Autophagy is a prosurvival mechanism in cells expressing an autosomal dominant familial neurohypophyseal diabetes insipidus mutant vasopressin transgene.
Autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI) is a progressive, inherited neurodegenerative disorder that presents as polydipsia and polyuria as a consequence of a loss of secretion of the antidiuretic hormone vasopressin (VP) from posterior pituitary nerve terminals. VP gene mutations cause adFNDI. Rats expressing an adFNDI VP transgene (Cys67stop) show a neuronal pathology characterized by autophagic structures in the cell body. adFNDI has thus been added to the list of protein aggregation diseases, along with Alzheimer's, Parkinson's and Huntington's, which are associated with autophagy, a bulk process that delivers regions of cytosol to lysosomes for degradation. However, the role of autophagy in these diseases is unclear. To address the relationships between mutant protein accumulation, autophagy, cell survival, and cell death, we have developed a novel and tractable in vitro system. We have constructed adenoviral vectors (Ads) that express structural genes encoding either the Cys67stop mutant protein (Ad-VCAT-Cys67stop) or an epitope-tagged wild-type VP precursor (Ad-VCAT). After infection of mouse neuroblastoma Neuro2a cells, Ad-VCAT encoded material enters neurite processes and accumulates in terminals, while the Cys67stop protein is confined to enlarged vesicles in the cell body. Similar to the intracellular derangements seen in the Cys67stop rats, these structures are of ER origin, and colocalize with markers of autophagy. Neither Ad-VCAT-Cys67stop nor Ad-VCAT expression affected cell viability. However, inhibition of autophagy or lysosomal protein degradation, while having no effect on Ad-VCAT-expressing cells, significantly increased apoptotic cell death following Ad-VCAT-Cys67stop expression. These data suggest that activation of autophagy by the stress of the expression of an adFNDI mutant protein is a prosurvival mechanism.
Castino
Roberta
R
Molecular Neuroendocrinology Research Group, Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, University of Bristol, England, UK.
Davies
Janet
J
Beaucourt
Stephanie
S
Isidoro
Ciro
C
Murphy
David
D
D000165:Acridine Orange; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified; D001343:Autophagy / Q000235:genetics*; D015153:Blotting, Western; D002103:Cadaverine / Q000031:analogs & derivatives; D002402:Cathepsin D / Q000032:analysis; D045744:Cell Line, Tumor; D018388:Codon, Terminator / Q000235:genetics; D003545:Cysteine / Q000235:genetics; D020790:Diabetes Insipidus, Neurogenic / Q000235:genetics* / Q000473:pathology; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D015870:Gene Expression; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000378:metabolism; D009154:Mutation; D016501:Neurites / Q000378:metabolism; D009447:Neuroblastoma; D009474:Neurons / Q000473:pathology; D015388:Organelles; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D019076:Transgenes / Q000235:genetics*; D014617:Vacuoles; D014667:Vasopressins / Q000235:genetics* / Q000378:metabolism
D018388:Codon, Terminator; D005456:Fluorescent Dyes; D011994:Recombinant Proteins; D014667:Vasopressins; D002402:Cathepsin D; D000165:Acridine Orange; C008542:monodansylcadaverine; D003545:Cysteine; D002103:Cadaverine