title: Trophic support delays but does not prevent cell-intrinsic degeneration of neurons deficient for munc18-1.
abstract: The stability of neuronal networks is thought to depend on synaptic transmission which provides activity-dependent maintenance signals for both synapses and neurons. Here, we tested the relationship between presynaptic secretion and neuronal maintenance using munc18-1-null mutant mice as a model. These mutants have a specific defect in secretion from synaptic and large dense-cored vesicles [Verhage et al. (2000), Science, 287, 864-869; Voets et al. (2001), Neuron, 31, 581-591]. Neuronal networks in these mutants develop normally up to synapse formation but eventually degenerate. The proposed relationship between secretion and neuronal maintenance was tested in low-density and organotypic cultures and, in vivo, by conditional cell-specific inactivation of the munc18-1 gene. Dissociated munc18-1-deficient neurons died within 4 days in vitro (DIV). Application of trophic factors, insulin or BDNF delayed degeneration up to 7 DIV. In organotypic cultures, munc18-1-deficient neurons survived until 9 DIV. On glial feeders, these neurons survived up to 10 DIV and 14 DIV when insulin was applied. Co-culturing dissociated mutant neurons with wild-type neurons did not prolong survival beyond 4 DIV, but coculturing mutant slices with wild-type slices prolonged survival up to 19 DIV. Cell-specific deletion of munc18-1 expression in cerebellar Purkinje cells in vivo resulted in the specific loss of these neurons without affecting connected or surrounding neurons. Together, these data allow three conclusions. First, the lack of synaptic activity cannot explain the degeneration in munc18-1-null mutants. Second, trophic support delays but cannot prevent degeneration. Third, a cell-intrinsic yet unknown function of munc18-1 is essential for prolonged survival.
authors.affiliation: Department of Molecular Neuroscience, Rudolf Magnus Institute, University of Utrecht Medical Center, Utrecht, The Netherlands.
authors.forename: Joost H
authors.initials: JH
authors.lastname: Heeroma
authors.forename: Martijn
authors.initials: M
authors.lastname: Roelandse
authors.forename: Keimpe
authors.initials: K
authors.lastname: Wierda
authors.forename: Karlijn I
authors.initials: KI
authors.lastname: van Aerde
authors.forename: Ruud F G
authors.initials: RF
authors.lastname: Toonen
authors.forename: Robert A
authors.initials: RA
authors.lastname: Hensbroek
authors.forename: Arjen
authors.initials: A
authors.lastname: Brussaard
authors.forename: Andrew
authors.initials: A
authors.lastname: Matus
authors.forename: Matthijs
authors.initials: M
authors.lastname: Verhage
mesh_terms: D000200:Action Potentials / Q000235:genetics / Q000502:physiology / Q000528:radiation effects; D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000627:therapeutic use; D064026:Calbindins; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D004558:Electric Stimulation / Q000379:methods; D004622:Embryo, Mammalian; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005968:Glutamate Decarboxylase / Q000378:metabolism; D006624:Hippocampus / Q000473:pathology* / Q000503:physiopathology; D007150:Immunohistochemistry / Q000379:methods; D007328:Insulin / Q000627:therapeutic use; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D051938:Munc18 Proteins; D009410:Nerve Degeneration / Q000188:drug therapy / Q000235:genetics / Q000503:physiopathology*; D009414:Nerve Growth Factors / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018377:Neurotransmitter Agents / Q000235:genetics / Q000378:metabolism; D018408:Patch-Clamp Techniques / Q000379:methods; D010640:Phenothiazines; D050765:Qa-SNARE Proteins; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000378:metabolism; D013569:Synapses / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D033921:Vesicular Transport Proteins / Q000172:deficiency* / Q000235:genetics / Q000502:physiology
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D064026:Calbindins; D005904:Glial Fibrillary Acidic Protein; D007328:Insulin; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D051938:Munc18 Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; D010640:Phenothiazines; D050765:Qa-SNARE Proteins; D064030:S100 Calcium Binding Protein G; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; C497938:Stxbp1 protein, mouse; D033921:Vesicular Transport Proteins; D005968:Glutamate Decarboxylase; C009469:thionine