Laboratory of Neuropharmacology and Epigenetics

Laboratory of Neuropharmacology and Epigenetics

Historical view

The Laboratory of Neuropharmacology and Epigenetics was established on April 23, 2021 as part of the Department of Pharmacology at the Maj Institute of Pharmacology of the Polish Academy of Sciences.

The laboratory was established as a result of the transformation of the Laboratory of Molecular Neuroendocrinology, operating since August 7, 2018.

The laboratory is one of the five innovative research laboratories created as part of the project entitled "Modernization of the building and research infrastructure of the Institute of Pharmacology of the Polish Academy of Sciences in Krakow in order to create innovative laboratories for brain research" implemented in the years 2009-2013.

The funds for the establishment of the Laboratory came from the Innovative Economy Operational Program (2.1. Development of centers with high research potential, European Regional Development Fund, 2007-2013 perspective).

The idea of ​​establishing the Laboratory appeared in response to the competition entitled "Idea for a new laboratory" announced by the Director of the Institute of Pharmacology of the Polish Academy of Sciences, prof. Krzysztof Wędzony in 2009.

Research profile

Searching for substances with neuroprotective potential constituting the basis of new therapies for stroke, perinatal asphyxia and Alzheimer's disease.

Identifying molecular mechanisms of neurotoxicity caused by exposures of cells or animals to hormonally active substances present in the environment and their roles in etiology of nervous system diseases.

Tests and models

Primary cultures of rodent brain neurons carried out in dispersed and organotypic systems

Cultures of nerve stem cells and astrocytes

Cultures of human neurons derived from induced pluripotent stem cells (iPSCs)

Cellular and animal models of strokes, perinatal asphyxia and Alzheimer's disease

The most important discoveries

Selective modulation of the PPARγ receptor by amorfrutin B or the membrane fraction of estrogen receptors by PaPE-1 produces a neuroprotective effect in cellular models of cerebral hypoxia and ischemia. Moreover, PaPE-1 protects nerve cells from the toxic effects of amyloid-β

The aryl hydrocarbon receptor (AhR) can be a target for drugs to improve the pharmacotherapy of cerebral hypoxia, as demonstrated by both in vitro (neuronal cells exposed to hypoxia / ischemia) and in vivo (rats exposed perinatally to asphyxia) studies.

Prenatal exposure to the biocide - triclocarban impairs the signaling pathways of the ERα receptor (now ESR1), disrupts the expression of genes involved in neurogenesis and neurotransmission, and changes the epigenetic status in the brains of postnatal mice in a sex-specific manner.

Prenatal exposure to the DDT pesticide may cause depressive-like disorders, which is associated with a reduction in the levels of estrogen receptors ERα and GPR30 (now ESR1 and GPER1), hypermethylation of the genes encoding them, and global DNA hypomethylation in the mouse brain.

Exposures of embryonic neuronal cells to environmental pollutants, i.e., the pesticide DDT and its metabolite DDE, the chemical UV filter - benzophenone-3 or the biocide - triclocarban, disrupt the signaling pathways of estrogen receptors and xenobiotic receptors, and change the epigenetic status of mammalian brain cells.

Research methods

Assessment of cytotoxicity and cell survival, incl. measurement of lactate dehydrogenase (LDH) release, staining with calcein AM, NeuroFluor ™ NeuO.

Determination of markers of oxidative stress with the use of 2 ', 7'-dichlorodihydrofluorescein diacetate (H2DCFDA).

Study of the processes of apoptosis and autophagy by measuring the level of specific markers (including caspases, membrane mitochondrial potential, apoptotic bodies, autophagosomes).

 Molecular analyzes incl. measurement of gene expression by qPCR and microarrays, measurement of protein expression by ELISA and western blot methods, silencing of gene expression using specific siRNAs.

Epigenetic analyzes, incl. measurement of DNA methylation, both global and specific genes, measurement of the activity of enzymes involved in histone modification (HAT, HDAC, sirtuin), miRNA expression specific for central nervous system diseases.

Immunofluorescence detection of specific proteins using confocal microscopy.

Blood-brain barrier permeation testing using an artificial system containing nerve cells, astrocytes and endothelial cells.

Keywords

apoptosis, aryl hydrocarbon receptor (AhR), asphyxia, autophagy, bazedoxifene, benzophenone-3, 3,3'-diindolomethane (DIM), daidzein, DDE, DDT, DNA methylation, epigenetics, estrogen receptors (ER / ESR; GPR30 / GPER1), excitotoxicity, genistein, hypoxia, ischemia, microarrays, miRNA, molecular biology, neuroprotection, neurotoxicity, pesticides, peroxisome proliferator activated receptors (PPAR), phytoestrogens, primary neuronal cultures, raloxifene, retinoid X receptors (RXRs), selective estrogen receptor modulators (SERMs), selective aryl hydrocarbon receptor modulators (SAhRMs), siRNA, stroke, triclocarban, xenobiotic receptors