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The Journal of Neuroscience, May 30, 2007, 27(22):5948-5957; doi:10.1523/JNEUROSCI.0621-07.2007

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Development/Plasticity/Repair
Regulation of the Intermediate Filament Protein Nestin at Rodent Neuromuscular Junctions by Innervation and Activity

Hyuno Kang,¹ Le Tian,¹ Young-Jin Son,¹ Yi Zuo,¹ Diane Procaccino,¹ Flora Love,¹ Christopher Hayworth,¹ Joshua Trachtenberg,¹ Michelle Mikesh,¹ Lee Sutton,¹ Olga Ponomareva,¹ John Mignone,² Grigori Enikolopov,² Mendell Rimer,¹ and Wesley Thompson¹

¹Section of Neurobiology, Institute of Neuroscience, and Institute for Cell and Molecular Biology, University of Texas, Austin, Texas 78712, and ²Cold Spring Harbor Laboratories, Cold Spring Harbor, New York 11724

Correspondence should be addressed to Wesley Thompson, Section of Neurobiology, University of Texas, Austin, TX 78712. Email: wes{at}mail.utexas.edu

The intermediate filament nestin is localized postsynaptically at rodent neuromuscular junctions. The protein forms a filamentous network beneath and between the synaptic gutters, surrounds myofiber nuclei, and is associated with Z-discs adjacent to the junction. In situ hybridization shows that nestin mRNA is synthesized selectively by synaptic myonuclei. Although weak immunoreactivity is present in myelinating Schwann cells that wrap the preterminal axon, nestin is not detected in the terminal Schwann cells (tSCs) that cover the nerve terminal branches. However, after denervation of muscle, nestin is upregulated in tSCs and in SCs within the nerve distal to the lesion site. In contrast, immunoreactivity is strongly downregulated in the muscle fiber. Transgenic mice in which the nestin neural enhancer drives expression of a green fluorescent protein (GFP) reporter show that the regulation in SCs is transcriptional. However, the postsynaptic expression occurs through enhancer elements distinct from those responsible for regulation in SCs. Application of botulinum toxin shows that the upregulation in tSCs and the loss of immunoreactivity in muscle fibers occurs with blockade of transmitter release. Extrinsic stimulation of denervated muscle maintains the postsynaptic expression of nestin but does not affect the upregulation in SCs. Thus, a nestin-containing cytoskeleton is promoted in the postsynaptic muscle fiber by nerve-evoked muscle activity but suppressed in tSCs by transmitter release. Nestin antibodies and GFP driven by nestin promoter elements serve as excellent markers for the reactive state of SCs. Vital imaging of GFP shows that SCs grow a dynamic set of processes after denervation.

Key words: transgenic; synaptic gene expression; terminal Schwann cell; denervation; paralysis; muscle stimulation

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Received Feb. 12, 2007; revised April 20, 2007; accepted April 25, 2007.

Correspondence should be addressed to Wesley Thompson, Section of Neurobiology, University of Texas, Austin, TX 78712. Email: wes{at}mail.utexas.edu

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				F. A. Court, T. H. Gillingwater, S. Melrose, D. L. Sherman, K. N. Greenshields, A. J. Morton, J. B. Harris, H. J. Willison, and R. R. Ribchester Identity, developmental restriction and reactivity of extralaminar cells capping mammalian neuromuscular junctions J. Cell Sci., December 1, 2008; 121(23): 3901 - 3911. [Abstract] [Full Text] [PDF]

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