A5009975726- Neuroscience and Neuropharmacology Research
- Receptor Mechanisms and Signaling
- Ion channel regulation and function
- X-ray Diffraction in Crystallography
- Crystallization and Solubility Studies
- Nanoplatforms for cancer theranostics
- Photodynamic Therapy Research Studies
- Neuroendocrine regulation and behavior
- Estrogen and related hormone effects
- Photoreceptor and optogenetics research
- Molecular Sensors and Ion Detection
- Lipid Membrane Structure and Behavior
- Genetics and Neurodevelopmental Disorders
- Porphyrin and Phthalocyanine Chemistry
- Stress Responses and Cortisol
- Neuroinflammation and Neurodegeneration Mechanisms
- Photochromic and Fluorescence Chemistry
- Bioactive Compounds and Antitumor Agents
- Luminescence Properties of Advanced Materials
- Medicinal Plant Pharmacodynamics Research
- Education, Psychology, and Social Research
- Luminescence and Fluorescent Materials
- Biochemical effects in animals
- Synthesis and Reactivity of Sulfur-Containing Compounds
- Memory and Neural Mechanisms
University of California, Berkeley
2019-2024
Czech Academy of Sciences, Institute of Physiology
2012-2023
Czech Academy of Sciences
2011-2018
Charles University
2008-2015
Key points NMDA receptors (NMDARs) are tetrameric cation channels permeable to calcium; they mediate excitatory synaptic transmission in the CNS and their excessive activation can lead neurodegeneration. Although these direct contact with plasma membrane, lipid–NMDAR interactions little understood. Using cultured rat cerebellar granule cells, we show that acute chronic pretreatments resulting cell cholesterol depletion profoundly diminish NMDAR responses increase desensitization, also...
G protein-coupled receptors (GPCRs) are membrane proteins that play important roles in biology. However, our understanding of their function complex living systems is limited because we lack tools can target individual with sufficient precision. State-of-the-art approaches, including DREADDs, optoXRs, and PORTL gated-receptors, control GPCR signaling molecular, cell type, temporal specificity. Nonetheless, these based on engineered non-native may (i) express at nonphysiological levels, (ii)...
Metabotropic glutamate receptors (mGluRs) are dimeric G-protein-coupled that operate at synapses. Macroscopic and single molecule FRET to monitor structural rearrangements in the ligand binding domain (LBD) of mGluR7/7 homodimer revealed it have an apparent affinity ~4000-fold lower than other mGluRs a maximal activation only ~10%, seemingly too low for However, mGluR7 heterodimerizes, we find associate with mGluR2 hippocampus. Strikingly, mGluR2/7 heterodimer has high efficacy. shows...
N-methyl-D-aspartate receptors (NMDARs), glutamate-gated ion channels, mediate signaling at the majority of excitatory synapses in nervous system. Recent sequencing data for neurological and psychiatric patients have indicated numerous mutations genes encoding NMDAR subunits. Here, we present surface expression, functional, pharmacological analysis 11 de novo missense human hGluN2B subunit (P553L; V558I; W607C; N615I; V618G; S628F; E657G; G820E; G820A; M824R; L825V) located preM1, M1, M2,...
Abstract N-methyl-D-aspartate receptors (NMDARs) mediate synaptic plasticity and their dysfunction is implicated in multiple brain disorders. NMDARs can be allosterically modulated by numerous compounds, including endogenous neurosteroid pregnanolone sulfate. Here, we identify the molecular basis of use-dependent voltage-independent inhibitory effect neurosteroids on NMDAR responses. The site action located at extracellular vestibule receptor’s ion channel pore accessible after receptor...
Postsynaptic N -methyl- d -aspartate receptors (NMDARs) phasically activated by presynaptically released glutamate are critical for synaptic transmission and plasticity. However, under pathological conditions, excessive activation of NMDARs tonically increased ambient contributes to excitotoxicity associated with various acute chronic neurological disorders. Here, using heterologously expressed GluN1/GluN2A GluN1/GluN2B rat autaptic hippocampal microisland cultures, we show that pregnanolone...
BACKGROUND AND PURPOSE NMDA receptors are glutamatergic ionotropic involved in excitatory neurotransmission, synaptic plasticity and excitotoxic cell death. Many allosteric modulators can influence the activity of these positively or negatively, with behavioural consequences. 20‐Oxo‐5β‐pregnan‐3α‐yl sulphate (pregnanolone sulphate; PA‐6) is an endogenous neurosteroid that inhibits neuroprotective. We tested hypothesis interaction PA‐6 plasma membrane critical for its inhibitory effect at...
Highlights•smFRET reveals conformational differences between NMDA receptors with different GluN2s•Low-Po are more splayed at rest and compact when fully agonized•Interaction the GluN1 LBD GluN2 NTD contributes to these differencesSummaryN-methyl-D-aspartate ionotropic glutamate that mediate synaptic transmission plasticity. Variable subunits in diheterotetrameric identical set very functional properties. To understand this diversity, we use single-molecule fluorescence resonance energy...
N-Methyl-D-aspartate receptors (NMDARs) are glutamate-gated ion channels that play a crucial role in excitatory synaptic transmission. However, the overactivation of NMDARs can lead to excitotoxic cell damage/death, and as such, they numerous neuropathological conditions. The activity is known be influenced by wide variety allosteric modulators, including neurosteroids, which turn makes them promising therapeutic targets. In this study, we describe new class neurosteroid analogues possess...
<i>N</i>-methyl-D-aspartate receptor (NMDAR) hypofunction has been implicated in several neurodevelopmental disorders. NMDAR function can be augmented by positive allosteric modulators, including endogenous compounds, such as cholesterol and neurosteroid pregnenolone sulfate (PES). Here we report that PES accesses the via membrane, its binding site is different from of cholesterol. Alanine mutagenesis identified residues disrupt steroid potentiating effect at rat GluN1 (G638; I642) GluN2B...
Abstract N-Methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate essential for synaptic plasticity and memory. Receptor activation involves glycine- glutamate-stabilized closure of the GluN1 GluN2 subunit ligand binding domains that is allosterically regulated by amino-terminal domain (ATD). Using single molecule fluorescence resonance energy transfer (smFRET) to monitor rearrangements in real-time, we observe a stable ATD inter-dimer distance Apo state test effects agonists...
N-methyl-D-aspartate receptors are ionotropic glutamate that integral to synaptic transmission and plasticity. Variable GluN2 subunits in diheterotetrameric with identical GluN1 set very different functional properties, which support their individual physiological roles the nervous system. To understand conformational basis of this diversity, we assessed conformation common subunit using single-molecule fluorescence resonance energy transfer (smFRET). We established smFRET sensors ligand...
A unique asymmetric total synthesis of the unnatural enantiomer pregnanolone, as well a study its biological activity at NMDA receptor, is reported. The asymmetry introduced by highly atom-economic organocatalytic Robinson annulation. new method for construction cyclopentane D-ring consisting CuI-catalyzed conjugate addition and oxygenation followed thermal cyclization employing persistent radical effect was developed. ent-Pregnanolone sulfate surprisingly only 2.6-fold less active than...
NMDARs are ligand-gated ion channels that cause an influx of Na + and Ca 2+ into postsynaptic neurons. The resulting intracellular transient triggers synaptic plasticity. When prolonged, it may induce excitotoxicity, but also activate negative feedback to control the activity NMDARs. Here, we report a rise in (Ca challenge) increases sensitivity not AMPARs/kainate receptors endogenous inhibitory neurosteroid 20-oxo-5β-pregnan-3α-yl 3-sulfate its synthetic analogs, such as 3-hemipimelate...
Pregnenolone sulfate (PS), an endogenously occurring neurosteroid, has been shown to modulate the activity of several neurotransmitter-gated channels, including NMDA receptor (NMDAR). NMDARs are glutamate-gated ion channels involved in excitatory synaptic transmission, plasticity, and excitotoxicity. In this study, we analyzed effects PS on calcium signaling cultured hippocampal neurons HEK293 cells expressing NMDAR. The were loaded with Ca2+ sensor Fura-2. agreement previous...
Here, we report the synthesis of pregn-5-ene and androst-5-ene dicarboxylic acid esters explore structure–activity relationship (SAR) for their modulation N-methyl-d-aspartate receptors (NMDARs). All compounds were positive modulators recombinant GluN1/GluN2B (EC50 varying from 1.8 to 151.4 μM Emax 48% 452%). Moreover, 10 found be more potent receptor than endogenous pregnenolone sulfate = 21.7 μM). The SAR study revealed a between length residues at carbon C-3 steroid molecule modulatory...
Background and Purpose N‐methyl‐D‐aspartate receptors (NMDARs) play a critical role in synaptic plasticity, mutations human genes encoding NMDAR subunits have been described individuals with various neuropsychiatric disorders. Compounds positive allosteric effect are thought to compensate for reduced receptor function. Experimental Approach We used whole‐cell patch‐clamp electrophysiology on recombinant rat NMDARs variants found disorders, combination silico modelling, explore the site of...
Multiple molecular targets have been identified to mediate membrane-delimited and nongenomic effects of natural synthetic steroids, but the influence steroid metabolism on neuroactive signaling is not well understood. To begin address this question, we set out identify major metabolites a neuroprotective 20-oxo-5β-pregnan-3α-yl l-glutamyl 1-ester (pregnanolone glutamate, PAG) characterize their GABAA NMDA receptors (GABARs, NMDARs) zebrafish behavior. Gas chromatography–mass spectrometry was...
N-Methyl-d-aspartate receptors (NMDARs) display a critical role in various diseases of the central nervous system. The activity NMDARs can be modulated by neurosteroids. Herein, we report structure-activity relationship study for perhydrophenanthrene analogues possessing framework that mimics steroidal ring This comprises design, synthesis, and assessment biological library 2-sulfates 2-hemisuccinates (1-10). Their ability to modulate NMDAR-induced currents was tested on recombinant...