Rekombinantni GABA-A receptori - model in vitro za istraživanje neurosteroida dehidroepiandrosteron sulfata

master's thesis

• Title (english): Recombinant GABA-A receptors - in vitro model to study the effects of neurosteroid dehydroepiandrosterone sulphate
• Author: Lucija Tudor
• Mentor: Dubravka Švob Štrac (mentor)
• Mentor: Vesna Benković (mentor)
• Committee member: Vesna Benković (predsjednik povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Inga Marijanović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2016-03-15, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Dehidroepiandrosteron-sulfat (DHEAS) je neurosteroid uključen u važne funkcije mozga poput neuralne plastičnosti, učenja, pamćenja i ponašanja, te pokazuje potencijal u liječenju različitih neuropsihijatrijskih poremećaja, uključujući i ishemijski moždani udar. Učinci DHEAS-a u središnjem živčanom sustavu posredovani su putem GABA i glutamatnog neurotransmitorskog sustava, ali i drugih. Cilj rada bio je ispitati učinke primjene DHEAS-a na rekombinantne GABAA receptore, usporedbom djelovanja ovog neurosteorida na embrionalne stanice bubrega čovjeka (HEK 293), netransficirane i stabilno transficirane GABAA receptorima 122S, te istražiti njegovo potencijalno neuroprotektivno djelovanje u modelu deprivacije kisika i glukoze, te reperfuzije in vitro (OGDR). Istraživanje je pokazalo da se DHEAS ponaša kao alosterički antagonist rekombinantnih GABAA receptora, ali da njegova produljena primjena ne utječe na broj, afinitet, kao ni na funkcionalnu povezanost receptorskih veznih mjesta. Rezultati su također pokazali da DHEAS, primijenjen i prije i nakon OGDR djeluje protektivno na stanice HEK 293, te da se citoprotektivan učinak ovog neurosteroida barem dijelom ostvaruje putem GABAA receptora.

Abstract (english)

Dehydroepiandrosterone sulfate (DHEAS) is a neurosteroid involved in various important brain functions such as neural plasticity, learning, memory and behavior, demonstrating the potential to treat a variety of neuropsychiatric disorders, including ischemic stroke. Effects of DHEAS in the central nervous system are mediated through GABAergic, glutamatergic and other neurotransmitter systems. The aim of this study was to investigate the effects of DHEAS on recombinant GABAA receptors by comparing the effects of this neurosteorid on human embryonic kidney (HEK) 293 cells, stably transfected and untransfected with 122S GABAA receptors, and to explore the potential neuroprotective effect of this neurosteroid using in vitro model of oxygen and glucose deprivation and reperfusion (OGDR). The study has shown that DHEAS acts as an allosteric antagonist of recombinant GABAA receptors, but that its prolonged administration does not affect the number, affinity, or the functional coupling of receptor binding sites. The results also suggested that DHEAS, applied both before and after OGDR has protective effect on HEK 293 cells, and that cytoprotective effect of this neurosteroid is at least partly achieved via GABAA receptors.

• Keywords: Dehidroepiandrosteron-sulfat (DHEAS); embrionalne stanice bubrega čovjeka (HEK 293); rekombinananti GABAA receptori; model deprivacije kisika i glukoze; te reperfuzije (OGDR); citoptotektivan učinak; ishemijska ozljeda mozga
• Keywords (english): Dehydroepiandrosterone sulfate (DHEAS); human embryonic kidney (HEK) 293 cells; recombinant GABAA receptors; model of oxygen and glucose deprivation and reperfusion (OGDR); cytoprotective effect; ishemic brain injury
• Language: croatian
• URN:NBN: urn:nbn:hr:217:197733
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-05-12 11:46:15