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Biology subjects

Tarnok, K.

Publications and source records attributed to Tarnok, K..

5 recordsLinked to original sources

Glia cells are selectively sensitive to nanosized titanium dioxide mineral forms

Nanosized titanium dioxide is widely used by the industry e.g. in pigments, suncreams and food colors. Its environmental and biological effects have been investigated in the past, however, few studies have focused on its crystal structure-specific effects. In our experiments, the toxicity of two types of nanoparticles was examined on primary neural cultures with different cell-compositions, using MTT and LDH assays. Primary murine cell cultures containing only astroglia cells originated from two brain regions, as well as mixed neurons and glia cells or microglia cells exclusively, were treated with anatase and rutile TiO2 nanoparticles at varying concentrations for 24 or 48 hours. Our results show that neither anatase nor rutile nanoparticles reduced viability in cell cultures containing a mixture of neurons and glial cells, independently of the applied concentration and treatment time. Rutile but not anatase form induced cell death in cortical astroglia cultures already at 24 hours of treatment above 10 {micro}g/mL, while hippocampus-derived glial cultures were much less sensitive to rutile. The rutile form also damaged microglia. These findings suggest that products containing rutile-form nano-titanium particles may pose a targeted risk to astroglia and microglial cells in the central nervous system.

neuroscience↗

Cell-type specific effects of Fusarium mycotoxins on primary neuronal and astroglial cells

Fumonisin B1, deoxynivalenol (DON) and zearalenone (ZEA) are toxic secondary metabolites produced by Fusarium molds. These mycotoxins are common food and feed pollutants and represent a risk for human and animal health. Although the mycotoxins produced by this genus can cross the blood-brain-barrier (BBB) in many species, their effect on neuronal function remains unclear. We investigated cell viability effects of these toxins on specified neural cell types, including mouse primary neuronal, astroglial and mixed cell cultures 24 or 48 hours after mycotoxin administration. Cell viability assay revealed that DON decreased cell viability in a dose-dependent manner, independently from the cultures type. Fumonisin B1 increased cell viability significantly on astroglial and mixed cell cultures in lower doses, while it exerted a highly toxic effect in 50 {micro}M. ZEA had significant effects on all culture type in 10 nM by increasing the cell viability. Since ZEA is a mycoestrogen, we analyzed the effects of ZEA on the expression of estrogen receptor isotypes ER and ER{beta} and mitochondrial voltage-dependent anion channel (VDAC1) by qRT-PCR. In neuronal and mixed cultures, ZEA administration decreased ER expression, while in astroglial cultures, it induced the opposite effect. ER{beta} and VDAC1 expression was not altered by ZEA in either culture types. ZEA also affected the firing pattern of neurons by enhancing the burst frequency. Our results demonstrate that Fusarium mycotoxins are acting on a cell specific manner in the brain tissue.

neuroscience↗

Multifactorial approach is needed to unravel the maturation phases of human neurons derived from induced pluripotent stem cells

Neurons derived from induced pluripotent stem cells (h-iPSC-Ns) provide an invaluable model for studying the physiological aspects of neuronal development and diseases. However, multiple studies have also demonstrated that h-iPSC-Ns exhibit a high degree of functional and epigenetic diversity. Due to the imprecise characterization and significant variation among the currently available maturation protocols, it is essential to establish a set of criteria to standardize models and accurately characterize and define the developmental properties of neurons derived from iPSCs. In this study, we conducted a comprehensive analysis of the h-iPSC-Ns via electrophysiological and microscopic techniques to follow their functional development at the cellular and network levels. This enabled us to provide a thorough description of the maturation process of h-iPSC-Ns over a 10-week period in vitro. Specifically, we have used conventional whole-cell patch-clamp and dynamic clamp techniques, alongside morphometry, to assess the characteristics of maturing h-iPSC-Ns. Additionally, we utilized calcium imaging to monitor the progression of synaptic activity and network communication. At the single cell level, human neurons exhibited gradually decreasing membrane resistance in parallel with improved excitability by 5 weeks of maturation. Their firing profiles were consistent with those of mature regular firing type of neurons. At the network level we observed the development of abundant fast glutamatergic and depolarizing GABAergic synaptic connections together with synchronized network activity. The identified sequence of differentiation events are consistent and offers a robust framework for developing targeted experiments at varying stages of neuronal maturation. This framework allows for the use of different, age-related methodologies or a singular set of experiments for a cultures maturation.

neuroscience↗

Redox-dependent condensation and cytoplasmic granulation by human ssDNA binding protein 1 delineate roles in oxidative stress response

Human single-stranded DNA binding protein 1 (hSSB1/NABP2/OBFC2B) plays central roles in the repair of DNA breaks and oxidized DNA lesions. Here we show that hSSB1 undergoes liquid-liquid phase separation (LLPS) that is redox-dependent and requires the presence of single-stranded DNA or RNA, features that are distinct from those of LLPS by bacterial SSB. hSSB1 nucleoprotein droplets form under physiological ionic conditions, in response to treatment resulting in cellular oxidative stress. hSSB1s intrinsically disordered region (IDR) is indispensable for LLPS, whereas all three cysteine residues of the oligonucleotide/oligosaccharide-binding (OB) fold are necessary to maintain redox-sensitive droplet formation. Proteins interacting with hSSB1 show selective enrichment inside hSSB1 droplets, suggesting tight content control and recruitment functions for the condensates. While these features appear instrumental for genome repair, we also detected hSSB1 condensates in the cytoplasm in response to oxidative stress in various cell lines. hSSB1 condensates colocalize with stress granules, implying unexplored extranuclear roles in cellular stress response. Our results suggest novel, condensation-linked roles for hSSB1, linking genome repair and cytoplasmic defense.

biochemistry↗

Phase separation by ssDNA binding protein controlled via protein-protein and protein-DNA interactions

Bacterial single stranded (ss) DNA-binding proteins (SSB) are essential for the replication and maintenance of the genome. SSBs share a conserved ssDNA-binding domain, a less conserved intrinsically disordered linker (IDL) and a highly conserved C-terminal peptide (CTP) motif that mediates a wide array of protein-protein interactions with DNA-metabolizing proteins. Here we show that the E. coli SSB protein forms liquid-liquid phase separated condensates in cellular-like conditions through multifaceted interactions involving all structural regions of the protein. SSB, ssDNA and SSB-interacting molecules are highly concentrated within the condensates, whereas phase separation is overall regulated by the stoichiometry of SSB and ssDNA. Together with recent results on subcellular SSB localization patterns, our results point to a conserved mechanism by which bacterial cells store a pool of SSB and SSB-interacting proteins. Dynamic phase separation enables rapid mobilization of this protein pool to protect exposed ssDNA and repair genomic loci affected by DNA damage.

biochemistry↗