Neurotransmitter Acetylcholine Negatively Regulates Neuromuscular Synapse Formation by a Cdk5-Dependent Mechanism
ABSTRACT Synapse formation requires interactions between pre- and postsynaptic cells to establish the connection of a presynaptic nerve terminal with the neurotransmitter receptor-rich postsynaptic apparatus. At developing vertebrate neuromuscular junctions, acetylcholine receptor (AChR) clusters of nascent postsynaptic apparatus are not apposed by presynaptic nerve terminals. Two opposing activities subsequently promote the formation of synapses: positive signals stabilize the innervated AChR clusters, whereas negative signals disperse those that are not innervated. Although the nerve-derived protein agrin has been suggested to be a positive signal, the negative signals remain elusive. Here, we show that cyclin-dependent kinase 5 (Cdk5) is activated by ACh agonists and is required for the ACh agonist-induced dispersion of the AChR clusters that have not been stabilized by agrin. Genetic elimination of Cdk5 or blocking ACh production prevents the dispersion of AChR clusters in agrin mutants. Therefore, we propose that ACh negatively regulates neuromuscular synapse formation through a Cdk5-dependent mechanism.
- SourceAvailable from: Giancarlo V. De Ferrari
[Show abstract] [Hide abstract]
- "In this regard, neural-and muscle-derived molecules have been described to play such inhibitory roles at the vertebrate NMJ. One the one hand, the neurotransmitter ACh displays an AChRdisaggregating activity at the most abundant non-innervated ( " extrasynaptic " ) domains of the muscle membrane at nascent NMJs (Lin et al., 2005; Misgeld et al., 2005). ACh acts through the cyclin-dependent kinase 5 (Cdk5) which phosphorylates the intermediate filament protein nestin, that becomes dissociated from the cytoskeletal network and is subsequently degraded (Fu et al., 2005; Yang et al., 2011). "
ABSTRACT: Cumulative evidence indicates that Wnt pathways play crucial and diverse roles to assemble the neuromuscular junction (NMJ), a peripheral synapse characterized by the clustering of acetylcholine receptors (AChR) on postsynaptic densities. The molecular determinants of Wnt effects at the NMJ are still to be fully elucidated. We report here that the Wnt receptor Frizzled-9 (Fzd9) is expressed in developing skeletal muscles during NMJ synaptogenesis. In cultured myotubes, gain-and loss-of-function experiments revealed that Fzd9-mediated signaling impairs the AChR-clustering activity of agrin, an organizer of postsynaptic differentiation. Overexpression of Fzd9 induced the cytosolic accumulation of β-catenin, a key regulator of Wnt signaling. Consistently, Fzd9 and β-catenin localize in the postsynaptic domain of embryonic NMJs in vivo. Our findings represent the first evidence pointing to a crucial role of a Fzd-mediated, β-catenin-dependent signaling on the assembly of the vertebrate NMJ.Frontiers in Cellular Neuroscience 04/2014; 8. DOI:10.3389/fncel.2014.00110 · 4.18 Impact Factor
[Show abstract] [Hide abstract]
- "There is a careful balance between signals promoting and inhibiting postsynaptic stabilisation. A compromised presynaptic terminal may no longer provide enough positive signals, such as AGRN (McMahan, 1990; Misgeld et al. 2005), that stabilise the clustering of AChR at the postsynaptic side, to counteract a prolonged exposure to ACh, which is also a signal for AChR cluster dissipation (Lin et al. 2005; Misgeld et al. 2005). "
ABSTRACT: MuSK myasthenia gravis is a rare, severe autoimmune disease of the neuromuscular junction, only identified in 2001, with unclear pathogenic mechanisms. In this review we describe the clinical aspects that distinguish MuSK MG from AChR MG, review what is known about the role of MuSK in the development and function of the neuromuscular junction, and discuss the data that address how the antibodies to MuSK lead to neuromuscular transmission failure.Journal of Anatomy 03/2013; 224(1). DOI:10.1111/joa.12034 · 2.23 Impact Factor
[Show abstract] [Hide abstract]
- "These results suggest that the active zone density at NMJs is independent of the synaptic activity. This contrasts with the role of synaptic activity in synapse elimination and the postsynaptic differentiation of NMJs (Lichtman et al., 1985; Buffelli et al., 2003; Lin et al., 2005; Misgeld et al., 2005). Analysis using the three-dimensional reconstruction of transmission electron micrographs shows that the active zone density of Drosophila NMJs is also maintained in a homeostatic manner even after manipulations to augment or attenuate synaptic efficiency (Meinertzhagen et al., 1998; Reiff et al., 2002). "
ABSTRACT: Neural circuits transmit information through synapses, and the efficiency of synaptic transmission is closely related to the density of presynaptic active zones, where synaptic vesicles are released. The goal of this review is to highlight recent insights into the molecular mechanisms that control the number of active zones per presynaptic terminal (active zone density) during developmental and stimulus-dependent changes in synaptic efficacy. At the neuromuscular junctions (NMJs), the active zone density is preserved across species, remains constant during development, and is the same between synapses with different activities. However, the NMJ active zones are not always stable, as exemplified by the change in active zone density during acute experimental manipulation or as a result of aging. Therefore, a mechanism must exist to maintain its density. In the central nervous system (CNS), active zones have restricted maximal size, exist in multiple numbers in larger presynaptic terminals, and maintain a constant density during development. These findings suggest that active zone density in the CNS is also controlled. However, in contrast to the NMJ, active zone density in the CNS can also be increased, as observed in hippocampal synapses in response to synaptic plasticity. Although the numbers of known active zone proteins and protein interactions have increased, less is known about the mechanism that controls the number or spacing of active zones. The following molecules are known to control active zone density and will be discussed herein: extracellular matrix laminins and voltage-dependent calcium channels, amyloid precursor proteins, the small GTPase Rab3, an endocytosis mechanism including synaptojanin, cytoskeleton protein spectrins and β-adducin, and a presynaptic web including spectrins. The molecular mechanisms that organize the active zone density are just beginning to be elucidated.Frontiers in Molecular Neuroscience 02/2012; 5:12. DOI:10.3389/fnmol.2012.00012 · 4.08 Impact Factor