The Department of Neurochemistry studies molecular mechanisms that regulate the function of glutamate receptors in the mammalian central nervous system. We also aim to develop novel pharmacological compounds that act on glutamate receptors for the treatment of cognitive decline caused by neurodegeneration. Our experimental approach includes microscopy, biochemistry, and electrophysiology in cultured hippocampal neurones and human fibroblasts derived from patients with neurodegenerative diseases. Our long-term goal is to develop “personalised” compounds for treating specific neurodegenerative conditions including Alzheimer disease.
From the left: Emily Langore, Štěpán Kortus, Marek Ladislav, Martin Horák, Jakub Netolický, Marharyta Kolcheva, Kristýna Řeháková, Matěj Hotovec, Petra Baráčková, Anna Misiachna
Mgr. Petra Baráčková
Mgr. Martin Horák, Ph.D.
Ing. Štěpán Kortus, Ph.D.
Mgr. Marek Ladislav, Ph.D.
Mgr. Petra Baráčková
Mgr. Marharyta Kolcheva
Emily Langore, MSc.
Mgr. Anna Misiachna
Mgr. Jakub Netolický
Bc. Matěj Hotovec
The pathogenic mutation in the GluN1 subunit regulates NMDA receptors in mammalian neurons
Using in silico modeling, microscopy, biochemistry and electrophysiology in cultured HEK293 cells and hippocampal neurons, we found that the pathogenic mutation S688Y in the GluN1 subunit of the NMDA receptor (NMDAR) significantly increases affinity for glycine and also decreases surface expression of NMDARs containing the GluN3A subunit. We further found that the S688Y mutation reduces both Ca2+ influx through NMDAR and NMDA-induced excitotoxicity. See more in the article Skrenkova et al. 2020.
1) A glycine binding site in the GluN1 subunit with the amino acid residue S688 (shown in red), which alters glycine binding, and other amino acid residues involved in the binding of this agonist (shown in blue). 2) Hippocampal neurons infected with the human GluN1 or GluN1-S688Y subunit show that the S688Y mutation does not alter the total or surface expression of NMDA receptors. 3) Representative whole-cell patch-clamp recordings of primary hippocampal neurons show a shift in glycine binding afinity when S688Y mutation is present in the GluN1 subunit. 4) Due to long-term exposure to high concentrations of NMDA, more than 80 % of neurons expressing the GluN1 subunit die. The presence of the S688Y mutation in the GluN1 subunit has a neuroprotective effect and reduces neuronal death by 25 %.
Structural features in the glycine-binding sites of the GluN1 and GluN3A subunits regulate the surface delivery of NMDA receptors
NMDA receptors are glutamate receptors that play an essential role in mediating excitatory neurotransmission in central nervus system. We found using microscopy and electrophysiology that the structural features of the glycine-binding sites including a clinically relevant mutation in GluN subunits, are correlated with NMDA receptor trafficking to the cell surface. These findings provide important information regarding both physiological and pathophysiological processes in the mammalian brain. See more in the article Skrenkova et al. 2019.
, Barackova P, Langore E, Netolicky J, Rivas-Ramirez P, Rehakova K. The Extracellular Domains of GluN Subunits Play an Essential Role in Processing NMDA Receptors in the ER. Front. Neurosci. March 2021
, Korabecny J, Holubova K, Kleteckova L, Chvojkova M, Hakenova K, Prchal L, Novak M, Dolezal R, Hepnarova V, Svobodova B, Kucera T, Lichnerova K, Krausova B, Horak M, Vales K, Soukup O. 7-phenoxytacrine is a dually acting drug with neuroprotective efficacy in vivo. Biochem Pharmacol 2021 Feb 8;186:114460
Konecny J, Misiachna A, Hrabinova M, Pulkrabkova L, Benkova M, Prchal L, Kucera T, Kobrlova T, Finger V, Kolcheva M, Kortus S, Jun D, Valko M, Horak M, Soukup O, Korabecny J. Pursuing the Complexity of Alzheimer's Disease: Discovery of Fluoren-9-Amines as Selective Butyrylcholinesterase Inhibitors and N-Methyl-d-Aspartate Receptor Antagonists. Biomolecules 2020 Dec 22;11(1):3.
, Song J, Kortus S, Kolcheva M, Netolicky J, Hemelikova K, Kaniakova M, Hrcka Krausova B, Kucera T, Korabecny J, Suh YH, Horak M. The pathogenic S688Y mutation in the ligand-binding domain of the GluN1 subunit regulates the properties of NMDA receptors. Scientific reports 10:18576
Park DH, Park S, Song JM, Kang M, Lee S, Horak M, Suh YH. N‐linked glycosylation of the mGlu7 receptor regulates the forward trafficking and transsynaptic interaction with Elfn1. FASEB J 2020 Nov;34(11):14977-14996
, Hemelikova K, Kolcheva M, Kortus S, Kaniakova M, Krausova B, Horak M. Structural features in the glycine-binding sites of the GluN1 and GluN3A subunits regulate the surface delivery of NMDA receptors. Scientific reports 9:12303.
, Kolcheva M, Skrenkova K, Kanikova M, Horak M. Lectins modulate the functional properties of GluN1/GluN3A – containg receptors. Neuropharmacology 157, 107671.
, Nepovinova E, Kleteckova L, Skrenkova K, Chrienova Z, Hepnarova V, Vales K, Soukup O, Korabecny J*, Horak M*. Combination of Memantine and 6-Chlorotacrine as Novel Multi-Target Compound against Alzheimer’s Disease. Current Alzheimer Research 2019;16(9):821-833.
, Kleteckova L, Lichnerova K, Holubova K, Skrenkova K, Korinek M, Krusek J, Smejkalova T, Korabecny J, Vales K, Soukup O, Horak M*. 7-Methoxyderivative of tacrine is a 'foot-in-the-door' open-channel blocker of GluN1/GluN2 and GluN1/GluN3 NMDA receptors with neuroprotective activity in vivo. Neuropharmacology. 2018;140:217-232.
, Lee S, Lichnerova K, Kaniakova M, Hansikova H, Zapotocky M, Suh YH*, Horak M*. N-Glycosylation Regulates the Trafficking and Surface Mobility of GluN3A-Containing NMDA Receptors. Front Mol Neurosci. 2018 Jun 4;11:188.
Targeted and Improved Alzheimer's Disease Drug Development
Dually acting cognitive enhancers for palliative treatment of Alzheimer´s disease
AZV, NU20-08-00296, 2020-2024
Delineating the mechanisms that regulate specific NMDA receptor subtypes in mammalian neurones
GAČR, 20-12420S, 2020-2023
Novel neuroprotective compounds based on NMDA receptor antagonism and cholinergic stimulation
GAČR, 20-12047S, 2020-2023
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. and 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.