title: Hereditary spastic paraplegia-associated mutations in the NIPA1 gene and its Caenorhabditis elegans homolog trigger neural degeneration in vitro and in vivo through a gain-of-function mechanism.
abstract: We studied the consequences of expression of wild-type (WT) human NIPA1 and two mutant forms of NIPA1 with known HSP-associated mutations (T45R and G106R) on cultured rat cortical neurons and using equivalent substitutions in the Caenorhabditis elegans NIPA1 homolog CeNIPA. WT NIPA1 localized in transfected neuronal and non-neuronal cells to the Golgi complex, a subset of synaptic vesicles, to a subset of early endosomes, and plasma cell membrane. Mutant NIPA1 accumulated in the endoplasmic reticulum (ER) triggering ER stress and features of apoptotic cell death. Flow cytometric analysis of NIPA1 surface expression demonstrated relatively intact trafficking of mutant forms and only the T45R mutant exhibited modestly reduced patterns of surface expression without evidence for a dominant-negative effect. In vivo pan-neuronal expression of the WT C. elegans NIPA1 homolog (CeNIPA) was well tolerated, with no obvious impact on neuronal morphology or behavior. In striking contrast, expression of CeNIPA bearing HSP-associated mutations caused a progressive neural degeneration and a clear motor phenotype. Neuronal loss in these animals began at day 7 and by day 9 animals were completely paralyzed. These effects appeared to arise from activation of the apoptotic program triggered by unfolded protein response (UPR), as we observed marked modifications of motor and cellular phenotype when mutant NIPA1 was expressed in caspase (ced-3)- and UPR (xbp-1)-deficient backgrounds. We propose that HSP-associated mutations in NIPA1 lead to cellular and functional deficits through a gain-of-function mechanism supporting the ER accumulation of toxic NIPA1 proteins.
authors.affiliation: Department of Neurology, Vanderbilt University, Nashville, Tennessee 37232-8552, USA.
authors.forename: Jiali
authors.initials: J
authors.lastname: Zhao
authors.forename: Dawn S
authors.initials: DS
authors.lastname: Matthies
authors.forename: Emmanuel J
authors.initials: EJ
authors.lastname: Botzolakis
authors.forename: Robert L
authors.initials: RL
authors.lastname: Macdonald
authors.forename: Randy D
authors.initials: RD
authors.lastname: Blakely
authors.forename: Peter
authors.initials: P
authors.lastname: Hedera
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001370:Axonal Transport / Q000235:genetics; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004195:Disease Models, Animal; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D009154:Mutation; D009410:Nerve Degeneration / Q000235:genetics* / Q000473:pathology; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D009928:Organ Specificity; D010243:Paralysis / Q000235:genetics; D010641:Phenotype; D021381:Protein Transport / Q000235:genetics; D051381:Rats; D015419:Spastic Paraplegia, Hereditary / Q000235:genetics*; D013572:Synaptic Vesicles / Q000378:metabolism; D014162:Transfection
chemical_list: D029742:Caenorhabditis elegans Proteins; D008565:Membrane Proteins; C478824:NIPA1 protein, human