| Popis: |
The basal ganglia play an essential role in movement selection and control. It is therefore not surprising that they are central to neurodegenerative diseases like Parkinson’s disease and Huntington’s disease. As such, greater knowledge of how the different parts of the basal ganglia behave, both in their normal and in disease states, will increase our understanding of the system and help in finding new and improved ways to treat the possible disorders.In this thesis we were interested in the starting point of the basal ganglia, the medium spiny neurons of the striatum. Generally categorized into two groups, the direct pathway medium spiny neurons, projecting to the substantia nigra and responsible for activating action, and the indirect pathway medium spiny neurons, projecting to the globus pallidus and responsible for inhibiting action. We were interested in exploring their respective miRNA profiles to find if there were large differences to be found there. MiRNA are short RNA, around 22 nt in length that function as guides for the RNA-induced silencing complex to downregulate protein expression. Through the expression of a fusion-protein composed of one of the components of the RNA-induced silencing complex responsible for miRNA binding, and GFP, it is possible to isolate the miRNA populations of cells expressing this fusion-protein. We injected virus, expressing the fusion-protein and capable of retrograde transport, into these areas. We were able to show that both a lentivirus containing a rabies virus fusion envelope, and an adeno-associated virus with a modified AAV2 capsid could be used for this purpose. Following successful transduction of cells in the striatum by retrograde transport, miRNA was sequenced and compared between the two populations. In total eight different miRNA were found to be differentially expressed between them. One of which had simultaneously also been identified by another group using a different set of methods in mice. Finally, we also explore the expression profile of a dopamine receptor D2 specific promoter element within the dorsal striatum. |
