AChR channel conversion and AChR-adjusted neuronal survival during embryonic development

Yampolsky, Pessah; Gensler, Sven; McArdle, Joseph J.; Witzemann, Veit

doi:10.1016/j.mcn.2007.12.014

Cited by 21 publications

(42 citation statements)

References 41 publications

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“…Using this method we observe that the loss of AChR from onetime labeled end plates results in a decrease of r-bgt-stained end plate area, supporting the observation that newly synthesized receptors are added to the periphery of end plates (15). Therefore, it is possible to follow changes in AChR stability by quantifying end plate areas over time instead of fluorescence intensities, which are more prone to experimental variation.…”

Section: Discussionsupporting

confidence: 68%

“…Second, we proved previously the validity of the area analysis approach for quantification of AChR dynamics, by showing that newly synthesized AChR incorporated into the end plate surface in a unidirectional fashion, proceeding from the periphery to the center (15,29 For staining, sections were moistened with PBS and permeabilized with 0.1% Triton X-100 for 10 min. After washing with PBS, slices were blocked with 10% FBS/PBS for 20 min.…”

Section: Surgical Procedures-newborn Achrmentioning

confidence: 63%

“…with Wild Type and AChR⑀ Ϫ/Ϫ Lines-The estimated mean total content of adult AChR⑀ (Ͻ50%) in wild type animals on P6 is significantly lower than the AChR⑀ content (75.8 Ϯ 3.8%) in end plates of AChR ␥-GFP/␥-GFP animals on the same postnatal day (15,16). To address the possible influence of the untimely ␥-to-⑀ conversion in the AChR ␥-GFP/␥-GFP line on end plate receptor stabilization we compared animals 6 days of age from both wild type and AChR ␥-GFP/␥-GFP lines.…”

Section: ␥-Gfp/␥-gfpmentioning

confidence: 95%

“…was generated by homologous recombination fusing GFP into the ␥ subunit gene as described previously (15,31). GFP-labeled ␥ subunits, ␥ -GFP subunits, are assembled into AChR␥ -GFP complexes that substitute the wild type ␥ subunit-containing AChR␥.…”

Section: ␥-Gfp/␥-gfpmentioning

confidence: 99%

“…This is based on our observation that new receptors are added to the end plates from the periphery (15). So, after the initial labeling, new nonfluorescent AChRs are inserted, and the area of fluorescent receptors will decrease as a consequence of AChR loss.…”

Section: Time Lapse In Vivo Imaging At Early Postnatal Stages-ourmentioning

confidence: 99%

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Time Lapse in Vivo Visualization of Developmental Stabilization of Synaptic Receptors at Neuromuscular Junctions

Yampolsky

Pacifici

Lomb

et al. 2010

Journal of Biological Chemistry

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The lifetime of nicotinic acetylcholine receptors (AChRs) in neuromuscular junctions (NMJs) is increased from <1 day to >1 week during early postnatal development. However, the exact timing of AChR stabilization is not known, and its correlation to the concurrent embryonic to adult AChR channel conversion, NMJ remodeling, and neuromuscular diseases is unclear. Using a novel time lapse in vivo imaging technology we show that replacement of the entire receptor population of an individual NMJ occurs end plate-specifically within hours. This makes it possible to follow directly in live animals changing stabilities of end plate receptors. In three different, genetically modified mouse models we demonstrate that the metabolic half-life values of synaptic AChRs increase from a few hours to several days after postnatal day 6. Developmental stabilization is independent of receptor subtype and apparently regulated by an intrinsic muscle-specific maturation program. Myosin Va, an F-actin-dependent motor protein, is also accumulated synaptically during postnatal development and thus could mediate the stabilization of end plate AChR.

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Section: Discussionsupporting

confidence: 68%

Section: Surgical Procedures-newborn Achrmentioning

confidence: 63%

Section: ␥-Gfp/␥-gfpmentioning

confidence: 95%

Section: ␥-Gfp/␥-gfpmentioning

confidence: 99%

Section: Time Lapse In Vivo Imaging At Early Postnatal Stages-ourmentioning

confidence: 99%

See 3 more Smart Citations

Time Lapse in Vivo Visualization of Developmental Stabilization of Synaptic Receptors at Neuromuscular Junctions

Yampolsky

Pacifici

Lomb

et al. 2010

Journal of Biological Chemistry

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Postsynaptic Development of the Neuromuscular Junction in Mice Lacking the γ-Subunit of Muscle Nicotinic Acetylcholine Receptor

et al. 2009

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The mammalian muscle nicotinic acetylcholine receptor (AChR) is composed of five membranespanning subunits and its composition differs between embryonic and adult muscles. In embryonic muscles, it is composed of two α-, one β-, one δ-, and one γ-subunit; the γ-subunit is later replaced by the ε-subunit during postnatal development. This unique temporal expression pattern of the γ-subunit suggests it may play specific roles in embryonic muscles. To address this issue, we examined the formation and function of the neuromuscular junction in mouse embryos deficient in the γ-subunit. At embryonic day 15.5, AChR clusters were absent and the spontaneous miniature endplate potentials were undetectable in the mutant muscles. However, electrical stimulation of the nerves triggered muscle contraction and elicited postsynaptic endplate potential (EPP) in the mutant muscles, although the magnitude of the muscle contraction and the amplitudes of the EPPs were smaller in the mutant compared to the wild-type muscles. Reintroducing a wild-type γ-subunit into the mutant myotubes restored the formation of AChR clusters in vitro. Together, these results have demonstrated that functional AChRs were present in the mutant muscle membrane, but at reduced levels. Thus, in the absence of the γ-subunit, a combination of α, β, and δ subunits may assemble into functional receptors in vivo. These results also suggest that the γ-subunit maybe involved in interacting with rapsyn, a cytoplasmic protein required for AChR clustering.

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Cloning and characterization of nicotinic acetylcholine receptor γ-like gene in adult transparent Pristella maxillaris

Chen

Zhou

et al. 2021

Gene

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AChR channel conversion and AChR-adjusted neuronal survival during embryonic development

Cited by 21 publications

References 41 publications

Time Lapse in Vivo Visualization of Developmental Stabilization of Synaptic Receptors at Neuromuscular Junctions

Time Lapse in Vivo Visualization of Developmental Stabilization of Synaptic Receptors at Neuromuscular Junctions

Postsynaptic Development of the Neuromuscular Junction in Mice Lacking the γ-Subunit of Muscle Nicotinic Acetylcholine Receptor

Cloning and characterization of nicotinic acetylcholine receptor γ-like gene in adult transparent Pristella maxillaris

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