Identification of Three Small Molecules That Can Selectively Influence Cellular Manganese Levels in a Mouse Striatal Cell Model

Manganese (Mn) is a biologically essential metal, critical as a cofactor for numerous enzymes such a glutamine synthetase and kinases such as ataxia-telangiectasia mutated (ATM). Similar to other essential metals such as iron and zinc, proper levels of Mn need to be achieved while simultaneously being careful to avoid excess levels of Mn that can be neurotoxic. A lifetime of occupational exposure to Mn can often lead to a Parkinsonian condition, also known as “manganism”, characterized by impaired gait, muscle spasms, and tremors. Despite the importance of its regulation, the mechanisms underlying the transport and homeostasis of Mn are poorly understood. Rather than taking a protein or gene-targeted approach, our lab recently took a high-throughput-screening approach to identify 41 small molecules that could significantly increase or decrease intracellular Mn in a neuronal cell model. Here, we report characterization of these small molecules, which we refer to as the “Mn toolbox”. We adapted a Fura-2-based assay for measuring Mn concentration and for measuring relative concentrations of other divalent metals: nickel, copper, cobalt, and zinc. Of these 41 small molecules, we report here the identification of three that selectively influence cellular Mn but do not influence the other divalent metals tested. The patterns of activity across divalent metals and the discovery of Mn-selective small molecules has potential pharmacological and scientific utility.

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Characterization of electroporation for unbiased delivery of small molecules in high throughput screening applications (654.13)

The FASEB Journal ◽

10.1096/fasebj.28.1_supplement.654.13 ◽

2014 ◽

Vol 28 (S1) ◽

Author(s):

Johan Pihl ◽

Anna Karlsson ◽

Mattias Karlsson ◽

Susanne Lardell ◽

Paul Karila

Keyword(s):

Small Molecules ◽

High Throughput ◽

High Throughput Screening

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Improvement of mitochondrial function and dynamics by the metabolic enhancer piracetam in a human neuronal cell model of early Alzheimer's disease

Pharmacopsychiatry ◽

10.1055/s-0033-1353305 ◽

2013 ◽

Vol 46 (06) ◽

Author(s):

C Schiller ◽

D Miano ◽

K Leuner ◽

WE Müller

Keyword(s):

Alzheimer’S Disease ◽

Alzheimer's Disease ◽

Mitochondrial Function ◽

Cell Model ◽

Neuronal Cell ◽

Human Neuronal Cell ◽

Early Alzheimer’S Disease

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A Cellular Assay for the Identification and Characterization of Connexin Gap Junction Modulators

International Journal of Molecular Sciences ◽

10.3390/ijms22031417 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1417

Author(s):

Azeem Danish ◽

Robin Gedschold ◽

Sonja Hinz ◽

Anke C. Schiedel ◽

Dominik Thimm ◽

...

Keyword(s):

Gap Junctions ◽

Small Molecules ◽

High Throughput Screening ◽

Cell Communication ◽

Receptor Activation ◽

Adenosine A2a Receptor ◽

Intracellular Camp ◽

Donor Cells ◽

A2a Receptor ◽

Adenosine A2a

Connexin gap junctions (Cx GJs) enable the passage of small molecules and ions between cells and are therefore important for cell-to-cell communication. Their dysfunction is associated with diseases, and small molecules acting as modulators of GJs may therefore be useful as therapeutic drugs. To identify GJ modulators, suitable assays are needed that allow compound screening. In the present study, we established a novel assay utilizing HeLa cells recombinantly expressing Cx43. Donor cells additionally expressing the Gs protein-coupled adenosine A2A receptor, and biosensor cells expressing a cAMP-sensitive GloSensor luciferase were established. Adenosine A2A receptor activation in the donor cells using a selective agonist results in intracellular cAMP production. The negatively charged cAMP migrates via the Cx43 gap junctions to the biosensor cells and can there be measured by the cAMP-dependent luminescence signal. Cx43 GJ modulators can be expected to impact the transfer of cAMP from the donor to the biosensor cells, since cAMP transit is only possible via GJs. The new assay was validated by testing the standard GJ inhibitor carbenoxolon, which showed a concentration-dependent inhibition of the signal and an IC50 value that was consistent with previously reported values. The assay was demonstrated to be suitable for high-throughput screening.

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Biomedical application of photo-crosslinked gelatin hydrogels

Journal of Leather Science and Engineering ◽

10.1186/s42825-020-00043-y ◽

2021 ◽

Vol 3 (1) ◽

Author(s):

Lei Xiang ◽

Wenguo Cui

Keyword(s):

Drug Delivery ◽

Tissue Engineering ◽

Biomedical Application ◽

Biomedical Field ◽

Or Gene ◽

Tunable Mechanical Properties ◽

Bio Compatibility ◽

Regeneration Of Bone ◽

Development Prospects

Abstract During the past decades, photo-crosslinked gelatin hydrogel (methacrylated gelatin, GelMA) has gained a lot of attention due to its remarkable application in the biomedical field. It has been widely used in cell transplantation, cell culture and drug delivery, based on its crosslinking to form hydrogels with tunable mechanical properties and excellent bio-compatibility when exposed to light irradiation to mimic the micro-environment of native extracellular matrix (ECM). Because of its unique biofunctionality and mechanical tenability, it has also been widely applied in the repair and regeneration of bone, heart, cornea, epidermal tissue, cartilage, vascular, peripheral nerve, oral mucosa, and skeletal muscle et al. The purpose of this review is to summarize the recent application of GelMA in drug delivery and tissue engineering field. Moreover, this review article will briefly introduce both the development of GelMA and the characterization of GelMA. Finally, we discuss the challenges and future development prospects of GelMA as a tissue engineering material and drug or gene delivery carrier, hoping to contribute to accelerating the development of GelMA in the biomedical field. Graphical abstract

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High-Throughput Screening and Characterization of a High-Density Soybean Mutant Library Elucidate the Biosynthesis Pathway of Triterpenoid Saponins

Plant and Cell Physiology ◽

10.1093/pcp/pcz025 ◽

2019 ◽

Vol 60 (5) ◽

pp. 1082-1097 ◽

Cited By ~ 4

Author(s):

Panneerselvam Krishnamurthy ◽

Yukiko Fujisawa ◽

Yuya Takahashi ◽

Hanako Abe ◽

Kentaro Yamane ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

High Density ◽

Biosynthesis Pathway ◽

Mutant Library ◽

Triterpenoid Saponins

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Identification of Potential Key lncRNAs in the Context of Mouse Myeloid Differentiation by Systematic Transcriptomics Analysis

Genes ◽

10.3390/genes12050630 ◽

2021 ◽

Vol 12 (5) ◽

pp. 630

Author(s):

Yongqing Lan ◽

Meng Li ◽

Shuangli Mi

Keyword(s):

Transcription Factor ◽

Cell Model ◽

Myeloid Differentiation ◽

Rna Seq ◽

Hematopoietic Differentiation ◽

Granulocytic Differentiation ◽

Expression Of Genes ◽

Non Coding Rnas

Hematopoietic differentiation is a well-orchestrated process by many regulators such as transcription factor and long non-coding RNAs (lncRNAs). However, due to the large number of lncRNAs and the difficulty in determining their roles, the study of lncRNAs is a considerable challenge in hematopoietic differentiation. Here, through gene co-expression network analysis over RNA-seq data generated from representative types of mouse myeloid cells, we obtained a catalog of potential key lncRNAs in the context of mouse myeloid differentiation. Then, employing a widely used in vitro cell model, we screened a novel lncRNA, named Gdal1 (Granulocytic differentiation associated lncRNA 1), from this list and demonstrated that Gdal1 was required for granulocytic differentiation. Furthermore, knockdown of Cebpe, a principal transcription factor of granulocytic differentiation regulation, led to down-regulation of Gdal1, but not vice versa. In addition, expression of genes involved in myeloid differentiation and its regulation, such as Cebpa, were influenced in Gdal1 knockdown cells with differentiation blockage. We thus systematically identified myeloid differentiation associated lncRNAs and substantiated the identification by investigation of one of these lncRNAs on cellular phenotype and gene regulation levels. This study promotes our understanding of the regulation of myeloid differentiation and the characterization of roles of lncRNAs in hematopoietic system.

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The Novel Alpha-2 Adrenoceptor Inhibitor Beditin Reduces Cytotoxicity and Huntingtin Aggregates in Cell Models of Huntington’s Disease

Pharmaceuticals ◽

10.3390/ph14030257 ◽

2021 ◽

Vol 14 (3) ◽

pp. 257

Author(s):

Elisabeth Singer ◽

Lilit Hunanyan ◽

Magda M. Melkonyan ◽

Jonasz J. Weber ◽

Lusine Danielyan ◽

...

Keyword(s):

Huntington's Disease ◽

Huntington’S Disease ◽

Neurodegenerative Disorder ◽

Cell Model ◽

Resonance Energy Transfer ◽

Neuronal Cell ◽

Resonance Energy ◽

Terminal Deoxynucleotidyl Transferase ◽

Tunel Staining ◽

Cell Models

Huntington’s disease (HD) is a monogenetic neurodegenerative disorder characterized by the accumulation of polyglutamine-expanded huntingtin (mHTT). There is currently no cure, and therefore disease-slowing remedies are sought to alleviate symptoms of the multifaceted disorder. Encouraging findings in Alzheimer’s and Parkinson’s disease on alpha-2 adrenoceptor (α2-AR) inhibition have shown neuroprotective and aggregation-reducing effects in cell and animal models. Here, we analyzed the effect of beditin, a novel α2- adrenoceptor (AR) antagonist, on cell viability and mHTT protein levels in cell models of HD using Western blot, time-resolved Foerster resonance energy transfer (TR-FRET), lactate dehydrogenase (LDH) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) cytotoxicity assays. Beditin decreases cytotoxicity, as measured by TUNEL staining and LDH release, in a neuronal progenitor cell model (STHdh cells) of HD and decreases the aggregation propensity of HTT exon 1 fragments in an overexpression model using human embryonic kidney (HEK) 293T cells. α2-AR is a promising therapeutic target for further characterization in HD models. Our data allow us to suggest beditin as a valuable candidate for the pharmaceutical manipulation of α2-AR, as it is capable of modulating neuronal cell survival and the level of mHTT.

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