Izražaj gena Stam2 u živčanom sustavu miša određen na razini mRNA, bjelančevine, te pomoću lacZ reporterskog sustava [Stam2 expression in the mouse nervous system at the mRNA and protein level and using lacZ reporter system]

Kapuralin, Katarina (2012) Izražaj gena Stam2 u živčanom sustavu miša određen na razini mRNA, bjelančevine, te pomoću lacZ reporterskog sustava [Stam2 expression in the mouse nervous system at the mRNA and protein level and using lacZ reporter system]. PhD thesis, Sveučilište u Zagrebu.

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Abstract

STAM2 is a member of the endosome-associated complex ESCRT-0, and is involved in the sorting process of mono-ubiquitinated endosomal cargo for degradation in the lysosome. As STAM2 is phosphorylated upon binding diverse growth factors and cytokines to the cell membrane, it might also play a regulatory role in cell signalling, being at the intercross of signalling pathways and membrane transport in the cell. The aim of this research was to determine Stam2 expression in mouse nervous system in order to get insight into its possible role. In order to investigate Stam2 molecule, in situ RNA hybridization and immunostaining were performed. Stam2 is expressed in the central and peripheral nervous system. It was confirmed that expression pattern of Stam2 can be assessed by histochemical detection of beta-galactosidase activity in a mouse carrying a gene trap modification of Stam2 gene as well. The majority of Stam2-positive cells co-stained with the neuronal markers, indicating that Stam2-expressing cells are largely neurons. Apart in the cytosolic fraction, significant amount of STAM2 is present in the nuclear fraction as well, indicating possible dual role of STAM2 in the different subcellular compartments. This work indicates that localization of STAM2 enables endosome mediated intracellular signaling in neurons and hence involvement in the crucial events in the nervous system.

Abstract in Croatian

STAM2 dio je endosomskog kompleksa ESCRT-0, u kojem je uključen u razvrstavanje monoubikvitiniranih bjelančevina namijenjenih razgradnji u lizosomima. Budući da je STAM2 fosforiliran nakon vezanja čimbenika rasta i citokina za stanicu, sudjeluje u staničnoj signalizaciji, nalazeći se tako na križanju signalnih putova i membranskog transporta stanice. Cilj ovog rada bio je određivanje izražaja Stam2 u živčanom sustavu miša kako bi se dobio uvid u moguću ulogu toga gena u živčanom sustavu. Kako bi se istražila molekula STAM2 učinjene su tehnike hibridizacije RNA in situ i imunobojanja. Stam2 je izražen u centralnom i u perifernom živčanom sustavu. Potvrđeno je kako uzorak izražaja Stam2 također može biti praćen histokemijskim određivanjem aktivnosti β-galaktozidaze u miševa koji nose preinaku u genu Stam2. Većina Stam2-pozitivnih stanica kolokalizira s biljezima živčanih stanica, što upućuje na činjenicu da su stanice koje izražavaju Stam2 uglavnom živčane stanice. Osim u citosolnoj frakciji, značajna količina STAM2 prisutna je također i u frakciji jezgara, što upućuje na moguću dvojaku ulogu STAM2 u različitim staničnim odjeljcima. Ovaj rad ukazuje kako smještaj STAM2 u živčanim stanicama omugućuje unutarstaničnu signalizaciju posredovanu endosomima, a time prisutnost u ključnim zbivanjima u djelovanju živčanog sustava.

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