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

Pickett, Melissa A.; Dush, Michael; Nascone-Yoder, Nanette

doi:10.1242/jcs.209031

Cited by 4 publications

(5 citation statements)

References 42 publications

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“…Clustered regulatory interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) has become a popular genome editing tool in Xenopus laevis (Aslan et al, 2017; Banach et al, 2017; Bharathan & Dickinson, 2019; Carr et al, 2021; Delay et al, 2018; Delay et al, 2019; Ezawa et al, 2021; Feehan et al, 2017; Jaffe et al, 2016; Kato et al, 2021; Kim et al, 2021; Ledford et al, 2017; Martin et al, 2020; Naert et al, 2020; Ochi et al, 2017; Pickett et al, 2017; Smith et al, 2020; Square et al, 2020; Tandon et al, 2017; Tsujioka et al, 2017; Viet et al, 2020; Wang et al, 2015; Wen et al, 2019; Wyatt et al, 2021). Importantly, most studies to date delivered the CRISPR reagents using a microinjection into fertilized eggs.…”

Section: Introductionmentioning

confidence: 99%

Optimization of CRISPR/Cas9‐mediated gene disruption in Xenopus laevis using a phenotypic image analysis technique

et al. 2022

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The CRISPR/Cas9 method has become popular for gene disruption experiments in Xenopus laevis. However, the experimental conditions that influence the efficiency of CRISPR/Cas9 remain unclear. To that end, we developed an image analysis technique for the semi‐quantitative evaluation of the pigment phenotype resulting from the disruption of tyrosinase genes in X. laevis using a CRISPR/Cas9 approach, and then examined the effects of varying five experimental parameters (timing of the CRISPR reagent injection into developing embryos; amount of Cas9 mRNA in the injection reagent; total injection volume per embryo; number of injection sites per embryo; and the culture temperature of the injected embryos) on the gene disruption efficiency. The results of this systematic analysis suggest that the highest possible efficiency of target gene disruption can be achieved by injecting a total of 20 nL of the CRISPR reagent containing 1500 pg of Cas9 mRNA or 4 ng of Cas9 protein into two separate locations (10 nL each) of one‐cell stage embryos cultured at 22°C. This study also highlights the importance of balancing the experimental parameters for increasing gene disruption efficiency and provides valuable insights into the optimal conditions for applying the CRISPR/Cas9 system to new experimental organisms.

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Section: Introductionmentioning

confidence: 99%

Optimization of CRISPR/Cas9‐mediated gene disruption in Xenopus laevis using a phenotypic image analysis technique

et al. 2022

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“…For example, whereas CoMO‐injected endoderm cells exhibit levels of membrane‐localized beta‐catenin (a key component of adherens junctions) comparable to adjacent uninjected cells, Vangl2 MO‐injected cells display decreased or absent beta‐catenin (compare Figure F and K). Indeed, in an ex vivo cell dissociation/reaggregation assay designed to evaluate calcium‐dependent (ie, adherens junction‐mediated) intercellular adhesion (see Experimental Procedures), we detected significantly less reaggregation ( P < 0.05; Figure R) of cells derived from Vangl2 MO‐injected gut tubes than from CoMO‐injected guts (compare Figures N,O and P,Q), suggesting morphant cells have fewer and/or less stable adherens junctions. Despite their decreased adhesion, Vangl2‐deficient cells remain viable, as we do not detect apoptotic markers in the Vangl2 MO‐injected population (not shown).…”

Section: Resultsmentioning

confidence: 95%

Vangl2 coordinates cell rearrangements during gut elongation

Dush

Nascone-Yoder

2019

Developmental Dynamics

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Background The embryonic gut tube undergoes extensive lengthening to generate the surface area required for nutrient absorption across the digestive epithelium. In Xenopus, narrowing and elongation of the tube is driven by radial rearrangements of its core of endoderm cells, a process that concomitantly opens the gut lumen and facilitates epithelial morphogenesis. How endoderm rearrangements are properly oriented and coordinated to achieve this complex morphogenetic outcome is unknown. Results We find that, prior to gut elongation, the core Wnt/PCP component Vangl2 becomes enriched at both the anterior and apical aspects of individual endoderm cells. In Vangl2‐depleted guts, the cells remain unpolarized, down‐regulate cell‐cell adhesion proteins, and, consequently, fail to rearrange, leading to a short gut with an occluded lumen and undifferentiated epithelium. In contrast, endoderm cells with ectopic Vangl2 protein acquire abnormal polarity and adhesive contacts. As a result, endoderm cells also fail to rearrange properly and undergo ectopic differentiation, resulting in guts with multiple torturous lumens, irregular epithelial architecture, and variable intestinal topologies. Conclusions Asymmetrical enrichment of Vangl2 in individual gut endoderm cells orients polarity and adhesion during radial rearrangements, coordinating digestive epithelial morphogenesis and lumen formation with gut tube elongation.

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“…Acetylcholinesterase (AChE) is expressed in both neuronal and non‐neuronal cell types (Pickett, Dush, & Nascone‐Yoder, 2017). Our immunocytochemistry revealed that AChE co‐localized with MBP, Tuj1, and GFAP, suggesting that AChE expression is not specific oligodendrocyte (Figure S3).…”

Section: Discussionmentioning

confidence: 99%

Donepezil‐induced oligodendrocyte differentiation is mediated through estrogen receptors

Imamura

Arai

Dateki

et al. 2019

Journal of Neurochemistry

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Loss of oligodendrocytes, the myelin-forming cells of the central nervous system, and subsequent failure of myelin development result in serious neurological disorders such as multiple sclerosis. Using primary mouse embryonic neural stem cells (NSCs), we previously demonstrated that donepezil, an acetylcholinesterase inhibitor developed for the treatment of Alzheimer's disease, stimulates the differentiation of NSCs into oligodendrocytes and neurons, albeit at the expense of astrogenesis. However, the precise mechanisms underlying donepezil-induced differentiation remain unclear. In this study, we aimed at elucidating the molecular pathways contributing to donepezil-induced differentiation of mouse-induced pluripotent stem cell-derived neural stem cells (miPSC-NSCs). We used cell-based reporter gene arrays to investigate effects of donepezil on differentiation of miPSC-NSCs. Subsequently, we assessed the molecular pathway underlying donepezil action on differentiation of miPSC-NSCs into mature oligodendrocytes. Donepezil increased the transcriptional activity of estrogen response element under differentiating conditions. Moreover, estrogen receptors α (ERα) and β (ERβ) were highly expressed in MBP-positive mature oligodendrocytes. The ER antagonist ICI 182,780 abrogated the number of MBP-positive oligodendrocytes induced by donepezil, but showed no effect on the differentiation of miPSC-NSCs into Tuj1-positive neurons and GFAP-positive astrocytes. Furthermore, the donepezilinduced generation of mature oligodendrocytes from miPSC-NSC was significantly attenuated by antagonists and siRNA targeting ERα and ERβ. In conclusion, we demonstrated, for the first time, that donepezil-induced oligodendrogenesis is mediated through both ER subtypes, ERα and ERβ.

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

Cited by 4 publications

References 42 publications

Optimization of CRISPR/Cas9‐mediated gene disruption in Xenopus laevis using a phenotypic image analysis technique

Optimization of CRISPR/Cas9‐mediated gene disruption in Xenopus laevis using a phenotypic image analysis technique

Vangl2 coordinates cell rearrangements during gut elongation

Donepezil‐induced oligodendrocyte differentiation is mediated through estrogen receptors

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