Abstract
Environmental, congenital, and acquired immunological insults perturbing neuromuscular junction (NMJ) activity may induce a variety of debilitating neuromuscular pathologies. However, the molecular elements linking NMJ dysfunction to long-term myopathies are unknown. Here, we report dramatically elevated levels of mRNA encoding c-Fos and the "readthrough" (R) variant of acetylcholinesterase (AChE) in muscles of transgenic mice overexpressing synaptic (S) AChE in motoneurons and in control mice treated with the irreversible cholinesterase inhibitor diisopropylfluorophosphonate (DFP). Tongue muscles from DFP-treated and AChE-S transgenic mice displayed exaggerated neurite branching and disorganized, wasting fibers. Moreover, diaphragm muscles from both transgenic and DFP-treated mice exhibited NMJ proliferation. 2′-O-methyl-protected antisense oligonucleotides targeted to AChE mRNA suppressed feedback upregulation of AChE and ameliorated DFP-induced NMJ proliferation. Our findings demonstrate common transcriptional responses to cholinergic NMJ stress of diverse origin, and implicate deregulated AChE expression in excessive neurite outgrowth, uncontrolled synaptogenesis, and myopathology.
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Andres C., Beeri R., Friedman A., Lev-Lehman E., Henis S., Timberg R., et al. (1997). Acetylcholinesterase-transgenic mice display embryonic modulations in spinal cord choline acetyltransferase and neurexin Ibeta gene expression followed by late-onset neuromotor deterioration. Proc. Natl. Acad. Sci. USA 94, 8173–8178.
Andres C., Seidman S., Beeri R., Timberg R., and Soreq H. (1998) Transgenic acetylcholinesterase induces enlargement of murine neuromuscular junctions but leaves spinal cord synapses intact. Neurochem Int. 32, 449–456.
Ben Aziz Aloya R., Seidman S., Timberg R., Sternfeld M., Zakut H., and Soreq H. (1993). Expression of a human acetylcholinesterase promoter-reporter construct in developing neuromuscular junctions of Xenopus embryos. Proc. Natl. Acad. Sci. USA 90, 2471–2475.
Chollat Namy A., Delamanche I. S., and Bouchaud C. (1993) Variation in the expression of c-fos after intoxication by soman. Comparative study using in situ hybridization and immunohistochemistry. Brain Res. 603, 32–37.
Darboux I., Barthalay Y., Piovant M., and Hipeau Jacquotte R. (1996) The structure-function relationships in Drosophila neurotactin show that cholinesterasic domains may have adhesive properties. EMBO J. 15, 4835–4843.
de la Escalera S., Bockamp E. O., Moya F., Piovant M., and Jimenez F. (1990) Characterization and gene cloning of neurotactin, a Drosophila transmembrane protein related to cholinesterases. EMBO J. 9, 3593–3601.
Donger C., Krejci E., Serradell A. P., Eymard B., Bon S., Nicole S., et al. (1998). Mutation in the human acetylcholinesterase-associated collagen gene, COLQ, is responsible for congenital myasthenic syndrome with end-plate acetylcholinesterase deficiency (Type Ic). Am. J. Hum. Genet. 63, 967–975.
Engel A. G., Lambert E. H., and Santa T. (1973) Study of long-term anticholinesterase therapy. Effects on neuromuscular transmission and on motor endplate fine structure. Neurology 23, 1273–1281.
Engelhardt J. I., Siklos L., and Appel S. H. (1997) Immunization of guinea pigs with human choline acetyltransferase induces selective lower motoneuron destruction. J. Neuroimmunol. 78, 57–68.
Evoli A., Batocchi A. P., and Tonali P. (1996) A practical guide to the recognition and management of myasthenia gravis. Drugs 52, 662–670.
Friedman A., Kaufer D., Shemer J., Hendler I., Soreq H., and Tur Kaspa I. (1996). Pyridostigmine brain penetration under stress enhances neuronal excitability and induces early immediate transcriptional response. Nat. Med. 2, 1382–1385.
Gibbs R. B. and Martynowski C. (1997). Nerve growth factor induces Fos-like immunoreactivity within identified cholinergic neurons in the adult rat basal forebrain. Brain Res. 753, 141–151.
Grifman M., Galyam N., Seidman S., and Soreq H. (1998) Functional redundancy of acetylcholinesterase and neuroligin in mammalian neuritogenesis. Proc. Natl. Acad. Sci. USA 95, 13,935–13,940.
Grisaru D., Lev-Lehman E., Shapira M., Chaikin E., Lessing J. B., Eldor A., et al. (1999) Human osteogenesis involves differentiation-dependent increases in the morphogenically active 3′ alternative splicing variant of acetylcholinesterase. Mol. Cell Biol. 19, 788–795.
Haley R. W., Kurt T. L., and Hom J. (1997) Is there a Gulf War Syndrome? Searching for syndromes by factor analysis of symptoms. JAMA 277, 215–222.
Hudson C. S., Rash J. E., Tiedt T. N., and Albuquerque E. X. (1978) Neostigmine-induced alterations at the mammalian neuromuscular junction. II. Ultrastructure. J. Pharmacol. Exp. Ther. 205, 340–356.
Ichtchenko K., Hata Y., Nguyen T., Ullrich B., Missler M., Moomaw C., and Sudhof T. C. (1995). Neuroligin 1: a splice site-specific ligand for betaneurexins. Cell 81, 435–443.
Jasmin B. J., Lee R. K., and Rotundo R. L. (1993) Compartmentalization of acetylcholinesterase mRNA and enzyme at the vertebrate neuromuscular junction. Neuron 11, 467–477.
Kaufer D., Friedman A., and Soreq H. (1999) The vicious circle of stress and anticholinesterase responses. Neuroscientist 5, 173–183.
Kaufer D., Friedman A., Seidman S., and Soreq H. (1998) Acute stress facilitates long-lasting changes in cholinergic gene expression. Nature 393, 373–377.
Kawabuchi M., Osame M., Watanabe S., Igata A., and Kanaseki T. (1976) Myopathic changes at the end-plate region induced by neostigmine methylsulfate. Experientia 32, 632–635.
Koenigsberger C., Chiappa S., and Brimijoin S. (1997) Neurite differentiation is modulated in neuroblastoma cells engineered for altered acetylcholinesterase expression. J. Neurochem. 69, 1389–1397.
Laskowski M. B., Olson W. H., and Dettbarn W. D. (1975) Ultrastructural changes at the motor end-plant produced by an irreversible cholinesterase inhibitor. Exp. Neurol. 47, 290–306.
Layer P. G. and Willbold E. (1995) Novel functions of cholinesterases in development, physiology and disease. Prog. Histochem. Cytochem. 29, 1–99.
Lindstrom J. (1998) Mutations causing muscle weakness. Proc. Natl. Acad. Sci. USA 95, 9070,9071.
Livneh A., Sarova I., Michaeli D., Pras M., Wagner K., Zakut H., and Soreq H. (1988). Antibodies against acetylcholinesterase and low levels of cholinesterases in a patient with an atypical neuromuscular disorder. Clin. Immunol. Immunopathol. 48, 119–131.
Massoulie J., Pezzementi L., Bon S., Krejci E., and Vallette F. M. (1993) Molecular and cellular biology of cholinesterases. Prog. Neurobiol. 41, 31–91.
Sanes J. R. and Lichtman J. W. (1999) Development of the vertebrate neuromuscular junction. Ann. Rev. Neurosci. 22, 389–442.
Schonbeck S., Chrestel S., and Hohlfeld R. (1990) Myasthenia gravis: prototype of the antireceptor autoimmune diseases. Int. Rev. Neurobiol. 32, 175–200.
Seidman S., Sternfeld M., Ben Aziz Aloya R., Timberg R., Kaufer Nachum D., and Soreq H. (1995). Synaptic and epidermal accumulations of human acetylcholinesterase are encoded by alternative 3′-terminal exons. Mol. Cell Biol. 15, 2993–3002.
Seidman S., Eckstein F., Grifman M., and Soreq H. Antisense technologies have a future combating neurodegenerative disease. Antisense Res. Drug Dev. 9, 333–340.
Shapira M., Seidman S., Sternfeld M., Timberg R., Kaufer D., Patrick J., and Soreq H. (1994). Transgenic engineering of neuromuscular junctions in Xenopus laevis embryos transiently overexpressing key cholinergic proteins. Proc. Natl. Acad. Sci. USA 91, 9072–9076.
Sharma K. V. and Bigbee J. W. (1998) Acetylcholinesterase antibody treatment results in neurite detachment and reduced outgrowth from cultured neurons: further evidence for a cell adhesive role for neuronal acetylcholinesterase. J. Neurosci. Res. 53, 454–464.
Slotkin T. A., McCook E. C., and Seidler F. J. (1997) Cryptic brain cell injury caused by fetal nicotine exposure is associated with persistent elevations of c-fos protooncogene expression. Brain Res. 750, 180–188.
Sternfeld M., Ming G., Song H., Sela K., Timberg R., Poo M., and Soreq H. (1998). Acetylcholinesterase enhances neurite growth and synapse development through alternative contributions of its hydrolytic capacity, core protein, and variable C termini. J. Neurosci. 18, 1240–1249.
Sternfeld M., Patrick J. D., and Soreq H. (1998) Position effect variegations and brain-specific silencing in transgenic mice overexpressing human acetylcholinesterase variants. J. Physiol. (Paris) 92, 249–255.
Swash M. (1975) Motor innervation of myasthenic muscles (Letter). Lancet 2, 663.
Taylor P. (1996) Agents acting at the neuromuscular junction and autonomic ganglia, in Goodman and Gilman’s The Pharmacological Basis of Therapeutics (Hardman J. G., Limbird L. E., Molinoff P. B., and Ruddon R. W., eds.), McGraw-Hill, New York, pp. 177–197.
Taylor P., Radic Z., Hosea N. A., Camp S., Marchot P., and Berman H. A. (1995). Structural bases for the specificity of cholinesterase catalysis and inhibition. Toxicol. Lett. 82–83, 453–458.
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Lev-Lehman, E., Evron, T., Broide, R.S. et al. Synaptogenesis and myopathy under acetylcholinesterase overexpression. J Mol Neurosci 14, 93–105 (2000). https://doi.org/10.1385/JMN:14:1-2:093
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DOI: https://doi.org/10.1385/JMN:14:1-2:093