Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2019
Erindi/veggspjald / Talk/poster V7
Höfundar / Authors: Fatich Mechmet (1), Eiríkur Steingrímsson (2), Pétur Henry Petersen (1)
Starfsvettvangur / Affiliations: 1. Department of Anatomy, Biomedical Center, Faculty of Medicine, School of Health Sciences, University of Iceland, 2. Department of Biochemistry and Molecular Biology, Biomedical Center, Faculty of Medicine, School of Health Sciences, University of Iceland
Kynnir / Presenter: Fatich Mechmet
Neuroplasticity, the capacity of neurons to reorganize and remodel themselves in response to intrinsic and extrinsic factors, can be divided into intrinsic or synaptic plasticity. The Microphthalmia-associated transcription factor (Mitf), a key regulator in melanocytes and melanoma development, is expressed in the projection neurons of the olfactory bulb (OB), the tufted and mitral cells. Recently, we have shown that Mitf links neuronal activity with intrinsic homeostatic neuroplasticity in the OB. The focus of this study is understanding how neuronal activity affects the MITF protein. For this purpose, mouse neuroblastoma cell line (N2A) expressing an inducible mMITF-M-FLAG-HA construct, was differentiated towards a post-mitotic neuronal fate and, treated with forskolin in a time-course experiment. Forskolin is known to induce high levels of cyclic AMP (cAMP). Subsequently, cycloheximide (CHX) - chase assay was performed in cells treated with forskolin at different time points to measure changes in the half-life of the MITF protein. Proteins were measured using western blotting. The results show that forskolin increased MITF-M lower band amount gradually and the CHX-chase experiment showed a trend of increased MITF stability in N2A cells after forskolin treatment. These results show that proxies of neuronal activity can lead to changes in the levels of the MITF protein, likely at the level of protein stability.