Title Utjecaj klorpromazina na izražaj CD81 proteina u staničnim linijama središnjeg živčanog sustava Title (english) The Effects of Chlorpromazine on the Expression of the Tetraspanin Protein CD81 in Neural Cell Lines Author Vedrana Kauzlarić Mentor Kristina Grabušić (mentor)Mentor Janja Kuharić (mentor)Committee member Mladenka Malenica Staver (predsjednik povjerenstva)Committee member Natalia Kučić (član povjerenstva)Committee member Kristina Grabušić (član povjerenstva)Committee member Janja Kuharić (član povjerenstva)Granter University of Rijeka Defense date and country 2019-09-23, Croatia Scientific / art field, BIOTECHNICAL SCIENCES Abstract U ovom radu ispitan je utjecaj antipsihotika klorpromazina na izražaj proteina CD81 u humanim stanicama porijekla iz središnjeg živčanog sustava. Klorpromazin je neuroleptik koji djeluje kao snažan antagonist dopaminskih receptora, koji sudjeluju u procesima poput učenja, pamćenja, motivacije, užitka i dr. Povrh toga, klorpromazin utječe na mnoge druge membranske i citosolne proteine što rezultira smanjenom komunikacijom između neurona i glija stanica, prvenstveno astrocita. Međutim, dosad nije opisano utječe li klorpromazin i na specifičan oblik međustanične komunikacije koji uključuje izvanstanične vezikule (IV-e). IV-e su nanostrukture omeđene membranom koje sadrže proteine, nukleinske kiseline, lipide i druge molekule. Izlučuju ih sve vrste stanica i njima sekretorne stanice mogu utjecati na fenotip primateljskih stanica. CD81 je član obitelji tetraspanina, transmembranskih proteina izraženih u svim tkivima, pa tako i u stanicama središnjeg živčanog sustava. Osim na plazmatskim membranama, CD81 prisutan je i na citosolnim membranskim odjeljcima te je prepoznat kao marker izvanstaničnih vezikula. U živčanom je sustavu CD81 uključen u interakcije neurona i astrocita te u odgovor astrocita i mikroglija-stanica na ozljedu i bolest.
U ovom radu stanične linije humanog glioblastoma (U-87 MG) i neuroblastoma (SH-SY5Y) tretirane su rastućim koncentracijama klorpromazina (4, 8, 16 i 32 µg/ml) tijekom 24 sata. Analiza protočnom citometrijom pokazala je da klorpromazin ima oprečno djelovanje na razine CD81 u ispitanim staničnim linijama. U stanicama U-87 MG dolazi do dozno-ovisnog povećanja CD81, dok kod SH-SY5Y stanica razina ovog proteina opada s porastom koncentracije klorpromazina. Ova saznanja prvi put povezuju klorpromazin i promjene izražaja CD81. Imajući na umu brojne uloge CD81 u interakcijama neurona i astrocita te prisutnost tog proteina na izvanstaničnim vezikulama, buduća istraživanja trebala bi podrobnije ispitati posljedice ovih promjena na procese međustanične komunikacije u živčanom sustavu.
Abstract (english) This study examined the effects of chlorpromazine on the expression of CD81 in representative human cell lines of the central nervous system origin. Chlorpromazine is a neuroleptic that acts as a strong antagonist of dopamine receptors, which participate in processes like learning, memory, motivation, pleasure etc. Also, chlorpromazine affects many other membrane and cytosolic proteins, resulting in decreased communication between neurons and glial cells, especially astrocytes. Nevertheless, it has not yet been determined if chlorpromazine affects a specific form of intercellular communication that involves extracellular vesicles (EVs). EVs are membranous nanostructures that carry proteins, nucleic acids, lipids and other molecules. All types of cells release EVs and with them secretory cells can change the phenotype of recipient cells. The CD81 molecule is a member of the tetraspanin protein family ubiquitously expressed in all tissues, including the central nervous system. Next to the expression in plasma membrane, CD81 is present in intracellular membrane compartments and it is a recognized marker of EVs. In the nervous system, this protein is important for neuron-astrocyte interactions and is implicated in the response of astrocytes and microglia to trauma and disease.
In this study, the human glioblastoma cell line U-87 MG and neuroblastoma SH-SY5Y cells were treated with increasing concentrations of chlorpromazine (4, 8, 16 and 32 µg/ml) for 24 hours. Flow cytometry analysis of the CD81 expression has shown that chlorpromazine affects the examined cell lines in different ways. U-87 MG cells responded to chlorpromazine with a dose-dependent increase of CD81 protein level, while SH-SY5Y cells showed decreased CD81 levels. These findings are the first to date that associate chlorpromazine with changes in CD81 expression. Considering the role of this protein in neuron-astrocyte interactions and its presence in EVs, future studies are necessary to examine the effects of chlorpromazine treatment on processes important for intercellular communication in the nervous system.
Keywords
klorpromazin
CD81
međustanična komunikacija
protočna citometrija
Keywords (english)
chlorpromazine
CD81
intercellular communication
flow cytometry
Language english URN:NBN urn:nbn:hr:193:312382 Study programme Title: Biotechnology in medicine Type of resource Text File origin Born digital Access conditions Open access Terms of use Created on 2019-09-22 20:44:22
