Evans L L, Hammer J, Bridgman P C
Department of Anatomy and Neurobiology, Washington University, St Louis, Missouri 63110, USA.
J Cell Sci. 1997 Feb;110 ( Pt 4):439-49. doi: 10.1242/jcs.110.4.439.
Myosin V-null mice (dilute-lethal mutants) exhibit apparent neurological defects that worsen from birth until death in the third postnatal week. Although myosin V is enriched in brain, the neuronal function of myosin V is unclear and the underlying cause of the neurological defects in these mice is unknown. To aide in understanding myosin V function, we examined the distribution of myosin V in the rodent superior cervical ganglion (SCG) growth cone, a well characterized neuronal structure in which myosin V is concentrated. Using affinity purified, myosin V-specific antibodies in immunofluorescence and immunoelectron microscopy, we observed that myosin V is concentrated in organelle-rich regions of the growth cone. Myosin V is present on a distinct population of small (50-100 nm) organelles, and on actin filaments and the plasma membrane. Myosin V-associated organelles are present on both microtubules and actin filaments. These results indicate that myosin V may be carried as a passenger on organelles that are transported along microtubules, and that these organelles may also be capable of movement along actin filaments. In addition, we found no abnormalities in outgrowth, morphology, or cytoskeletal organization of SCG growth cones from dilute-lethal mice. These results indicate that myosin V is not necessary for the traction force needed for growth cone locomotion, for organization of the actin cytoskeleton, or for filopodial dynamics.
肌球蛋白V基因敲除小鼠(稀释致死突变体)表现出明显的神经缺陷,从出生到出生后第三周死亡,这些缺陷会逐渐恶化。尽管肌球蛋白V在脑中含量丰富,但其神经元功能尚不清楚,这些小鼠神经缺陷的潜在原因也未知。为了帮助理解肌球蛋白V的功能,我们研究了肌球蛋白V在啮齿动物颈上神经节(SCG)生长锥中的分布,生长锥是一种特征明确的神经元结构,其中肌球蛋白V高度集中。通过在免疫荧光和免疫电子显微镜中使用亲和纯化的、肌球蛋白V特异性抗体,我们观察到肌球蛋白V集中在生长锥中富含细胞器的区域。肌球蛋白V存在于一群独特的小(50-100纳米)细胞器上,以及肌动蛋白丝和质膜上。与肌球蛋白V相关的细胞器存在于微管和肌动蛋白丝上。这些结果表明,肌球蛋白V可能作为乘客搭载在沿微管运输的细胞器上,并且这些细胞器也可能能够沿肌动蛋白丝移动。此外,我们发现稀释致死小鼠的SCG生长锥在生长、形态或细胞骨架组织方面没有异常。这些结果表明,肌球蛋白V对于生长锥运动所需的牵引力、肌动蛋白细胞骨架的组织或丝状伪足动力学并非必需。