11.1. 2016, Buněčná neurofyziologie

PhD Project1: 

Early processing of ionotropic glutamate receptors in rat and human neurons

 

Research Summary:

 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 Requirements:

M.Sc. or equivalent in molecular biology, microbiology, (bio)chemistry or medicine; fascination by the complexity of neuronal functions.

Relevant publications:

● 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).

 Supervisors: Martin Horak, PhD  

 

 

PhD Project2: 

Neurosteroid modulation of NMDA receptors

Research Summary:

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.biomed.cas.cz/odd031/researchlv.html

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 cell biology, molecular biology, or genetics would be an advantage.  Candidates will be expected to undertake independent research in their chosen project.

Relevant Publications:

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).

Kudova E, Chodounska H, Slavikova B, Budesinsky M, Nekardova M, Vyklicky V, Krausova B, Svehla P, Vyklicky L. A New Class of Potent N-Methyl-D-Aspartate Receptor Inhibitors: Sulfated Neuroactive Steroids with Lipophilic D-Ring Modifications. J Med Chem. 58(15):5950-66 (2015).

Supervisors:  Prof. Ladislav Vyklický, Ph.D., D.Sc.

 

PhD Project3: 

The role of  iGluR complex in the emergence and development of  schizophrenia.  

 

Research Summary:

Ionotropic glutamate receptors (iGluRs) mediate excitatory synaptic transmission and are essential for processes of learning and memory formation. Dysfunction of AMPA and NMDA receptors underlies many neurological disorders including schizophrenia. Schizophrenia is a psychiatric disorder with a serious impact on a patient's life. We aim to determine how gene variants of the glutamate receptor complex (excitatory synapse) contribute to the emergence and development of schizophrenia. We employ NGS, bioinformatics, structural and molecular biology and electrophysiology techniques to characterize the functional consequences of altered glutamate receptor complex gene variants. This project will contribute to the identification of potential cellular markers of schizophrenia.

Candidate Requirements:

Candidates should have a Masters' degree in biological, medical, chemical or bioinformatics subjects. Experience in molecular biology, cell biology, or human genetics would be an advantage. 

Publications:

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

 

PhD Project4:

The glutamate NMDA receptor as a target of novel drugs for the treatment of Alzheimer disease

 

Research Summary:

1,2,3,4-tetrahydro-9-aminoacridine (THA) was the first approved (by FDA in 1993) multitargeted drug for the Alzheimer disease (AD) treatment. In the collaborative project with the Biomedical Research Center in Hradec Kralove and the National Institute of Mental Health in Klecany, you will help to design and characterize novel THA-based compounds acting at multiple targets, including the glutamate type of the N-methyl-D-aspartate receptors (NMDARs). Specifically, you will compare the effectivenessness of the FDA-approved and novel drugs on the NMDARs and the AD-related alterations, by performing:

i.) whole-cell and single-channel patch-clamp recordings from the mammalian cell lines expressing the recombinant NMDARs,

ii.) synaptic recordings ("NMDAR-EPSC") from the cultured hippocampal neurons grown on the microislands,

iii.) biochemical and microscopical analysis of the hippocampal neurons infected with the shRNA-expressing lentiviruses and the human neurons derived from the fibroblasts of the AD patients,

iv.) behavioral studies on the AD animal models.

Candidate’s profile (requirements):

M.Sc. or equivalent in molecular biology, microbiology, (bio)chemistry or medicine; motivation to find novel drugs with unique mechanism of action.

Publications:

• Horak M, Benkovicova K, Krusek J, Kaniakova M, Korabecny J, Nepovimova E, Vyklicky L, Kuca K, Stuchlik A, Ricny J, Vales K, Soukup O. The pharmacology of tacrine at N-methyl-D-aspartate receptors. Progress in Neuropsychopharmacology & Biological Psychiatry 75:54-62 (2017).

• Gazova Z, Soukup O, Sepsova V, Siposova K, Drtinova L, Jost P, Spilovska K, Korabecny J, Nepovimova E, Fedunova D, Horak M, Kaniakova M, Wang ZJ, Hamouda AK, Kuca K. Multi-target-directed therapeutic potential of 7-methoxytacrine-adamantylamine heterodimers in the Alzheimer's disease treatment. Biochim Biophys Acta. 1863(2):607-619 (2017).

• 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. J Biol Chem. 290:18379-90 (2015).

Supervisor: Martin Horak, PhD