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A novel labeling approach identifies three stability levels of acetylcholine receptors in the mouse neuromuscular junction in vivo.


ABSTRACT:

Background

The turnover of acetylcholine receptors at the neuromuscular junction is regulated in an activity-dependent manner. Upon denervation and under various other pathological conditions, receptor half-life is decreased.

Methodology/principal findings

We demonstrate a novel approach to follow the kinetics of acetylcholine receptor lifetimes upon pulse labeling of mouse muscles with ¹²?I-?-bungarotoxin in vivo. In contrast to previous assays where residual activity was measured ex vivo, in our setup the same animals are used throughout the whole measurement period, thereby permitting a dramatic reduction of animal numbers at increased data quality. We identified three stability levels of acetylcholine receptors depending on the presence or absence of innervation: one pool of receptors with a long half-life of ?13 days, a second with an intermediate half-life of ?8 days, and a third with a short half-life of ?1 day. Data were highly reproducible from animal to animal and followed simple exponential terms. The principal outcomes of these measurements were reproduced by an optical pulse-labeling assay introduced recently.

Conclusions/significance

A novel assay to determine kinetics of acetylcholine receptor turnover with small animal numbers is presented. Our data show that nerve activity acts on muscle acetylcholine receptor stability by at least two different means, one shifting receptor lifetime from short to intermediate and another, which further increases receptor stability to a long lifetime. We hypothesize on possible molecular mechanisms.

SUBMITTER: Strack S 

PROVIDER: S-EPMC3107218 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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A novel labeling approach identifies three stability levels of acetylcholine receptors in the mouse neuromuscular junction in vivo.

Strack Siegfried S   Petersen Yvonne Y   Wagner Anika A   Röder Ira V IV   Albrizio Marina M   Reischl Markus M   Wacker Irene U IU   Wilhelm Christoph C   Rudolf Rüdiger R  

PloS one 20110602 6


<h4>Background</h4>The turnover of acetylcholine receptors at the neuromuscular junction is regulated in an activity-dependent manner. Upon denervation and under various other pathological conditions, receptor half-life is decreased.<h4>Methodology/principal findings</h4>We demonstrate a novel approach to follow the kinetics of acetylcholine receptor lifetimes upon pulse labeling of mouse muscles with ¹²⁵I-α-bungarotoxin in vivo. In contrast to previous assays where residual activity was measure  ...[more]

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