跳转至内容
Merck
  • The membrane periodic skeleton is an actomyosin network that regulates axonal diameter and conduction.

The membrane periodic skeleton is an actomyosin network that regulates axonal diameter and conduction.

eLife (2020-03-21)
Ana Rita Costa, Sara C Sousa, Rita Pinto-Costa, José C Mateus, Cátia Df Lopes, Ana Catarina Costa, David Rosa, Diana Machado, Luis Pajuelo, Xuewei Wang, Feng-Quan Zhou, António J Pereira, Paula Sampaio, Boris Y Rubinstein, Inês Mendes Pinto, Marko Lampe, Paulo Aguiar, Monica M Sousa
摘要

Neurons have a membrane periodic skeleton (MPS) composed of actin rings interconnected by spectrin. Here, combining chemical and genetic gain- and loss-of-function assays, we show that in rat hippocampal neurons the MPS is an actomyosin network that controls axonal expansion and contraction. Using super-resolution microscopy, we analyzed the localization of axonal non-muscle myosin II (NMII). We show that active NMII light chains are colocalized with actin rings and organized in a circular periodic manner throughout the axon shaft. In contrast, NMII heavy chains are mostly positioned along the longitudinal axonal axis, being able to crosslink adjacent rings. NMII filaments can play contractile or scaffolding roles determined by their position relative to actin rings and activation state. We also show that MPS destabilization through NMII inactivation affects axonal electrophysiology, increasing action potential conduction velocity. In summary, our findings open new perspectives on axon diameter regulation, with important implications in neuronal biology.

材料
货号
品牌
产品描述

Sigma-Aldrich
层粘连蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
杜氏改良 Eagle 培养基 - 高葡萄糖, With 4500 mg/L glucose, L-glutamine, sodium pyruvate, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
视黄酸, ≥98% (HPLC), powder
Sigma-Aldrich
杜氏改良Eagle 培养基/营养混合物 F-12 Ham, With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
胰蛋白酶 来源于猪胰腺, lyophilized powder, BioReagent, suitable for cell culture, 1,000-2,000 BAEE units/mg solid
Sigma-Aldrich
葡萄糖氧化酶 来源于黑曲霉, Type X-S, lyophilized powder, 100,000-250,000 units/g solid (without added oxygen)
Sigma-Aldrich
单克隆抗 β-肌动蛋白抗体 小鼠抗, clone AC-15, ascites fluid
Sigma-Aldrich
聚-L-赖氨酸 氢溴酸盐, mol wt 30,000-70,000
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
聚-D-赖氨酸 氢溴酸盐, mol wt 70,000-150,000
Sigma-Aldrich
正钒酸钠, ≥90% (titration)
Sigma-Aldrich
抗-α-微管蛋白抗体,小鼠单克隆, clone DM1A, purified from hybridoma cell culture
Sigma-Aldrich
脑源性神经营养因子 人, BDNF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
半胱胺, ~95%
Sigma-Aldrich
(−)-II型肌球蛋白, solid, synthetic
Sigma-Aldrich
Triton X-100, BioXtra
Sigma-Aldrich
葡萄糖 溶液, BioUltra, for molecular biology, ~20% in H2O
Sigma-Aldrich
辛酸甲酯, 99%
Sigma-Aldrich
花萼海绵诱癌素A 来源于花萼国皮海绵, ≥90% (HPLC), solid
Sigma-Aldrich
过氧化氢酶 来源于谷氨酸棒状杆菌, solution, deep brown, ≥500000 U/mL
Sigma-Aldrich
抗-肌球蛋白IIA,非肌肉 兔抗, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
ML-7, powder
Sigma-Aldrich
Anti-Myosin IIB, non-muscle antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution