The department has a total of ~140 m2, including laboratory, office, and storage space. Our culture room contains a laminar box, four CO2 incubators, a dissecting magnifying glass, and a microscope; we also have equipment for molecular biology (UV DNA/RNA decontamination box, incubators, shakers, PCR cyclers, centrifuges), biochemistry (homogenizers, sonicators, gel electrophoresis, Western blotting apparatus) and electrophysiology (micropipette puller, three fully equipped electrophysiology apparatus with rapid application systems, one newly built electrophysiology apparatus for recording from acute brain tissue sections). Our experimental research is conducted using a combination of advanced molecular biology methods (preparation of DNA expression vectors, targeted mutagenesis, lentiviral particle production, genotyping of mouse models), microscopy (classical confocal microscopy, live quantum dot microscopy, uPAINT, dSTORM), biochemistry (SDS-PAGE, "Western blotting", colorimetric essays) and electrophysiology (patch-clamp method in the whole-cell or single-channel configuration in cell cultures or brain sections). We maintain breeding colonies of mice carrying genes for NMDA receptor subunits with conditional "knock-out" (cKO-Grin2A, cKO-Grin2B, cKO-Grin2A/Grin2B, cKO-Grin1) or "knock-in" (globally expressing the Grin2aN615S gene; a model of sound-induced epileptic seizures).
The Department of Neurochemistry was established in 2018 to conduct cutting-edge research in regulating N-methyl-D-aspartate (NMDA) receptors in mammalian neurons under normal and pathological conditions. Specifically, we study the molecular mechanisms that regulate i) the surface numbers of NMDA receptors, including their maturation in the endoplasmic reticulum and transport across the Golgi apparatus, and ii) the surface mobility and localization of NMDA receptors in synaptic and extrasynaptic regions of mammalian neurons. We are also studying (iii) the functional impact of selected pathogenic variants in genes encoding GluN subunits of NMDA receptors associated with developing epilepsy and other neurological diseases and (iv) developing pharmacological modulators of NMDA receptors with unique mechanisms of action. Our results contribute to the understanding and future therapy of disorders associated with abnormal regulation of NMDA receptors, including, for example, epilepsy and Alzheimer's disease.
- The Institute of Organic Chemistry and Biochemistry CAS, Prague, Czech Republic (Mgr. et Mgr. Evžen Bouřa, Ph.D.)
- Institute of Physiology CAS, Prague, Czech Republic (RNDr. Martin Zápotocký, Ph.D.)
- National Institute of Mental Health, Klecany, Czech Republic (RNDr. Karel Valeš, Ph.D.)
- Biomedical Research Center, Hradec Králové, Czech Republic (doc. PharmDr. Ondřej Soukup, Ph.D. a PharmDr. Jan Korábečný, Ph.D.)
- Neuroscience Research Institute, Seoul National University College of Medicine, Soul, Republic of Korea (Assoc. Prof. Young Ho Suh, Ph.D.)