TY - JOUR
T1 - Early golgi abnormalities and neurodegeneration upon loss of presynaptic proteins munc18-1, syntaxin-1, or SNAP-25
AU - Santos, Tatiana C.
AU - Wierda, Keimpe
AU - Broeke, Jurjen H.
AU - Toonen, Ruud F.
AU - Verhage, Matthijs
PY - 2017/4/26
Y1 - 2017/4/26
N2 - The loss of presynaptic proteins Munc18-1, syntaxin-1, or SNAP-25 is known to produce cell death, but the underlying features have not been compared experimentally. Here, we investigated these features in cultured mouse CNS andDRGneurons. Side-by-side comparisons confirmed massive cell death, before synaptogenesis, within 1-4 DIV upon loss of t-SNAREs (syntaxin-1, SNAP-25) or Munc18-1, but not v-SNAREs (synaptobrevins/VAMP1/2/3 using tetanus neurotoxin (TeNT), also in TI-VAMP/VAMP7 knock-out (KO) neurons). A condensed cis-Golgi was the first abnormality observed upon Munc18-1 or SNAP-25 loss within 3 DIV. This phenotype was distinct from the Golgi fragmentation observed in apoptosis. Cell death was too rapid after syntaxin-1 loss to study Golgi abnormalities. Syntaxin-1 and Munc18-1 depend on each other for normal cellular levels. We observed that endogenous syntaxin-1 accumulates at the Golgi of Munc18-1 KO neurons. However, expression of a non-neuronal Munc18 isoform that does not bind syntaxin-1, Munc18-3, in Munc18-1 KO neurons prevented cell death and restored normal cis-Golgi morphology, but not synaptic transmission or syntaxin-1 targeting. Finally, we observed that DRG neurons are the only Munc18-1 KO neurons that do not degenerate in vivo or in vitro. In these neurons, cis-Golgi abnormalities were less severe, with no changes in Golgi shape. Together, these data demonstrate that cell death upon Munc18-1, syntaxin-1, or SNAP-25 loss occurs via a degenerative pathway unrelated to the known synapse function of these proteins and involving early cis-Golgi abnormalities, distinct from apoptosis.
AB - The loss of presynaptic proteins Munc18-1, syntaxin-1, or SNAP-25 is known to produce cell death, but the underlying features have not been compared experimentally. Here, we investigated these features in cultured mouse CNS andDRGneurons. Side-by-side comparisons confirmed massive cell death, before synaptogenesis, within 1-4 DIV upon loss of t-SNAREs (syntaxin-1, SNAP-25) or Munc18-1, but not v-SNAREs (synaptobrevins/VAMP1/2/3 using tetanus neurotoxin (TeNT), also in TI-VAMP/VAMP7 knock-out (KO) neurons). A condensed cis-Golgi was the first abnormality observed upon Munc18-1 or SNAP-25 loss within 3 DIV. This phenotype was distinct from the Golgi fragmentation observed in apoptosis. Cell death was too rapid after syntaxin-1 loss to study Golgi abnormalities. Syntaxin-1 and Munc18-1 depend on each other for normal cellular levels. We observed that endogenous syntaxin-1 accumulates at the Golgi of Munc18-1 KO neurons. However, expression of a non-neuronal Munc18 isoform that does not bind syntaxin-1, Munc18-3, in Munc18-1 KO neurons prevented cell death and restored normal cis-Golgi morphology, but not synaptic transmission or syntaxin-1 targeting. Finally, we observed that DRG neurons are the only Munc18-1 KO neurons that do not degenerate in vivo or in vitro. In these neurons, cis-Golgi abnormalities were less severe, with no changes in Golgi shape. Together, these data demonstrate that cell death upon Munc18-1, syntaxin-1, or SNAP-25 loss occurs via a degenerative pathway unrelated to the known synapse function of these proteins and involving early cis-Golgi abnormalities, distinct from apoptosis.
KW - Animals
KW - Apoptosis
KW - Cell Death
KW - Cells, Cultured
KW - Exocytosis
KW - Golgi Apparatus
KW - Journal Article
KW - Mice
KW - Mice, Knockout
KW - Munc18 Proteins
KW - Neurodegeneration
KW - Neurodegenerative Diseases
KW - Presynaptic proteins
KW - Synapses
KW - Synaptic transmission
KW - Synaptosomal-Associated Protein 25
KW - Syntaxin 1
UR - http://www.scopus.com/inward/record.url?scp=85019095167&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85019095167&partnerID=8YFLogxK
U2 - https://doi.org/10.1523/JNEUROSCI.3352-16.2017
DO - https://doi.org/10.1523/JNEUROSCI.3352-16.2017
M3 - Article
C2 - 28348137
SN - 0270-6474
VL - 37
SP - 4525
EP - 4539
JO - Journal of neuroscience
JF - Journal of neuroscience
IS - 17
ER -