Redmann Veronika, Lamb Christopher A, Hwang Seungmin, Orchard Robert C, Kim Sungsu, Razi Minoo, Milam Ashley, Park Sunmin, Yokoyama Christine C, Kambal Amal, Kreamalmeyer Darren, Bosch Marie K, Xiao Maolei, Green Karen, Kim Jungsu, Pruett-Miller Shondra M, Ornitz David M, Allen Paul M, Beatty Wandy L, Schmidt Robert E, DiAntonio Aaron, Tooze Sharon A, Virgin Herbert W
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
The Francis Crick Institute, Lincoln's Inn Fields Laboratory, London, WC2A 3LY, UK.
Sci Rep. 2016 Mar 18;6:23326. doi: 10.1038/srep23326.
CLEC16A is in a locus genetically linked to autoimmune diseases including multiple sclerosis, but the function of this gene in the nervous system is unknown. Here we show that two mouse strains carrying independent Clec16a mutations developed neurodegenerative disease characterized by motor impairments and loss of Purkinje cells. Neurons from Clec16a-mutant mice exhibited increased expression of the autophagy substrate p62, accumulation of abnormal intra-axonal membranous structures bearing the autophagy protein LC3, and abnormal Golgi morphology. Multiple aspects of endocytosis, lysosome and Golgi function were normal in Clec16a-deficient murine embryonic fibroblasts and HeLa cells. However, these cells displayed abnormal bulk autophagy despite unimpaired autophagosome formation. Cultured Clec16a-deficient cells exhibited a striking accumulation of LC3 and LAMP-1 positive autolysosomes containing undigested cytoplasmic contents. Therefore Clec16a, an autophagy protein that is critical for autolysosome function and clearance, is required for Purkinje cell survival.
CLEC16A位于一个与包括多发性硬化症在内的自身免疫性疾病存在遗传关联的基因座中,但该基因在神经系统中的功能尚不清楚。在此我们表明,携带独立Clec16a突变的两种小鼠品系发生了以运动障碍和浦肯野细胞丢失为特征的神经退行性疾病。来自Clec16a突变小鼠的神经元表现出自噬底物p62表达增加、带有自噬蛋白LC3的轴突内异常膜性结构积累以及高尔基体形态异常。在Clec16a缺陷的小鼠胚胎成纤维细胞和HeLa细胞中,内吞作用、溶酶体和高尔基体功能的多个方面均正常。然而,尽管自噬体形成未受损,但这些细胞表现出异常的整体自噬。培养的Clec16a缺陷细胞表现出LC3和LAMP - 1阳性自噬溶酶体的显著积累,其中含有未消化的细胞质内容物。因此,Clec16a作为一种对自噬溶酶体功能和清除至关重要的自噬蛋白,是浦肯野细胞存活所必需的。