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The Journal of Neuroscience, June 15, 2003, 23(12):5161-5169

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Aberrant Development of Motor Axons and Neuromuscular Synapses in MyoD-Null Mice

Zuo-Zhong Wang,^1 * Charles H. Washabaugh,^2 * Yun Yao,¹ Jun-Mei Wang,¹ Lili Zhang,¹ Martin P. Ontell,² Simon C. Watkins,² Michael A. Rudnicki,³ and Marcia Ontell²

Departments of ¹Neurobiology and ²Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, and ³Department of Molecular Medicine, Ottawa Health Research Unit, Ottawa Hospital, Ontario, Canada K1Y4E9

Myogenic regulatory factors (MRFs), muscle-specific transcription factors, are implicated in the activity-dependent regulation of nicotinic acetylcholine receptor (AChR) subunit genes. Here we show, with immunohistochemistry, Western blotting, and electron microscopy that MyoD, a member of the MRF family, also plays a role in fetal synapse formation. In the diaphragm of 14.5 d gestation (E14.5) wild-type and MyoD^-/- mice, AChR clusters (the formation of which is under a muscle intrinsic program) are confined to a centrally located endplate zone. This distribution persists in wild-type adult muscles. However, beginning at E15.5 and extending to the adult, innervated AChR clusters are distributed all over the diaphragm of MyoD^-/- mice, extending as far as the insertion of the diaphragm into the ribs. In wild-type muscle, motor axons terminate on clusters adjacent to the main intramuscular nerve; in MyoD^-/- muscle, axonal bundles form extensive secondary branches that terminate on the widely distributed clusters. The number of AChR clusters on adult MyoD^-/- and wild-type diaphram muscles is similar. Junctional fold density is reduced at MyoD^-/- endplates, and the transition from the fetal (, , , ) to adult-type (, , , ) AChRs is markedly delayed. However, MyoD^-/- mice assemble a complex postsynaptic apparatus that includes muscle-specific kinase (MuSK), rapsyn, erbB, and utrophin.

Key words: MyoD; knock-out mice; muscle development; neuromuscular junction; nicotinic acetylcholine receptors; MuSK

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Received Oct. 21, 2002; revised Apr. 1, 2003; accepted Apr. 3, 2003.

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