Team 1

Scientific profile

About department
Employees

Research Profile of the Team

The Team conducts innovative research on the molecular mechanisms underlying neuroprotection and neurotoxicity. The research encompasses three thematically interconnected areas, integrating modern approaches at the interface of neurobiology, molecular biology, and toxicology. The Team’s objectives include not only advancing the understanding of the pathomechanisms of central nervous system (CNS) injury but also identifying potential therapeutic targets and risk factors relevant from a translational medicine perspective.

The first research area focuses on developing novel therapeutic strategies for acute brain injuries, such as ischemic stroke and perinatal asphyxia. This approach is based on the selective modulation of membrane-bound estrogen receptors to activate neuroprotective mechanisms.

The second line of research centers on evaluating the impact of semaglutide—a glucagon-like peptide-1 receptor (GLP-1R) agonist used in the treatment of type 2 diabetes and obesity – on prenatal CNS development. This project aims to elucidate how activation of the GLP-1 signaling pathway during gestation influences neurogenesis, synaptogenesis, and cognitive functions in the offspring, thereby identifying potential risks associated with early developmental exposure to this pharmaceutical agent.

The third research focus addresses the neurotoxic effects of xenobiotics, with particular emphasis on benzophenone-3 (BP-3), a widely used chemical UV filter present in cosmetics and various consumer products. The Team investigates the neurotoxic impact of BP-3 on the developing CNS, placing special emphasis on the consequences of prenatal exposure. These studies encompass not only directly exposed individuals (F0 generation) but also their progeny (F1) and subsequent generations (F2), allowing differentiation between multigenerational effects – arising from direct exposure of successive generations and transgenerational effects, defined as heritable phenotypic changes observed in descendants lacking direct contact with the toxicant. Particular attention is given to epigenetic and molecular mechanisms potentially underlying persistent modifications of neurobiological functions, which may have significant implications for population health.

The Team integrates advanced molecular biology, developmental neurobiology, and environmental toxicology techniques to comprehensively investigate the mechanisms underpinning neuroprotection and neurotoxicity. This multidisciplinary approach facilitates the identification of novel, precise therapeutic targets and enhances understanding of the molecular and epigenetic bases of neurotoxic risk. The conducted research not only deepens insight into CNS injury pathogenesis but also contributes to the development of innovative therapeutic and preventive strategies. The findings hold strong translational potential, supporting the creation of more effective treatments and reducing the adverse impact of environmental factors on brain health.

Moreover, the Team’s work provides critical evidence for health policy and public health initiatives aimed at protecting vulnerable populations, including pregnant women and developing fetuses. Our research has been frequently cited by prominent international organizations and expert bodies, including the European Union’s Human Biomonitoring for Europe program, the U.S. Food and Drug Administration (FDA), the Expert Panel for Cosmetic Ingredient Safety, the Organization for Economic Co-operation and Development (OECD), the European Chemicals Agency (ECHA), the French Agency for Food, Environmental and Occupational Health & Safety (ANSES), and the California Environmental Protection Agency (CalEPA).

Research Models

Mouse model of photothrombotic stroke
Prenatal exposure models for neurotoxic substances, including intragenerational and transgenerational effect paradigms
Cellular models of stroke and Alzheimer’s disease
Primary cultures of mouse neural cells
Cultures of human microglial cells
Cultures of human endothelial cells

Applied Research Techniques

Molecular Analyses:

Quantitative analysis of gene expression using real-time PCR (qPCR).
Comprehensive transcriptomic profiling utilizing microarray technology.
MicroRNA (miRNA) expression profiling for the identification of non-coding RNA signatures associated with disease pathogenesis.
Evaluation of protein levels and post-translational modifications using immunochemical techniques, including Western blotting and enzyme-linked immunosorbent assay (ELISA).

Epigenetic Analyses:

Assessment of global and gene-specific DNA methylation, including promoter methylation status.
Measurement of enzymatic activity of chromatin-modifying enzymes, such as DNA methyltransferases (DNMTs), histone acetyltransferases (HATs), histone deacetylases (HDACs), and sirtuins.
Analysis of RNA methylation (m6A) modifications and their regulatory impact on gene expression.
Investigation of SUMOylation as a post-translational protein modification affecting chromatin dynamics and cellular stress responses.
Enrichment analyses to identify disrupted molecular pathways and transcriptional regulatory networks.
Functional modulation of gene expression through RNA interference (RNAi) using small interfering RNAs (siRNAs) to silence target genes involved in key neurodegenerative mechanisms.

Blood–Brain Barrier (BBB) Integrity and Permeability Assessment:

In vitro evaluation using a tri-cellular co-culture model comprising human brain microvascular endothelial cells, astrocytes, and neurons.
Measurement of paracellular permeability using model compounds and calculation of the apparent permeability coefficient (Papp).

Biochemical Analyses:

Cell Death Mechanisms:

Apoptosis:

- Determination of caspase-3, -8, and -9 activity using commercially available fluorometric and luminescent assay kits (e.g., Caspase-Glo®, Apo-ONE®).
- Detection of DNA fragmentation by TUNEL assay.
- Quantification of phosphatidylserine externalization using Annexin V conjugates in flow cytometry.
- Measurement of mitochondrial membrane potential (ΔΨm) using fluorogenic dyes such as JC-1 or TMRE.
- Morphological assessment of apoptotic features (chromatin condensation, nuclear fragmentation, cytoplasmic blebbing) using fluorescence microscopy (e.g., Hoechst 33342).
- Gene and protein expression analysis of key pro-apoptotic and anti-apoptotic regulators (e.g., BAX, BCL-2, p53, cytochrome c).

Autophagy:

- Use of commercial kits (e.g., Autophagy Assay Kit, CYTO-ID® Autophagy Detection Kit) to quantify autophagic flux and visualize autophagosomes in live cells.
- Expression analysis of autophagy-related genes using qPCR, Western blot, and immunocytochemistry.

Oxidative Stress:

- Quantitative determination of reactive oxygen species (ROS), enzymatic activity of antioxidants (e.g., SOD, catalase, glutathione peroxidase), and levels of reduced and oxidized glutathione (GSH/GSSG).
- Analysis of oxidative stress-related genes and proteins by qPCR and Western blot.

Ferroptosis:

- Assessment of GPX4 activity, intracellular ferrous iron (Fe²⁺) content, lipid peroxidation, and expression of ferroptosis-regulating genes.

Necrosis and Necroptosis:

- Evaluation of lactate dehydrogenase (LDH) release as a marker of membrane disruption.
- Analysis of RIPK1 and RIPK3 gen and protein levels, key mediators of programmed necrosis (necroptosis).