IRCCS San Raffaele Scientific Institute, Via Olgettina 60, 20132, Milan, Italy.
Vita-Salute San Raffaele University, Via Olgettina 58, 20132, Milan, Italy.
Cell Death Dis. 2020 Oct 14;11(10):856. doi: 10.1038/s41419-020-03073-w.
Mutations in proline-rich transmembrane protein 2 (PRRT2) have been recently identified as the leading cause of a clinically heterogeneous group of neurological disorders sharing a paroxysmal nature, including paroxysmal kinesigenic dyskinesia and benign familial infantile seizures. To date, studies aimed at understanding its physiological functions in neurons have mainly focused on its ability to regulate neurotransmitter release and neuronal excitability. Here, we show that PRRT2 expression in non-neuronal cell lines inhibits cell motility and focal adhesion turnover, increases cell aggregation propensity, and promotes the protrusion of filopodia, all processes impinging on the actin cytoskeleton. In primary hippocampal neurons, PRRT2 silencing affects the synaptic content of filamentous actin and perturbs actin dynamics. This is accompanied by defects in the density and maturation of dendritic spines. We identified cofilin, an actin-binding protein abundantly expressed at the synaptic level, as the ultimate effector of PRRT2. Indeed, PRRT2 silencing unbalances cofilin activity leading to the formation of cofilin-actin rods along neurites. The expression of a cofilin phospho-mimetic mutant (cof-S3E) is able to rescue PRRT2-dependent defects in synapse density, spine number and morphology, but not the alterations observed in neurotransmitter release. Our data support a novel function of PRRT2 in the regulation of the synaptic actin cytoskeleton and in the formation of synaptic contacts.
富含脯氨酸的跨膜蛋白 2 (PRRT2) 的突变最近被确定为一组具有阵发性特征的神经紊乱的主要原因,包括阵发性运动诱发性运动障碍和良性家族性婴儿癫痫。迄今为止,旨在了解其在神经元中的生理功能的研究主要集中在其调节神经递质释放和神经元兴奋性的能力上。在这里,我们表明,非神经元细胞系中 PRRT2 的表达抑制细胞迁移和焦点黏附的转化,增加细胞聚集倾向,并促进丝状伪足的突出,所有这些过程都影响肌动蛋白细胞骨架。在原代海马神经元中,PRRT2 的沉默会影响丝状肌动蛋白的突触含量,并扰乱肌动蛋白动力学。这伴随着树突棘密度和成熟度的缺陷。我们确定了肌动蛋白结合蛋白 cofilin 为 PRRT2 的最终效应子,该蛋白在突触水平上大量表达。事实上,PRRT2 的沉默会破坏 cofilin 的活性,导致 cofilin-actin 杆沿着神经突形成。cofilin 磷酸模拟突变体 (cof-S3E) 的表达能够挽救 PRRT2 依赖的突触密度、棘突数量和形态的缺陷,但不能挽救神经递质释放观察到的改变。我们的数据支持 PRRT2 在调节突触肌动蛋白细胞骨架和形成突触接触中的新功能。