Novel Assay Utilizing Fluorochrome-Tagged Physostigmine (Ph-F) to In Situ Detect Active Acetylcholinesterase (AChE) Induced during Apoptosis

Huang, Xuan; Lee, Brian; Johnson, Gary L.; Naleway, John J.; Guzikowski, Anthony P.; Dai, Wei; Darżynkiewicz, Zbigniew

doi:10.4161/cc.4.1.1322

Cited by 22 publications

(19 citation statements)

References 28 publications

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“…Furthermore, AChE activity has been shown in A549 cell lysates using Ellman assay. The presented strong correlation between apoptosis and AChE activity are in line with observations of previous investigators (Zhang et al, 2002;Huang et al, 2005). Our data support the hypothesis that AChE activity in SM treated A549 cells may be used as a marker of apoptotic cell death.…”

Section: Discussionsupporting

confidence: 94%

“…Though, it has been proposed that AChE may be an important component of the apoptotic pathway. However, the physiological relevance of these findings is uncertain as AChE inhibition does not prevent apoptotic cell death in all investigated cell types (Huang et al, 2005). The aim of our study was to investigate, whether SM can induce apoptotic cell death in lung cancer derived A549 cells and whether AChE activity is simultaneously increased.…”

Section: Introductionmentioning

confidence: 98%

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Apoptosis in sulfur mustard treated A549 cell cultures

et al. 2007

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Section: Discussionsupporting

confidence: 94%

Section: Introductionmentioning

confidence: 98%

Apoptosis in sulfur mustard treated A549 cell cultures

et al. 2007

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“…AChE has been proposed to play a pivotal role in apoptosome formation [5,6], and synaptic AChE with an extended N-terminus induces apoptosis in neurons by interacting with other proteins [7,8]. Moreover, the appearance of this enzyme is considered as a marker for apoptosis under certain circumstances [9][10][11]. However, it is unknown whether AChE is induced in apoptotic cells in vivo.…”

Section: Introductionmentioning

confidence: 99%

Acetylcholinesterase is associated with apoptosis in β cells and contributes to insulin-dependent diabetes mellitus pathogenesis

Bao¹,

Yang²,

Yu³

et al. 2012

ABBS

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Acetylcholinesterase (AChE) expression is pivotal during apoptosis. Indeed, AChE inhibitors partially protect cells from apoptosis. Insulin-dependent diabetes mellitus (IDDM) is characterized in part by pancreatic b-cell apoptosis. Here, we investigated the role of AChE in the development of IDDM and analyzed protective effects of AChE inhibitors. Multiple low-dose streptozotocin (MLD-STZ) administration resulted in IDDM in a mouse model. Western blot analysis, cytochemical staining, and immunofluorescence staining were used to detect AChE expression in MIN6 cells, primary b cells, and apoptotic pancreatic b cells of MLD-STZ-treated mice. AChE inhibitors were administered intraperitoneally to the MLD-STZ mice for 30 days. Blood glucose, plasma insulin, and creatine levels were measured, and glucose tolerance tests were performed. The effects of AChE inhibitors on MIN6 cells were also evaluated. AChE expression was induced in the apoptotic MIN6 cells and primary b cells in vitro and pancreatic islets in vivo when treated with STZ. Induction and progressive accumulation of AChE in the pancreatic islets were associated with apoptotic b cells during IDDM development. The administration of AChE inhibitors effectively decreased hyperglycemia and incidence of diabetes, and restored plasma insulin levels and plasma creatine clearance in the MLD-STZ mice. AChE inhibitors partially protected MIN6 cells from the damage caused by STZ treatment. Induction and accumulation of AChE in pancreatic islets and the protective effects of AChE inhibitors on the onset and development of IDDM indicate a close relationship between AChE and IDDM.

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“…So far, assays for caspases have received considerable attention for apoptosis imaging;18, 57 however, developing cholinesterase assay principles for apoptosis imaging is still on demand. Only a few examples were reported58, 59 but suffered from puzzles of background fluorescence and washing difficulty. To our gratifying surprise, the present programmable self‐assembly is envisaged to be a smart strategy for imaging of live cell apoptosis with high fluorescence contrast.…”

Section: Resultsmentioning

confidence: 99%

Sequentially Programmable and Cellularly Selective Assembly of Fluorescent Polymerized Vesicles for Monitoring Cell Apoptosis

et al. 2017

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The introduction of controlled self‐assembly into living organisms opens up desired biomedical applications in wide areas including bioimaging/assays, drug delivery, and tissue engineering. Besides the enzyme‐activated examples reported before, controlled self‐assembly under integrated stimuli, especially in the form of sequential input, is unprecedented and ultimately challenging. This study reports a programmable self‐assembling strategy in living cells under sequentially integrated control of both endogenous and exogenous stimuli. Fluorescent polymerized vesicles are constructed by using cholinesterase conversion followed by photopolymerization and thermochromism. Furthermore, as a proof‐of‐principle application, the cell apoptosis involved in the overexpression of cholinesterase in virtue of the generated fluorescence is monitored, showing potential in screening apoptosis‐inducing drugs. The approach exhibits multiple advantages for bioimaging in living cells, including specificity to cholinesterase, red emission, wash free, high signal‐to‐noise ratio.

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Novel Assay Utilizing Fluorochrome-Tagged Physostigmine (Ph-F) to In Situ Detect Active Acetylcholinesterase (AChE) Induced during Apoptosis

Cited by 22 publications

References 28 publications

Apoptosis in sulfur mustard treated A549 cell cultures

Apoptosis in sulfur mustard treated A549 cell cultures

Acetylcholinesterase is associated with apoptosis in β cells and contributes to insulin-dependent diabetes mellitus pathogenesis

Sequentially Programmable and Cellularly Selective Assembly of Fluorescent Polymerized Vesicles for Monitoring Cell Apoptosis

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Novel Assay Utilizing Fluorochrome-Tagged Physostigmine (Ph-F) to In Situ Detect Active Acetylcholinesterase (AChE) Induced during Apoptosis

Cited by 22 publications

References 28 publications

Apoptosis in sulfur mustard treated A549 cell cultures

Apoptosis in sulfur mustard treated A549 cell cultures

Acetylcholinesterase is associated with apoptosis in &beta; cells and contributes to insulin-dependent diabetes mellitus pathogenesis

Sequentially Programmable and Cellularly Selective Assembly of Fluorescent Polymerized Vesicles for Monitoring Cell Apoptosis

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Acetylcholinesterase is associated with apoptosis in β cells and contributes to insulin-dependent diabetes mellitus pathogenesis