Neurosteroid modulation of NMDA receptors
N-methyl-D-aspartate (NMDA) receptors are a type of ionotropic glutamate receptors mediating fast synaptic transmission and essential for learning and memory. Abnormal function of NMDA receptors underlies many neurological and psychiatric diseases, including schizophrenia and neurodegenerative disorders. We use advanced electrophysiology, molecular biology, optogenetics, and microscopy to investigate the relationship between the structure and the function of NMDA receptor ion channels, to characterize molecular mechanisms of action of different pharmacological agents influencing them, and to study the physiological and pathological processes in which NMDA receptors participate. For details please see: http://www.fgu.cas.cz/vyklicky-vlachova-neuro/
We have identified different neurosteroid compounds with potentiating or inhibitory effects on NMDA receptors. This project will focus on a detailed study of the mechanisms of neurosteroid actions on NMDA receptors, synaptic transmission, and behavior, both under normal conditions and in selected biological models of neurological and psychiatric disorders.
Candidate Requirements: Candidates should have a Masters' degree in biological, medical or chemical subjects, or be due to complete their studies in this academic year. Experience in electrophysiology, cell biology, molecular biology, or genetics would be an advantage. Candidates will be expected to undertake independent research in their chosen project.
Petrovic MM, Viana da Silva S, Clement JP, Vyklicky L, Mulle C, Gonzalez-Gonzalez IM, Henley JM (2017) Metabotropic action of postsynaptic kainate receptors triggers hippocampal long-term potentiation. Nat Neurosci 20:529-539.
Slavikova B, Chodounska H, Nekardova M, Vyklicky V, Ladislav M, Hubalkova P, Krausova B, Vyklicky L, Kudova E (2016) Neurosteroid-like Inhibitors of N-Methyl-D-Aspartate Receptor: Substituted 2-Sulfates and 2-Hemisuccinates of Perhydrophenanthrene. Journal of medicinal chemistry 59:4724-4739.
Vyklicky V, Smejkalova T, Krausova B, Balik A, Korinek M, Borovska J, Horak M, Chvojkova M, Kleteckova L, Vales K, Cerny J, Nekardova M, Chodounska H, Kudova E, Vyklicky L. Preferential inhibition of tonically over phasically activated NMDA receptors by pregnane derivatives. J Neurosci. 36(7):2161-75 (2016).
Vyklicky V, Krausova B, Cerny J, Balik A, Zapotocky M, Novotny M, Lichnerova K, Smejkalova T, Kaniakova M, Korinek M, Petrovic M, Kacer P, Horak M, Chodounska H, Vyklicky L. Block of NMDA receptor channels by endogenous neurosteroids: implications for the agonist induced conformational states of the channel vestibule. Sci Rep. 5:10935 (2015).
Supervisors: Prof. Ladislav Vyklický, Ph.D., D.Sc.
Early processing of ionotropic glutamate receptors in rat and human neurons
N-methyl-D-aspartate receptors (NMDARs) are a subclass of glutamate receptors that play an essential role in mediating excitatory neurotransmission and synaptic plasticity in the mammalian central nervous system. Dysregulation of NMDARs plays a critical role in the aetiology of many neuropsychiatric disorders and conditions, including Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, epilepsy, anxiety, depression, bipolar disorder, schizophrenia, lupus erythematosus, and ischaemia. Thus, understanding the molecular mechanisms that regulate NMDARs is essential for designing effective therapies for these patients. In this PhD project, you will use a combination of microscopy, biochemistry, and electrophysiology to investigate the molecular mechanisms that regulate the early processing and function of NMDARs in primary rat hippocampal neurones and induced human neurons derived from fibroblasts from patients with neuropsychiatric disorders. The results obtained from these experiments will increase our knowledge regarding the mechanisms by which neurons regulate the number, type, and function of surface NMDARs.
Candidate’s profile (requirements): M.Sc. or equivalent in molecular biology, microbiology, (bio)chemistry or medicine; fascination by the complexity of neuronal functions.
● Lichnerova K, Kaniakova M, Park SP, Skrenkova K, Wang YX, Petralia RS, Suh YH, Horak M. Two N-glycosylation Sites in the GluN1 Subunit Are Essential for Releasing N-methyl-d-aspartate (NMDA) Receptors from the Endoplasmic Reticulum. Journal of Biological Chemistry 290:18379-90 (2015).
● Korinek M, Vyklicky V, Borovska J, Lichnerova K, Kaniakova M, Krausova B, Krusek J, Balik A, Smejkalova T, Horak M, Vyklicky L. Cholesterol modulates open probability and desensitization of NMDA receptors. Journal of Physiology 593:2279-93 (2015).
● Horak M, Petralia RS, Kaniakova M, Sans N. ER to synapse trafficking of NMDA receptors. Front Cell Neurosci. 8:394 (2014).
● Kaniakova M, Krausova B, Vyklicky V, Korinek M, Lichnerova K, Vyklicky L, Horak M*. Key amino acid residues within the third membrane domains of NR1 and NR2 subunits contribute to the regulation of the surface delivery of NMDA receptors. Journal of Biological Chemistry 287:26423-34 (2012).
Supervisor: Martin Horak, PhD
PhD Project 3:
The structure and dynamics of ionotropic glutamate receptor during the ion channel gating
The ionotropic glutamate receptors (AMPA and NMDA receptors) are responsible for excitatory synaptic transmission in the central nervous system and underlie the processes of learning and memory formation. Currently solved crystal and cryo-EM structures of receptor represent a significant progress in the understanding of the details of its function. However, the precise mechanism of ligand-dependent ion channel gating is still poorly understood. For a complete understanding of the process of receptor activation/deactivation is necessary to extend existing information by further studies. In this project, you will employ a combination of different methods (molecular biology, electrophysiology, protein expression, structural mass spectrometry and bioinformatics) in order to obtain data through which we will create new models of the receptor in its different functional states. Subsequently, we will determine critical structures and residues that are employed in the activation or deactivation of the receptor. The obtained results will contribute to understanding the ion channel opening and may help identify novel regulatory parts of the glutamate receptor. (This project will be co-supervised with Dr. Jiří Černý, IBT CAS)
Candidates should have a Masters' degree in biological, medical, chemical or bioinformatics subjects. Experience in molecular biology, cell biology, or structural studies would be an advantage.
Ladislav M, Cerny J, Krusek J, Skrenova K, Horak M, Balik A, Vyklicky L. Molecular insight into the N-methyl-D-aspartate receptor channel gating SfN 2017 (http://www.abstractsonline.com/pp8/#!/4376/presentation/14036)
Balik A, Penn AC, Nemoda Z, Greger IH. Activity-regulated RNA editing in select neuronal subfields in hippocampus. Nucleic Acids Res. (2013)
Penn AC, Balik A, Wozny C, Cais O, Greger IH. Activity-mediated AMPA receptor remodeling, driven by alternative splicing in the ligand-binding domain. Neuron (2012)
Supervisor: Ales Balik, PhD