title: Protein product of CLN6 gene responsible for variant late-onset infantile neuronal ceroid lipofuscinosis interacts with CRMP-2.
abstract: Mutations in CLN6 cause variant late-onset neuronal ceroid lipofuscinosis (vLINCL), a childhood neurodegenerative disorder resulting from aberrant neuronal cell loss and pathological accumulation of lysosomal autofluorescent storage material in the central nervous system. The direct function of the endoplasmic reticulum-resident protein CLN6 and how dysfunction of this protein results in vLINCL are unknown. We report that CLN6 interacts with collapsin response mediator protein-2 (CRMP-2). To further understand the significance and possible contribution to vLINCL of the CLN6-CRMP-2 interaction, we utilized the nclf mouse, which harbors mutations in CLN6. Significantly, CRMP-2 protein level was found to be reduced in the nclf mouse brain, particularly in the thalamus. Because CRMP-2 functions in growth cone collapse and is an effector protein downstream of Sema3A signaling, this pathway was examined via a dorsal root ganglion (DRG) repulsion assay. However, there were no defects in the repulsion of DRGs derived from nclf mice, indicating that the loss of CLN6 does not affect Sema3A signaling. CRMP-2 has also been implicated in controlling axon number and outgrowth, as observed in cultured hippocampal neurons. Therefore, we explored the formation and maturation of hippocampal neurons derived from nclf mice in a glial coculture system. The maturation of these neurons was reduced; by day in vitro (DIV) 8, more than 50% of nclf-derived hippocampal neurons had died. Additionally, beginning around DIV4, nclf neurons were less mature than their WT counterparts, presumably because of an inability to form mature synaptic connections. We concluded that alterations in neurite maturation resulting from a loss of CLN6-CRMP-2 interaction may contribute to neuronal dysfunction and pathology in vLINCL.
authors.affiliation: Center for Neural Development and Disease, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.
authors.forename: Jared W
authors.initials: JW
authors.lastname: Benedict
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Getty
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Wishart
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Gillingwater
authors.forename: David A
authors.initials: DA
authors.lastname: Pearce
mesh_terms: D000818:Animals; D001921:Brain / Q000254:growth & development / Q000378:metabolism / Q000503:physiopathology; D002454:Cell Differentiation / Q000235:genetics; D018920:Coculture Techniques; D005727:Ganglia, Spinal / Q000378:metabolism / Q000473:pathology; D020439:Growth Cones / Q000378:metabolism / Q000473:pathology; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D041681:NIH 3T3 Cells; D009410:Nerve Degeneration / Q000235:genetics* / Q000378:metabolism* / Q000503:physiopathology; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009434:Neural Pathways / Q000254:growth & development / Q000378:metabolism / Q000503:physiopathology; D055495:Neurogenesis / Q000235:genetics; D009472:Neuronal Ceroid-Lipofuscinoses / Q000235:genetics* / Q000378:metabolism* / Q000503:physiopathology; D013569:Synapses / Q000378:metabolism / Q000473:pathology
chemical_list: C486722:Cln6 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C095114:collapsin response mediator protein-2