Regulation of endosomal motility and degradation by amyotrophic lateral sclerosis 2/alsin

Lai Chen, Xie Chengsong, Shim Hoon, Chandran Jayanth, Howell, Cai, Cai Huaibin

Description

Dysfunction of alsin, particularly its putative Rab5 guanine-nucleotide-exchange factor activity, has been linked to one form of juvenile onset recessive familial amyotrophic lateral sclerosis (ALS2). Multiple lines of alsin knockout ( ALS2 -/- ) mice have been generated to model this disease. However, it remains elusive whether the Rab5-dependent endocytosis is altered in ALS2 -/- neurons. To directly examine the Rab5-mediated endosomal trafficking in ALS2 -/- neurons, we introduced green fluorescent protein (GFP)-tagged Rab5 into cultured hippocampal neurons to monitor the morphology and motility of Rab5-associated early endosomes. Here we report that Rab5-mediated endocytosis was severely altered in ALS2 -/- neurons. Excessive accumulation of Rab5-positive vesicles was observed in ALS2 -/- neurons, which correlated with a significant reduction in endosomal motility and augmentation in endosomal conversion to lysosomes. Consequently, a significant increase in endosome/lysosome-dependent degradation of internalized glutamate receptors was observed in ALS2 -/- neurons. These phenotypes closely resembled the endosomal trafficking abnormalities induced by a constitutively active form of Rab5 in wild-type neurons. Therefore, our findings reveal a negatively regulatory mechanism of alsin in Rab5-mediated endosomal trafficking, suggesting that enhanced endosomal degradation in ALS2 -/- neurons may underlie the pathogenesis of motor neuron degeneration in ALS2 and related motor neuron diseases.

Informations

• Publié par: biomed
• Publié le: 01 janvier 2009
• Nombre de lectures: 27
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Molecular Brain

BioMedCentral

Open Access Research Regulation of endosomal motility and degradation by amyotrophic lateral sclerosis 2/alsin 1 11,3 1,4 Chen Lai*, Chengsong Xie, Hoon Shim, Jayanth Chandran, 2 1 Brian W Howelland Huaibin Cai*

1 2 Address: Laboratoryof Neurogenetics, National Institute on Aging, National Institutes of Health, Bethesda, MD 20892 USA,Neurogenetics 3 Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892 USA,Current address: School 4 of Medicine at Virginia Commonwealth University, Richmond, VA 23298, USA andCurrent address: University of Edinburgh, Centre for Inflammation Research, The Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh, EH16 4TJ, UK Email: Chen Lai*  [e-mail]; Chengsong Xie  [e-mail]; Hoon Shim  [e-mail]; Jayanth Chandran  [e-mail]; Brian W Howell  [e-mail]; Huaibin Cai*  [e-mail] * Corresponding authors

Published: 24 July 2009Received: 30 June 2009 Accepted: 24 July 2009 Molecular Brain2009,2:23 doi:10.1186/1756-6606-2-23 This article is available from: http://www.molecularbrain.com/content/2/1/23 © 2009 Lai et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract Dysfunction of alsin, particularly its putative Rab5 guanine-nucleotide-exchange factor activity, has been linked to one form of juvenile onset recessive familial amyotrophic lateral sclerosis (ALS2). -/- Multiple lines of alsin knockout (ALS2) mice have been generated to model this disease. However, -/- it remains elusive whether the Rab5-dependent endocytosis is altered inALS2neurons. To -/- directly examine the Rab5-mediated endosomal trafficking inALS2neurons, we introduced green fluorescent protein (GFP)-tagged Rab5 into cultured hippocampal neurons to monitor the morphology and motility of Rab5-associated early endosomes. Here we report that Rab5-mediated -/- endocytosis was severely altered inALS2neurons. Excessive accumulation of Rab5-positive -/- vesicles was observed inALS2neurons, which correlated with a significant reduction in endosomal motility and augmentation in endosomal conversion to lysosomes. Consequently, a significant increase in endosome/lysosome-dependent degradation of internalized glutamate receptors was -/- observed inALS2neurons. These phenotypes closely resembled the endosomal trafficking abnormalities induced by a constitutively active form of Rab5 in wild-type neurons. Therefore, our findings reveal a negatively regulatory mechanism of alsin in Rab5-mediated endosomal trafficking, -/- suggesting that enhanced endosomal degradation inALS2neurons may underlie the pathogenesis of motor neuron degeneration in ALS2 and related motor neuron diseases.

Background Amyotrophic Lateral Sclerosis (ALS) is a neurodegenera tive disease caused by the selective degeneration of spinal and corticospinal motor neurons, resulting in muscle weakness and atrophy along with spastic paralysis [1,2]. One form of inherited juvenileonset amyotrophic lateral sclerosis (ALS2) is caused by loss of function mutations in theALS2gene [36]. Elucidation of the function(s) of

alsin is essential in [7]understanding the pathogenic mechanism of this type of motor neuron disease.

Alsin, encoded by the fulllengthALS2gene, contains three putative guaninenucleotideexchange factor (GEF) domains based on the sequence homology [4,6]. Previous studies indicate that the carboxylterminal vacuolar pro tein sorting 9 (VPS9)like domain in conjunction with the

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