Glial cells maintain synapses by inhibiting an activity-dependent retrograde protease signal
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ABSTRACT:
PROVIDER: PRJNA516573 | ENA |
REPOSITORIES: ENA
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Project description:We utilized genetic methods to examine how Schwann cells prevent degeneration of motor neurons (MNs) in the spinal cord. Blocking peripheral, neuromuscular activity completely rescued MNs and neuromuscular junctions (NMJs) in erbB3 mutant mice lacking Schwann cells, which normally exhibit profound neurodegeneration. We searched for the molecular basis of this effect by examining the transcriptomes (all of the expressed genes) in the muscle of control mice with Schwann cells and erbB3 mutant mice without them. We found evidence that a negative signal expressed by muscle was regulated by neural activity and normally blocked by factors produced in Schwann cells. When we eliminated this activity-induced negative signal (thrombin) from muscle, MNs and NMJs were protected in erbB3 mutants, similar to the effects of eliminating activity. Together, these results suggest that Schwann cells prevent neurodegeneration by inhibiting the effect of activity-induced, muscle-derived negative factors, rather than by providing trophic positive factors.
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Project description:Bergmann glial cells of the vertebrate cerebellum play essential roles in the development and maintenance of cerebellar structure and function. During development, Bergmann glia provide structural support to the expanding cerebellar anlage and also serve as guides for migrating neurons (granule cells). As the cerebellum matures, Bergmann glia become important in dendritic arborization, synapse maintenance and synaptic function. The molecular mechanisms underlying these diverse and important functions of Bergmann glia remain largely unknown. We used microarray analysis to examine global gene expression in individual Bergmann glial cells derived at P6 (a time of extensive neuronal migration and cerebellar growth) and at P30 (when cerebellar development is complete and Bergmann glia play important roles at synapses). After gentle dissociation of cerebellar tissue derived from mice expressing GFP under the GFAP gene promoter (GFAP-GFP mice) (Zhuo, L., et al., 1997, Developmental biology), single GFP-positive Bergmann glial cells were aspirated into microcapillary tubes. Amplified cDNAs were prepared from single cells using RT-PCR and hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 expression arrays (one array per cell). Five P6 cells and five P30 cells were used to generate the data presented in this study.
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Project description:Bergmann glial cells of the vertebrate cerebellum play essential roles in the development and maintenance of cerebellar structure and function. During development, Bergmann glia provide structural support to the expanding cerebellar anlage and also serve as guides for migrating neurons (granule cells). As the cerebellum matures, Bergmann glia become important in dendritic arborization, synapse maintenance and synaptic function. The molecular mechanisms underlying these diverse and important functions of Bergmann glia remain largely unknown. We used microarray analysis to examine global gene expression in individual Bergmann glial cells derived at P6 (a time of extensive neuronal migration and cerebellar growth) and at P30 (when cerebellar development is complete and Bergmann glia play important roles at synapses).
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Project description:Astrocytes are increasingly recognized as crucial contributors to neuronal function at synapses, axons, and somas. Reliable methods that can provide insight into the astrocyte proteins at the neuron-astrocyte functional interface are highly desirable. Here we conducted a mass spectrometry analysis of Percoll® gradient isolated gliosomes, a viable preparation of glial subcellular particles often used to study mechanisms of astrocytic transmitter uptake and release and their regulation. Gliosomes were compared with synaptosomes, a preparation containing the neurotransmitter release machinery, and accordingly synaptosomes were enriched for proteins involved in synaptic vesicle mediated transport. Interestingly, gliosome preparations were found to be enriched for different classes of known astrocyte proteins, such as VAMP3 (involved in astrocyte exocytosis), Ezrin (perisynaptic astrocyte cytoskeletal protein), and Basigin (astrocyte membrane glycoprotein), as well as for G-protein mediated signaling proteins. Together, these data provide the first detailed description of the gliosome proteome and show that gliosomes can be a useful preparation to study glial membrane proteins and associated processes.
2014-11-17 | PXD001375 | Pride
Project description:Muscle denervation due to injury, disease or aging results in impaired motor function. Restoring neuromuscular communication requires axonal regrowth and regeneration of neuromuscular synapses. Muscle activity inhibits neuromuscular synapse regeneration. The mechanism by which muscle activity regulates regeneration of synapses is poorly understood. Dach2 and Hdac9 are activity-regulated transcriptional co-repressors that are highly expressed in innervated muscle and suppressed following muscle denervation. Here, we report that Dach2 and Hdac9 inhibit regeneration of neuromuscular synapses. Importantly, we identified Myog and Gdf5 as muscle-specific Dach2/Hdac9-regulated genes that stimulate neuromuscular regeneration in denervated muscle. Interestingly, Gdf5 also stimulates presynaptic differentiation and inhibits branching of regenerating neurons. Finally, we found that Dach2 and Hdac9 suppress miR206 expression, a microRNA involved in enhancing neuromuscular regeneration. RNAseq on innervated and 3 day denervated adult soleus muscle from wildtype mice is compared with that from 3 day denervated soleus muscle from Dach2/Hdac9 deleted mice to identify Dach2/Hdac9-regulated genes.
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