In this study, we characterized and compared AChE catalytic activity, mRNA, protein expression, and protein localization in a variety of neuronal (SH-SY5Y neuroblastoma and primary cerebellar granule neurons (CGN)) and non-neuronal (LLC-MK2, HeLa, THP-1, and primary astrocytes) cell types. All cell lines expressed AChE catalytic activity; however the levels of AChE-specific activity were higher in neuronal cells than in the non-neuronal cell types. CGN expressed significantly more AChE activity than SH-SY5Y cells. All cell lines analyzed expressed AChE protein at equivalent levels, as well as mRNA splice variants. Localization of AChE was characterized by immunofluorescence and confocal microscopy. SH-SY5Y, CGN, and nerve-growth factor-differentiated PC-12 cells exhibited a pattern of AChE localization characterized as diffuse in the cytoplasm and punctate staining along neurites and on the plasma membrane. The localization in HeLa, LLC-MK2, fibroblasts, and undifferentiated PC-12 cells was significantly different than in neuronal cells-AChE was intensely localized in the perinuclear region, without staining near or on the plasma membrane. Based on the evidence presented here, we hypothesize that the presence of AChE protein doesn't correlate with catalytic activity, and the diffuse cytoplasmic and plasma membrane localization of AChE is a property of neuronal cell types. Keywords acetylcholinesterase; confocal microscopy; cerebellar granule neuron; neuroblastoma; SH-SY5Y; HeLa; PC-12 Acetylcholinesterase (AChE) belongs to the serine hydrolase family that contains the α/β hydrolase fold as a common structural element [Ollis et al., 1992]. The catalytic active site of AChE is made up of a precise arrangement of active site functional groups that catalyze the hydrolytic breakdown of esters that bear quaternary ammonium groups. The biological importance of AChE-catalyzed hydrolysis is manifested in the rapid termination of the neuronal impulse that occurs when acetylcholine (ACh) is released into the synaptic cleft. AChE rapidly hydrolyzes ACh into acetic acid and choline at extremely fast turnover rates [Taylor and Radic, 1994].

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Immunoblot Analysis Of Achementioning

confidence: 99%

Differential localization of acetylcholinesterase in neuronal and non‐neuronal cells

Thullbery

Cox

Schule

et al. 2005

J of Cellular Biochemistry

View full text Add to dashboard Cite

show abstract

“…Inkson et al [183] reported about the production of AChE in osteoblasts which is believed to be involved in cell-matrix interaction in the bone. Our group was able to show that osteoblasts express all necessary components for the synthesis and release of ACh [184].…”

Section: Musculoskeletal Systemmentioning

confidence: 99%

The Non-Neuronal Cholinergic System in Health and Disease

Beckmann¹,

Lips²

2013

Pharmacology

102

View full text Add to dashboard Cite

Acetylcholine (ACh) is not only a neurotransmitter but is an ancient molecule that can be released by and act on non-neuronal cells. In these cells the system of ACh-synthesizing enzymes, transporters, receptors and degrading enzymes is termed the non-neuronal cholinergic system (NNCS). There is increasing evidence that the NNCS is dysregulated in various diseases and can have an influence on their pathology. However, for many organ systems not much is known about the expression and function of the NNCS. Thus, this review focusses on the role of the NNCS in different organ systems in health and disease. Dysregulation of ACh synthesis and release, mutations or polymorphisms in genes encoding NNCS components, and auto-antibodies against NNCS components are common factors influencing disease progression. Pharmacological agents targeting the NNCS are already successfully in clinical use for some disorders, indicating that interfering with this system is very promising and more research is needed to elucidate the role of the NNCS in different tissues and pathological states.

show abstract

“…This esterase activity is the target of widely used pesticides and pharmaceuticals (Mileson et al, 1998;Pope et al, 2005), yet the broad expression of AChE outside of the nervous system (Anderson et al, 2008;Bertrand et al, 2001;Bicker et al, 2004;Drews, 1975) and similarity to adhesion molecules (Botti et al, 1998;Darboux et al, 1996) suggest that it has additional functions. Indeed, in neuronal and non-neuronal cell lines, AChE promotes cell-substrate adhesion (Inkson et al, 2004;Johnson and Moore, 1999;Sharma et al, 2001;Syed et al, 2008), polarized cell migration (Anderson et al, 2008), cytoskeletal organization (Dupree and Bigbee, 1994;Keller et al, 2001) and cell differentiation (Grisaru et al, 1999;Xiang et al, 2008), independently of its esterase activity (Layer et al, 1993). However, the in vivo relevance of this putative multi-functionality is poorly substantiated, especially in non-neuronal contexts (Vogel-Hopker et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Acetylcholinesterase plays a non-neuronal, non-esterase role in organogenesis

2017

View full text Add to dashboard Cite

Acetylcholinesterase (AChE) is crucial for degrading acetylcholine at cholinergic synapses. In vitro studies suggest that, in addition to its role in nervous system signaling, AChE can also modulate nonneuronal cell properties, although it remains controversial whether AChE functions in this capacity in vivo. Here, we show that AChE plays an essential non-classical role in vertebrate gut morphogenesis. Exposure of Xenopus embryos to AChE-inhibiting chemicals results in severe defects in intestinal development. Tissuetargeted loss-of-function assays (via microinjection of antisense morpholino or CRISPR-Cas9) confirm that AChE is specifically required in the gut endoderm tissue, a non-neuronal cell population, where it mediates adhesion to fibronectin and regulates cell rearrangement events that drive gut lengthening and digestive epithelial morphogenesis. Notably, the classical esterase activity of AChE is dispensable for this activity. As AChE is deeply conserved, widely expressed outside of the nervous system, and the target of many environmental chemicals, these results have wide-reaching implications for development and toxicology.

show abstract

Characterization of acetylcholinesterase expression and secretion during osteoblast differentiation

Cited by 68 publications

References 48 publications

Differential localization of acetylcholinesterase in neuronal and non‐neuronal cells

Differential localization of acetylcholinesterase in neuronal and non‐neuronal cells

The Non-Neuronal Cholinergic System in Health and Disease

Acetylcholinesterase plays a non-neuronal, non-esterase role in organogenesis

Contact Info

Product

Resources

About