Neuromuscular synaptic function in mice lacking major subsets of gangliosides

Zitman, Femke M.P.; Todorov, Boyan; Jacobs, Bart C.; Verschuuren, Jan J.; Furukawa, Koichi; Willison, Hugh J.; Plomp, Jaap J.

doi:10.1016/j.neuroscience.2008.08.034

Cited by 24 publications

(29 citation statements)

References 65 publications

(86 reference statements)

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“…1), most likely at highly accumulated level (Inoue et al, 2002). We here acutely removed the sialic acid component of this only remaining ganglioside at dKO NMJs, thereby creating a ganglioside-free nerve terminal, and studied the effect on transmitter release.Male and female dKO mice (Inoue et al, 2002;Zitman et al, 2008) were used in the experiments, at ~11 weeks of age and 22.9 ± 1.1 g body weight. Mice were killed by CO 2 asphyxiation.…”

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confidence: 99%

“…Male and female dKO mice (Inoue et al, 2002;Zitman et al, 2008) were used in the experiments, at ~11 weeks of age and 22.9 ± 1.1 g body weight. Mice were killed by CO 2 asphyxiation.…”

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confidence: 99%

“…1), we have previously shown that presence of the complete family of gangliosides is not strictly required for functional synaptic transmission. Electrophysiological measurement of acetylcholine (ACh) release at NMJs of GM2/GD2-synthase knockout (GM2s-KO), GD3-synthase knockout (GD3s-KO), or compound null-mutant mice lacking both GM2/GD2-and GD3-synthase (dKO), showed only subtle changes and no failure of synaptic transmission was observed (Bullens et al, 2002;Zitman et al, 2008;Zitman et al, 2009). This suggests a fine-tuning role for gangliosides in transmitter release, rather than their presence being an absolute requirement.…”

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confidence: 99%

“…Subsequently, μ-conotoxin-GIIIB (3 μM; Scientific Marketing Associates, Barnet, Herts, UK) was added to block voltage-gated Na + channels on the muscle membrane in order to prevent muscle action potentials and contraction. ACh release at NMJs at room temperature (20-22 C) was assessed using an intracellular electrophysiological method, as described before (Zitman et al, 2008) 2C-E). For comparison, EPP rundown level at wild-type NMJs under comparable experimental conditions in our previous study was 80.9  1.2% (Zitman et al, 2008).…”

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confidence: 99%

“…ACh release at NMJs at room temperature (20-22 C) was assessed using an intracellular electrophysiological method, as described before (Zitman et al, 2008) 2C-E). For comparison, EPP rundown level at wild-type NMJs under comparable experimental conditions in our previous study was 80.9  1.2% (Zitman et al, 2008).…”

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Total ganglioside ablation at mouse motor nerve terminals alters neurotransmitter release level

Zitman

Todorov

Furukawa

et al. 2009

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However, at the neuromuscular junction (NMJ) of transgenic mice lacking major subsets of gangliosides ( Fig. 1), we have previously shown that presence of the complete family of gangliosides is not strictly required for functional synaptic transmission. Electrophysiological measurement of acetylcholine (ACh) release at NMJs of GM2/GD2-synthase knockout (GM2s-KO), GD3-synthase knockout (GD3s-KO), or compound null-mutant mice lacking both GM2/GD2-and GD3-synthase (dKO), showed only subtle changes and no failure of synaptic transmission was observed (Bullens et al., 2002;Zitman et al., 2008;Zitman et al., 2009). This suggests a fine-tuning role for gangliosides in transmitter release, rather than their presence being an absolute requirement. Alternatively, the different types of gangliosides might act mutually compensatory in supporting transmitter release. Such a compensatory action of GM3 ganglioside might explain the redundancy we found for the other gangliosides, because in all three investigated transgenic strains at least this ganglioside remains present in the neural membrane (Fig. 1), most likely at highly accumulated level (Inoue et al., 2002). We here acutely removed the sialic acid component of this only remaining ganglioside at dKO NMJs, thereby creating a ganglioside-free nerve terminal, and studied the effect on transmitter release.Male and female dKO mice (Inoue et al., 2002;Zitman et al., 2008) were used in the experiments, at ~11 weeks of age and 22.9 ± 1.1 g body weight. Mice were killed by CO 2 asphyxiation. Left and right hemidiaphragms with phrenic nerves were dissected and pinned out in Ringer's medium (119 mM NaCl, 4.5 mM KCl, 2 mM CaCl 2 , 1 mM MgSO 4 , 1mM Zitman et al. Gangliosides and neurotransmitter release3 NaH 2 PO 4 , 23 mM NaHCO 3 , 11 mM glucose, pH 7.4), pre-gassed with 95% O 2 /5% CO 2 .Animal experiments were carried out according to Dutch law and Leiden University guidelines.dKO phrenic nerve-hemidiaphragm preparations (6 left-and 4 right-hemidiaphragms from n=10 dKO mice) were treated with 2 units neuraminidase type V from Clostridium perfringens (Sigma-Aldrich, Zwijndrecht, The Netherlands) in 1 ml Ringer's medium for 1 h at 32 °C. This disrupts GM3 by cleaving the α2,3-link between the sialic acid and galactose molecule (Perillo et al., 1994). We used tissue from dKO mice because neuraminidase treatment of wild-type tissue will not induce complete ganglioside ablation due to the inability of the neuraminidase to desialylate GM1 (Perillo et al., 1994). The contralateral hemidiaphragm from each mouse was incubated as a control in Ringer's medium only.Preparations were then rinsed in Ringer's medium for 10 min. Subsequently, μ-conotoxin- These results show that acute absence of all gangliosides by in vitro disruption of GM3, the only remaining type of ganglioside at dKO NMJs, at the one hand increases presynaptic neurotransmitter release by enhancing the amount of ACh released per nerve impulse but, on the other hand, reduces release by causing a more pronounced rundown during ...

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“…Male and female dKO mice (Inoue et al, 2002;Zitman et al, 2008) were used in the experiments, at ~11 weeks of age and 22.9 ± 1.1 g body weight. Mice were killed by CO 2 asphyxiation.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Total ganglioside ablation at mouse motor nerve terminals alters neurotransmitter release level

Zitman

Todorov

Furukawa

et al. 2009

Synapse

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Quantal Analysis of Endplate Potentials in Mouse Flexor Digitorum Brevis Muscle

Ribchester

2011

CP Mouse Biology

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The isolated flexor digitorum brevis (FDB) muscle from mice is extremely well suited to rapid acquisition of data and analysis of neurotransmitter release and action at neuromuscular junctions, because the muscle and its tibial nerve supply are simple to dissect and its constituent muscle fibers are short (<1 mm) and isopotential along their length. Methods are described here for dissection of FDB, stimulation of the tibial nerve, microelectrode recording from individual muscle fibers, and quantal analysis of endplate potentials (EPPs) and miniature endplate potentials (MEPPs). Curr. Protoc. Mouse Biol. 1:429-444 © 2011 by John Wiley & Sons, Inc.

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Sphingolipids regulate neuromuscular synapse structure and function in Drosophila

West

Briggs

Fjeldstad

et al. 2018

J of Comparative Neurology

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Sphingolipids are found in abundance at synapses and have been implicated in regulation of synapse structure, function, and degeneration. Their precise role in these processes, however, remains obscure. Serine Palmitoyl‐transferase (SPT) is the first enzymatic step for synthesis of sphingolipids. Analysis of the Drosophila larval neuromuscular junction (NMJ) revealed mutations in the SPT enzyme subunit, lace/SPTLC2 resulted in deficits in synaptic structure and function. Although NMJ length is normal in lace mutants, the number of boutons per NMJ is reduced to ∼50% of the wild type number. Synaptic boutons in lace mutants are much larger but show little perturbation to the general ultrastructure. Electrophysiological analysis of lace mutant synapses revealed strong synaptic transmission coupled with predominance of depression over facilitation. The structural and functional phenotypes of lace mirrored aspects of Basigin (Bsg), a small Ig‐domain adhesion molecule also known to regulate synaptic structure and function. Mutant combinations of lace and Bsg generated large synaptic boutons, while lace mutants showed abnormal accumulation of Bsg at synapses, suggesting that Bsg requires sphingolipid to regulate structure of the synapse. In support of this, we found Bsg to be enriched in lipid rafts. Our data points to a role for sphingolipids in the regulation and fine‐tuning of synaptic structure and function while sphingolipid regulation of synaptic structure may be mediated via the activity of Bsg.

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Neuromuscular synaptic function in mice lacking major subsets of gangliosides

Cited by 24 publications

References 65 publications

Total ganglioside ablation at mouse motor nerve terminals alters neurotransmitter release level

Total ganglioside ablation at mouse motor nerve terminals alters neurotransmitter release level

Quantal Analysis of Endplate Potentials in Mouse Flexor Digitorum Brevis Muscle

Sphingolipids regulate neuromuscular synapse structure and function in Drosophila

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