Restoration of synapse formation in<i>Musk</i>mutant mice expressing a Musk/Trk chimeric receptor

Herbst, Ruth; Avetisova, Ekaterina; Burden, Steven J.

doi:10.1242/dev.00112

Cited by 60 publications

(44 citation statements)

References 49 publications

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“…Rat MuSK-GFP cDNA was excised from L29/MuSK-FLAG-GFP (Herbst et al, 2002) with EcoRI and subcloned into pSG5, containing the HSA promoter cassette. The HSA::MuSK sequence, as well as the linked rabbit β-globin intron and SV40 polyadenylation signal, were released from the pSG5 plasmid, using SfiI and XmnI, and the gel-purified fragment was injected into zygotic male pronuclei, as described previously (Simon et al, 1992).…”

Section: Experimental Procedures Generation Of Hsa::musk Transgenic Micementioning

confidence: 99%

“…HSA::MuSK (MuSK-H or MuSK-L) mice were genotyped by PCR, as described previously for MCK::MuSK mice (Herbst et al, 2002). HB9 cre and Isl2 DTA mice have been described previously (Arber et al, 1999;Yang et al, 2001) and were genotyped by PCR (HB9 cre : 5′-CCGGTGAACGTGCAAAACAGGCTCTA-3′ and 5′-CTTCCAGGGCGCGAGTTGATAGC-3′; Isl2 DTA : 5′-ACGACGCTGCGGGATACTCT-3′ and 5′-CAACGCTAGAACTCCCCTCA-3′).…”

Section: Mouse Strains and Genotypingmentioning

confidence: 99%

“…HB9 cre and Isl2 DTA mice have been described previously (Arber et al, 1999;Yang et al, 2001) and were genotyped by PCR (HB9 cre : 5′-CCGGTGAACGTGCAAAACAGGCTCTA-3′ and 5′-CTTCCAGGGCGCGAGTTGATAGC-3′; Isl2 DTA : 5′-ACGACGCTGCGGGATACTCT-3′ and 5′-CAACGCTAGAACTCCCCTCA-3′). MuSK mutant mice were genotyped by PCR, as described previously (DeChiara et al, 1996;Herbst et al, 2002). Agrin ΔZ mice, lacking neural Agrin (Gautam et al, 1996), were genotyped by PCR (5′-GTCAGTGGGGGACCTAGAAAC-3′ and 5′-GTTGCTCTGCAGCGCCTT-3′).…”

Section: Mouse Strains and Genotypingmentioning

confidence: 99%

“…Intercostal muscles from wild-type, MuSK-L and MuSK-H mice were fixed (4% formaldehyde in PBS) overnight at 4°C, dehydrated in methanol, digested for 30 min with 20 μg/ml proteinase K, hybridized with digoxigenin-labeled riboprobes directed against mRNAs encoding the AChR δ subunit (Simon et al, 1992) or MuSK (Herbst et al, 2002) and processed as described previously (Yang et al, 2001). Weak, uniform staining was observed with sense probe for AChR δ subunit and MuSK (data not shown).…”

Section: Whole Mount In Situ-hybridizationmentioning

confidence: 99%

“…AChR cluster size and density was measured, as described previously (Herbst et al, 2002;Jaworski and Burden, 2006). Briefly, we stained P0 diaphragm muscle from wild-type, MuSK-H and MuSK-L mice with Alexa-594-α-BGT, and collected confocal image stacks at the same subsaturating amplifier gain for all genotypes.…”

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MuSK controls where motor axons grow and form synapses

2007

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SummaryIt had long been thought that motor axons approach muscles that are regionally unspecialized and induce postsynaptic differentiation by releasing signals that focally initiate transcriptional and post-translational responses in muscle. This neuro-centric view of synapse formation, however, has been challenged by recent experiments, which showed that AChR clusters are concentrated in the central region of muscle independent of innervation. This nerve-independent prepattern of AChR expression requires MuSK, a receptor tyrosine kinase that is critical for synapse formation. How muscle prepatterning is established and whether motor axons recognize this prepattern are not known. Here, we show that MuSK itself is prepatterned in muscle and that high, ectopic MuSK expression is sufficient to promote ectopic motor axon growth and synapse formation, indicating that the muscle prepattern is recognized by motor axons and promotes synapse formation in the central region of developing mammalian muscle. Further, we provide evidence that early expression of MuSK in developing myotubes is sufficient to re-establish muscle prepatterning independent of the MuSK promoter. Moreover, we show that ectopic MuSK expression stimulates synapse formation in the absence of Agrin and rescues the neonatal lethality of agrin mutant mice, demonstrating that MuSK, independent of Agrin, is sufficient to direct presynaptic and postsynaptic differentiation. In contrast to a neuro-centric view for synapse formation, these data demonstrate that the postsynaptic cell can play a dominant role in regulating synapse formation.

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Section: Experimental Procedures Generation Of Hsa::musk Transgenic Micementioning

confidence: 99%

Section: Mouse Strains and Genotypingmentioning

confidence: 99%

Section: Mouse Strains and Genotypingmentioning

confidence: 99%

Section: Whole Mount In Situ-hybridizationmentioning

confidence: 99%

mentioning

confidence: 99%

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MuSK controls where motor axons grow and form synapses

2007

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Phosphoinositide 3‐kinase acts through RAC and Cdc42 during agrin‐induced acetylcholine receptor clustering

Nizhynska¹,

Neumueller²,

Herbst³

2007

Developmental Neurobiology

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The formation of the neuromuscular junction (NMJ) is regulated by the nerve-derived heparan sulfate proteoglycan agrin and the muscle-specific kinase MuSK. Agrin induces a signal transduction pathway via MuSK, which promotes the reorganization of the postsynaptic muscle membrane. Activation of MuSK leads to the phosphorylation and redistribution of acetylcholine receptors (AChRs) and other postsynaptic proteins to synaptic sites. The accumulation of high densities of AChRs at postsynaptic regions represents a hallmark of NMJ formation and is required for proper NMJ function. Here we show that phosphoinositide 3-kinase (PI3-K) represents a component of the agrin/MuSK signaling pathway. Muscle cells treated with specific PI3-K inhibitors are unable to form full-size AChR clusters in response to agrin and AChR phosphorylation is reduced. Moreover, agrin-induced activation of Rac and Cdc42 is impaired in the presence of PI3-K inhibitors. PI3-K is localized to the postsynaptic muscle membrane consistent with a role during agrin/MuSK signaling. These results put PI3-K downstream of MuSK as regulator of AChR phosphorylation and clustering. Its role during agrin-stimulated Rac and Cdc42 activation suggests a critical function during cytoskeletal reorganizations, which lead to the redistribution of actin-anchored AChRs.

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Role of extracellular matrix proteins and their receptors in the development of the vertebrate neuromuscular junction

Singhal

Martin

2011

Developmental Neurobiology

122

107

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The vertebrate neuromuscular junction remains the best-studied model for understanding the mechanisms involved in synaptogenesis, due to its relatively large size, its simplicity of patterning and its unparalleled experimental accessibility. During neuromuscular development, each skeletal myofiber secretes and deposits around its extracellular surface an assemblage of extracellular matrix (ECM) proteins that ultimately form a basal lamina. This is also the case at the neuromuscular junction, where the motor nerve contributes additional factors. Before most of the current molecular components were known, it was clear that the synaptic ECM of adult skeletal muscles was unique in composition and contained factors sufficient to induce the differentiation of both pre- and postsynaptic membranes. Biochemical, genetic and microscopy studies have confirmed that agrin, laminin (221, 421, and 521), collagen IV (α3-α6), collagen XIII, perlecan and the ColQ-bound form of acetylcholinesterase are all synaptic ECM proteins with important roles in neuromuscular development. The roles of their many potential receptors and/or binding proteins has been more difficult to assess at the genetic level due to the complexity of membrane interactions with these large proteins, but roles for MuSK-LRP4 in agrin signaling and for integrins, dystroglycan, and voltage-gated calcium channels in laminin-dependent phenotypes have been identified. Synaptic extracellular matrix proteins and their receptors are involved in almost all aspects of synaptic development, including synaptic initiation, topography, ultrastructure, maturation, stability and transmission.

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Restoration of synapse formation inMuskmutant mice expressing a Musk/Trk chimeric receptor

Cited by 60 publications

References 49 publications

MuSK controls where motor axons grow and form synapses

MuSK controls where motor axons grow and form synapses

Phosphoinositide 3‐kinase acts through RAC and Cdc42 during agrin‐induced acetylcholine receptor clustering

Role of extracellular matrix proteins and their receptors in the development of the vertebrate neuromuscular junction

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