title: Long-lasting aberrant tubulovesicular membrane inclusions accumulate in developing motoneurons after a sublethal excitotoxic insult: a possible model for neuronal pathology in neurodegenerative disease.
abstract: We have previously shown that chronic treatment of chick embryos [from embryonic day 5 (E5) to E9] with NMDA rescues spinal cord motoneurons (MNs) from programmed cell death. In this situation, MNs exhibit a reduced vulnerability to acute excitotoxic lesions and downregulate NMDA and AMPA-kainate receptors. Here, we report that this treatment results in long-lasting sublethal structural changes in MNs. In Nissl-stained sections from the spinal cord of NMDA-treated embryos, MNs display an area adjacent to an eccentrically positioned nucleus in which basophilia is excluded. Ultrastructurally, MNs accumulate tubulovesicular structures surrounded by Golgi stacks. Thiamine pyrophosphatase but not acid phosphatase was detected inside the tubulovesicular structures, which are resistant to disruption by brefeldin A or monensin. Immunocytochemistry reveals changes in the content and distribution of calcitonin gene-related peptide, the KDEL receptor, the early endosomal marker EEA1, and the recycling endosome marker Rab11, indicating that a dysfunction in membrane trafficking and protein sorting occurs in these MNs. FM1-43, a marker of the endocytic pathway, strongly accumulates in MNs from isolated spinal cords after chronic NMDA treatment. Changes in the distribution of cystatin C and presenilin-1 and an accumulation of amyloid precursor protein and beta-amyloid product were also observed in NMDA-treated MNs. None of these alterations involve an interruption of MN-target (muscle) connections, as detected by the retrograde tracing of MNs with cholera toxin B subunit. These results demonstrate that chronic NMDA treatment induces severe changes in the motoneuronal endomembrane system that may be related to some neuropathological alterations described in human MN disease.
authors.affiliation: Unitat de Neurobiologia Cellular, Departament de Ciències Mèdiques Bàsiques, Facultat de Medicina, Universitat de Lleida, E25198 Lleida, Catalonia, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Tarabal
authors.forename: J
authors.initials: J
authors.lastname: Calderó
authors.forename: J
authors.initials: J
authors.lastname: Lladó
authors.forename: R W
authors.initials: RW
authors.lastname: Oppenheim
authors.forename: J E
authors.initials: JE
authors.lastname: Esquerda
mesh_terms: D000818:Animals; D020126:Brefeldin A / Q000494:pharmacology; D002451:Cell Compartmentation / Q000187:drug effects; D002642:Chick Embryo; D004195:Disease Models, Animal; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000648:ultrastructure; D011992:Endosomes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000187:drug effects / Q000648:ultrastructure; D002479:Inclusion Bodies / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D007425:Intracellular Membranes / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008565:Membrane Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D016472:Motor Neuron Disease / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D009046:Motor Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D016202:N-Methylaspartate; D009469:Neuromuscular Junction / Q000187:drug effects / Q000473:pathology; D053764:Presenilin-1; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D014617:Vacuoles / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D021601:trans-Golgi Network / Q000187:drug effects / Q000648:ultrastructure
chemical_list: D008565:Membrane Proteins; C508853:PSEN1 protein, human; D053764:Presenilin-1; D011500:Protein Synthesis Inhibitors; D020126:Brefeldin A; D016202:N-Methylaspartate