Nucleoporins (NUPs) are proteins that structure the nuclear pore complexes (NPCs). The NPC spans the nuclear envelope of a cell and provides a channel through which RNA and proteins travel between the nucleus and the cytoplasm. NUPs and NPCs disruptions have a great impact in the pathophysiology of neurodegenerative diseases1. Nup358 is a member of the large nucleoporin family that constitutes the NPCs. Nup358 interacts with specific cellular proteins and its function depends on the specific cell type expressing that nucleoporin. In developing neurons, Nup358 regulates axon specification and neuronal polarity through an interaction with other proteins2. Although at the axon initial segment (AIS) level it has been shown that when Nup358 was downregulated the signal of the AIS scaffold protein ankyrin-G was reduced3, the function of Nup358 in mature neurons remains still to be clarified. In cultures of mouse cortical neurons, we observed that the downregulation of Nup358 by shRNAs promotes an increase in neuron excitability. Using patch-clamp electrophysiological recordings, in current-clamp mode, we measured an alteration into both the active and passive properties of downregulated Nup358 neurons as compared to those exposed to a scrambled sequence. Moreover, in voltage-clamp mode, we identified clear modifications of voltage-gated sodium channel kinetics as a result of Nup358 downregulation. Taken together, our findings suggest that Nup358 exerts an active role in the regulation of neuronal excitability through a functional stabilization of the electrical properties of the neuronal membrane. 1.Coyne AN & Rothstein JD. Nat Rev Neurol. 18, 348-362 (2022). 2.Vyas P, Singh A., Murawala P & Joseph J. Biol. Open 2, 1270-1278 (2013). 3.Khalaf B, Roncador A, Pischedda F, Casini A, Thomas S, Piccoli G, Kiebler M & Macchi P. J Cell Sci. 132, jcs.222802 (2019).
PATCH-CLAMP STUDY OF NUCLEOPORIN NUP358 DOWNREGULATION IN MOUSE CORTICAL NEURONS
Carlo Musio
2022
Abstract
Nucleoporins (NUPs) are proteins that structure the nuclear pore complexes (NPCs). The NPC spans the nuclear envelope of a cell and provides a channel through which RNA and proteins travel between the nucleus and the cytoplasm. NUPs and NPCs disruptions have a great impact in the pathophysiology of neurodegenerative diseases1. Nup358 is a member of the large nucleoporin family that constitutes the NPCs. Nup358 interacts with specific cellular proteins and its function depends on the specific cell type expressing that nucleoporin. In developing neurons, Nup358 regulates axon specification and neuronal polarity through an interaction with other proteins2. Although at the axon initial segment (AIS) level it has been shown that when Nup358 was downregulated the signal of the AIS scaffold protein ankyrin-G was reduced3, the function of Nup358 in mature neurons remains still to be clarified. In cultures of mouse cortical neurons, we observed that the downregulation of Nup358 by shRNAs promotes an increase in neuron excitability. Using patch-clamp electrophysiological recordings, in current-clamp mode, we measured an alteration into both the active and passive properties of downregulated Nup358 neurons as compared to those exposed to a scrambled sequence. Moreover, in voltage-clamp mode, we identified clear modifications of voltage-gated sodium channel kinetics as a result of Nup358 downregulation. Taken together, our findings suggest that Nup358 exerts an active role in the regulation of neuronal excitability through a functional stabilization of the electrical properties of the neuronal membrane. 1.Coyne AN & Rothstein JD. Nat Rev Neurol. 18, 348-362 (2022). 2.Vyas P, Singh A., Murawala P & Joseph J. Biol. Open 2, 1270-1278 (2013). 3.Khalaf B, Roncador A, Pischedda F, Casini A, Thomas S, Piccoli G, Kiebler M & Macchi P. J Cell Sci. 132, jcs.222802 (2019).I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
