Authors

Christopher P Webster1,‡, Emma F Smith1,‡, Claudia S Bauer1, Annekathrin Moller1, Guillaume M
Hautbergue1, Laura Ferraiuolo1, Monika A Myszczynska1, Adrian Higginbottom1, Matthew J Walsh1, Alexander
J Whitworth2,†, Brian K Kaspar3, KathrinMeyer3, Pamela J Shaw1, Andrew J Grierson1,§ & Kurt J De Vos1,§,*

Affiliations

1Sheffield Institute for Translational Neuroscience (SITraN), Department of Neuroscience University of Sheffield, Sheffield, UK
2Department of Biomedical Science, University of Sheffield, Sheffield, UK
3The Research Institute at Nationwide Children's Hospital, Columbus, OH, USA
4MRC Mitochondrial Biology Unit, Cambridge, UK
↵*Corresponding author. Tel: +44 114 222 2241; E‐mail: k.de_vos@sheffield.ac.uk
↵† These authors contributed equally to this work

↵‡ These authors contributed equally to this work

Application Area

Neuroscience

General Fluorescence Microscopy

Abstract

A GGGGCC hexanucleotide repeat expansion in the C9orf72 gene is the most common genetic cause of amyotrophic lateral sclerosis and frontotemporal dementia (C9ALS/FTD). C9orf72 encodes two C9orf72 protein isoforms of unclear function. Reduced levels of C9orf72 expression have been reported in C9ALS/FTD patients, and although C9orf72 haploinsufficiency has been proposed to contribute to C9ALS/FTD, its significance is not yet clear. Here, we report that C9orf72 interacts with Rab1a and the Unc-51-like kinase 1 (ULK1) autophagy initiation complex. As a Rab1a effector, C9orf72 controls initiation of autophagy by regulating the Rab1a-dependent trafficking of the ULK1 autophagy initiation complex to the phagophore. Accordingly, reduction of C9orf72 expression in cell lines and primary neurons attenuated autophagy and caused accumulation of p62-positive puncta reminiscent of the p62 pathology observed in C9ALS/FTD patients. Finally, basal levels of autophagy were markedly reduced in C9ALS/FTD patient-derived iNeurons. Thus, our data identify C9orf72 as a novel Rab1a effector in the regulation of autophagy and indicate that C9orf72 haploinsufficiency and associated reductions in autophagy might be the underlying cause of C9ALS/FTD-associated p62 pathology.

Extract

Images were recorded using appropriate filtersets (Omega Optical and Chroma Technology) using MicroManager 1.4 software (Edelstein et al, 2014) on a Zeiss Axioplan2 microscope fitted with a Hamamatsu C4880‐80 or Retiga R3 (QImaging) CCD camera, PE‐300 LED illumination (CoolLED)

Product Associated Features

CoolLED’s pE-300white offers intense, broad-spectrum LED illumination for imaging most common fluorescent stains.

Diascopic Technique

-

Live Cell Issues

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Product Type

pE-300white, pE-4000

Journal

-

Year of Publication

2016

Country of Publication

UK