Funkční a farmakologické vlastnosti GluN1/Glun2 a GluN1/GluN3 podtypů NMDA receptorů

(CZ) N-methyl-D-aspartátové receptory (NMDAR) jsou ionotropní glutamátové receptory, které hrají klíčovou roli v excitačním synaptickém přenosu v centrálním nervovém systému (CNS) savců. Zvýšená ale i snížená aktivita NMDAR může způsobit široké spektrum patologických stavů a psychiatrických poruch jako například Alzheimerovu chorobu, Parkinsonovu chorobu, Huntingtonovu chorobu, epilepsii nebo schizofrenii. NMDAR tvoří heterotetramerní komplexy složené z podjednotek GluN1, GluN2(A-D) a/nebo GluN3(A, B). Různé podtypy NMDAR mohou mít rozdílný vliv na patogenezi onemocnění, a proto je zásadní zkoumat specifickou roli jednotlivých podjednotek v regulaci normálního fungování NMDAR. Regulace NMDAR probíhá na různých úrovních, od raného zpracování, včetně syntézy, sestavení, kontroly kvality v endoplazmatickém retikulu (ER) a přesunu na povrch buňky až po internalizaci, recyklaci a degradaci receptoru. V této disertační práci jsme se zaměřili především na určení role extracelulárních a transmembránových oblastí různých podtypů NMDAR na regulaci jejich funkce. Konkrétně jsme pomocí elektrofyziologických a mikroskopických metod na buňkách HEK293 a kultivovaných hipokampálních neuronech zkoumali: (i) vliv N- glykosylace a různých lektinů na regulaci funkčních vlastností receptorů GluN1/GluN3; (ii) vliv...(EN) N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors and they play a critical role in excitatory synaptic transmission in the mammalian central nervous system (CNS). Hyperactivity or hypoactivity of NMDARs can lead to a wide spectrum of pathological conditions and psychiatric disorders, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, epilepsy, schizophrenia. NMDARs form a heterotetrameric complex made up of GluN1, GluN2(A-D) and/or GluN3(A, B) subunits. Different subtypes of NMDARs could have various effects on disease pathogenesis and therefore it is crucial to investigate the specific role of each subunit in the regulation of normal NMDAR functioning. The regulation of NMDARs occurs at different levels, from early processing, including synthesis, assembly, quality control in the endoplasmic reticulum (ER), trafficking to the cell surface, to internalization, recycling, and degradation. In this dissertation, we mainly focused on determining the roles of extracellular and transmembrane regions of different subtypes of NMDARs in the regulation of their function. In particular, using electrophysiology and microscopy methods on HEK293 cells and cultured hippocampal neurons, we investigated: (i) the impact of N-glycosylation and different lectins on...Katedra fyziologieDepartment of PhysiologyPřírodovědecká fakultaFaculty of Scienc

Functional and pharmacological properties of GluN1/GluN2 and GluN1/GluN3 subtypes of NMDA receptors

(EN) N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors and they play a critical role in excitatory synaptic transmission in the mammalian central nervous system (CNS). Hyperactivity or hypoactivity of NMDARs can lead to a wide spectrum of pathological conditions and psychiatric disorders, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, epilepsy, schizophrenia. NMDARs form a heterotetrameric complex made up of GluN1, GluN2(A-D) and/or GluN3(A, B) subunits. Different subtypes of NMDARs could have various effects on disease pathogenesis and therefore it is crucial to investigate the specific role of each subunit in the regulation of normal NMDAR functioning. The regulation of NMDARs occurs at different levels, from early processing, including synthesis, assembly, quality control in the endoplasmic reticulum (ER), trafficking to the cell surface, to internalization, recycling, and degradation. In this dissertation, we mainly focused on determining the roles of extracellular and transmembrane regions of different subtypes of NMDARs in the regulation of their function. In particular, using electrophysiology and microscopy methods on HEK293 cells and cultured hippocampal neurons, we investigated: (i) the impact of N-glycosylation and different lectins on..