ABSTRACT
Synaptic elements from neuromuscular junctions (NMJs) undergo massive morphological and functional changes upon nerve injury. While morphological changes of NMJ-associated glia in response to injury has been investigated, their functional properties remain elusive. Perisynaptic Schwann cells (PSCs), glial cells at the NMJ, are essential for NMJ maintenance and repair, and are involved in synaptic efficacy and plasticity. Importantly, these functions are regulated by PSCs ability to detect synaptic transmission through, notably, muscarinic (mAChRs) and purinergic receptors' activation. Using Ca2+ imaging and electrophysiological recordings of synaptic transmission at the mouse NMJ, we investigated PSC receptors activation following denervation and during reinnervation in adults and at denervated NMJs in an ALS mouse model (SOD1G37R ). We observed reduced PSCs mAChR-mediated Ca2+ responses at denervated and reinnervating NMJs. Importantly, PSC phenotypes during denervation and reinnervation were distinct than the one observed during NMJ maturation. At denervated NMJs, exogenous activation of mAChRs greatly diminished galectin-3 expression, a glial marker of phagocytosis. PSCs Ca2+ responses at reinnervating NMJs did not correlate with the number of innervating axons or process extensions. Interestingly, we observed an extended period of reduced PSC mAChRs activation after the injury (up to 60 days), suggesting a glial memory of injury. PSCs associated with denervated NMJs in an ALS model (SOD1G37R mice) did not show any muscarinic adaptation, a phenotype incompatible with NMJ repair. Understanding functional mechanisms that underlie this glial response to injury may contribute to favor complete NMJ and motor recovery.
Subject(s)
Amyotrophic Lateral Sclerosis , Amyotrophic Lateral Sclerosis/metabolism , Animals , Cholinergic Agents/metabolism , Mice , Neuroglia/physiology , Neuromuscular Junction/metabolism , Schwann Cells/metabolism , Superoxide Dismutase-1/metabolismABSTRACT
The formation of highly efficient and reliable synapses at the neuromuscular junction (NMJ) relies on dynamic molecular interactions. Studies of the development and maturation of the NMJ have focused on events that are dependent on synaptic activity and that require the coordinated actions of nerve- and muscle-derived molecules with different targets and effects. More recently, perisynaptic Schwann cells--the glial cells at NMJs--have become an important focus of research. These glia concomitantly contribute to pre- and postsynaptic maturation while undergoing maturation themselves. Thus, an intricate 'danse à trois''regulates the maturation of the NMJ to form a highly efficient communication unit, in which fine glial processes lie in close proximity to a highly concentrated population of postsynaptic receptors and perfectly aligned presynaptic release sites.
Subject(s)
Neurogenesis/physiology , Neuromuscular Junction/growth & development , Synaptic Transmission/physiology , Animals , Neuroglia/physiology , Neuromuscular Junction/metabolism , Signal Transduction/physiologyABSTRACT
Secreted Protein Acidic and Rich in Cysteine (SPARC) is a matricellular protein produced by glial cells. Although it is highly expressed in synaptogenic areas in the developing nervous system, it is still unclear whether this molecule displays an action on synaptic activity. We show that nanomolar concentrations of SPARC favour a more efficient synapse formation and increase short term depression in single cell cholinergic microcultures. The change in synaptic plasticity, which is also observed when SPARC is locally secreted on stable synapses for 24-48 h, is caused by a high release probability and a reduction in the size of the rapidly releasable pool of vesicles. Both features are attributable to synapses operating at an immature stage as demonstrated by correlative electrophysiology and electron microscopy experiments. Presynaptic terminals developed in the presence of SPARC display few cytoplasmic vesicles and two to threefold decrease in the number of docked vesicles at active zones. At the postsynaptic level, the analysis of miniature excitatory postsynaptic currents suggests SPARC has little effect on the number of nicotinic receptors but might alter their composition. The widespread distribution of SPARC makes current findings potentially relevant to other excitatory synapses and development of neuronal circuits.
Subject(s)
Cholinergic Fibers/metabolism , Neurons/metabolism , Neurons/ultrastructure , Osteonectin/metabolism , Presynaptic Terminals/metabolism , Presynaptic Terminals/ultrastructure , Animals , Electrophysiology , Excitatory Postsynaptic Potentials/physiology , Microscopy, Electron , Neuroglia/metabolism , Neuronal Plasticity/physiology , Rats , Rats, Sprague-Dawley , Synapses/metabolism , Synapses/ultrastructure , Synaptic Transmission/physiology , Synaptic Vesicles/metabolism , Synaptic Vesicles/ultrastructureABSTRACT
Nicotinic synapses in the autonomous nervous system display use-dependent plasticity but the contribution of cellular environment, as well as the presynaptic mechanisms implicated in this process remain to be determined. To address these questions synaptic function was assayed in rat superior cervical ganglion (SCG) neurons microcultured in isolation from any other cell type and compared to those microcultured in the presence of Schwann cells of ganglionar origin. Schwann cells were not required for synapse formation in vitro because functional cholinergic autaptic synapses were established in both experimental conditions. The number of synapses was comparable between the two culture conditions but the frequency of spontaneous miniature excitatory postsynaptic currents was enhanced in those neurons grown in direct contact with glial cells. Autapses displayed facilitation and depression, both processes being determined by the fraction of vesicles from the readily releasable pool discharged by an action potential. At high release probabilities vesicles were more efficiently mobilized, thus promoting depression, whilst low release probabilities made facilitation likely to occur. Schwann cells did not modify significantly facilitation but increased synaptic depression. In single cell microcultures, paired pulse stimuli showed a monoexponential recovery from depression with a time constant of approximately 60 ms, while in microcultures developed together with glial cells, recovery was bi-exponential with a significantly slower time course. Altogether these results show that Schwann cells from sympathetic ganglia directly modulate use-dependent plasticity of nicotinic synapses in vitro by enhancing short-term depression.
Subject(s)
Cholinergic Fibers/physiology , Neuronal Plasticity , Schwann Cells/physiology , Synaptic Potentials , Animals , Cells, Cultured , Rats , Rats, Sprague-Dawley , Superior Cervical Ganglion/cytologyABSTRACT
The transcription factor CCAAT/enhancer binding protein beta (C/EBPbeta) regulates the expression of key genes in inflammation but little is known about the involvement of C/EBPbeta in glial activation. In this report, we have studied the patterns of astroglial and microglial C/EBPbeta expression in primary mouse cortical cultures. We show that both astrocytes and microglia express C/EBPbeta in untreated mixed glial cultures. C/EBPbeta is upregulated when glial activation is induced by lipopolysaccharide (LPS). The LPS-induced upregulation of glial C/EBPbeta is rapid (2 h at mRNA level, 4 h at protein level). It is elicited by low concentrations of LPS (almost maximal effect at 1 ng/mL) and it is reversed by the protein synthesis inhibitor cycloheximide. C/EBPbeta nuclear levels increase both in astrocytes and microglia after LPS treatment, and the response is more marked in microglia. The LPS-induced increase in microglial C/EBPbeta is prevented by coadministration of the MAP kinase inhibitors SB203580 (p38 inhibitor) + SP600125 (JNK inhibitor) or SB203580 + U0126 (ERK inhibitor). Systemic injection of LPS also increases brain nuclear levels of C/EBPbeta as shown by Western blot, and this increase is localized in microglial cells as shown by double immunofluorescence, in the first report to our knowledge of C/EBPbeta expression in activated glial cells in vivo. These findings support a role for C/EBPbeta in the activation of astrocytes and, particularly, microglia. Given the nature of the C/EBPbeta-regulated genes, we hypothesize that this factor participates in neurotoxic effects associated with glial activation. (c) 2006 Wiley-Liss, Inc.