Dataset Information

The assessment of hiPSC-derived Neural Stem Cells after a repeated-dose bisphenol A exposure

ABSTRACT: The present study evaluated the effects of a repeated-dose exposure of environmentally relevant BPA concentrations during the in vitro 3D neural induction of human induced pluripotent stem cells (hiPSCs), emulating a chronic exposure scenario. Quantitative proteomics at D21 revealed several differentially abundant proteins across all BPA-treated groups with important functions in NSC proliferation and maintenance.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Bachuki Shashikadze

LAB HEAD: Thomas Fröhlich

PROVIDER: PXD042045 | Pride | 2023-09-06

REPOSITORIES: Pride

ACCESS DATA

Publications

Proteomics and disease network associations evaluation of environmentally relevant Bisphenol A concentrations in a human 3D neural stem cell model.

Horánszky Alex A Shashikadze Bachuki B Elkhateib Radwa R Lombardo Salvo Danilo SD Lamberto Federica F Zana Melinda M Menche Jörg J Fröhlich Thomas T Dinnyés András A

Frontiers in cell and developmental biology 20230816

Bisphenol A (BPA) exposure is associated with a plethora of neurodevelopmental abnormalities and brain disorders. Previous studies have demonstrated BPA-induced perturbations to critical neural stem cell (NSC) characteristics, such as proliferation and differentiation, although the underlying molecular mechanisms remain under debate. The present study evaluated the effects of a repeated-dose exposure of environmentally relevant BPA concentrations during the <i>in vitro</i> 3D neural induction of ...[more]

PMID: 37664457

Similar Datasets

Project description:Formaldehyde fixation is widely used for long-term maintenance of tissue. However, due to formaldehyde-induced cross-links, fixated tissue proteins are difficult to extract hampering the performance of mass spectrometry (MS)-proteomic analysis on formaldehyde-fixated tissue. Recent years saw the use of different combinations of high-temperature and solubilizing agents (usually derived from antigen retrieval techniques) to unravel formaldehyde-fixated paraffin-embedded tissue proteomes. However, in order to achieve protein extraction yields similar to fresh-frozen tissue high-temperature heating is necessary. Such harsh extraction conditions may affect, labile post-translational modifications such as phosphorylations resulting in the loss of important protein information. The objective of the present work is to assess cleavable fixative reagents that allow tissue preservation as well as efficient protein extraction from fixated tissue for MS-proteomics under mild conditions. To this regard, we investigated disuccinimidyl tartrate (DST) and dithiobis[succinimidylpropionate] (DSP) as cleavable fixating reagents. These compounds crosslink proteins by reacting with amino groups leading to amide bond formation. Linkers can be cleaved with sodium metaperiodate (cis-diols) or reducing agents (disulfide bonds), respectively. Our results show that reversible protein crosslinking allows tissue fixation with morphology preservation comparable to formalin. In addition, cleavage of DSP improves protein recovery from fixated tissue by a factor of 18 and increases the number of identified proteins by approximately 20% under mild extraction conditions avoiding the need for sample boiling, which could affect labile post-translational modifications. A major advantage of DSP over formaldehyde is the introduction of well-defined protein modifications that can be taken into account during database searching. In contrast to DSP fixation, DST fixation followed by periodic cleavage, while effective, resulted in side reactions that prevented effective protein extraction and subsequent protein identification. Protein crosslinkers, which can be cleaved under mild conditions, are thus viable alternatives to formaldehyde as tissue fixatives facilitating protein analysis from paraffin embedded, fixated tissue.

Dataset Information

The assessment of hiPSC-derived Neural Stem Cells after a repeated-dose bisphenol A exposure

Publications

Proteomics and disease network associations evaluation of environmentally relevant Bisphenol A concentrations in a human 3D neural stem cell model.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure