Zebra balığında glutamat sinyalizasyon manipülasyonunun oligodendrosit kökenli hücrelerdeki etkisinin incelenmesi
Özet
Myelination is crucial for development and maintaining axonal integrity. There is increasing evidence that glutamate-dependent neuronal activity may modulate the myelination process. The aim of this project is to investigate the effect of manipulating glutamate signaling on zebrafish oligodendrocyte lineage cells. A newly designed "intensity-based glutamate-sensing fluorescent reporter" (iGluSnFr) zebra fish model is used to investigate the effect of manipulating glutamate signaling on myelinating oligodendrocytes. According to our hypothesis iGluSnFR would bind and thereby buffers away extracellular Glu. By reducing extracellular Glu levels, OL progenitors lack some cues for proliferation and differentiation. This reduces in the end OL lineage cell numbers. For this purpose the expression of myelin markers were evaluated in zebra-fish expressing iGluSnFR compared to controls. To verify that iGluSnFR is responsible for the reduction of oligodendrocyte lineage cells considering that the glutamate related pharmacological tools may not cross the blood-brain barrier and reach their goals micro-injections were applied into the brain ventricles of the larvae zebra fish. Finally myelination was evaluated by in vivo imaging, qRT-PCR and WB techniques.