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Caffeine and nicotine decrease acetylcholine receptor clustering in C2C12 myotube culture

Abstract

As motor neurons approach skeletal muscle during development, agrin is released and induces acetylcholine receptor (AChR) clustering. Our laboratory investigates the effect of environmental agents on skeletal muscle development by using C2C12 cell culture. For the current project, we investigated both short-term and long-term exposure to caffeine, nicotine, or both, at physiologically relevant concentrations. Short-term exposure was limited to the last 48 h of myotube formation, whereas a long-term exposure of 2 weeks allowed for several generations of myoblast proliferation followed by myotube formation. Both agrin-induced and spontaneous AChR clustering frequencies were assessed. For agrin-induced AChR clustering, agrin was added for the last 16 h of myotube formation. Caffeine, nicotine, or both significantly decreased agrin-induced AChR clustering during short-term and long-term exposure. Furthermore, caffeine, nicotine, or both significantly decreased spontaneous AChR clustering during long-term, but not short-term exposure. Surprisingly, caffeine and nicotine in combination did not decrease AChR clustering beyond the effect of either treatment alone. We conclude that physiologically relevant concentrations of caffeine or nicotine decrease AChR clustering. Moreover, we predict that fetuses exposed to caffeine or nicotine may be less likely to form appropriate neuromuscular synapses.

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Acknowledgments

We thank Dr. James Stoehr and Dr. Carleton Jones for their insightful commentary in the creating and editing of this manuscript.

Author information

Correspondence to Wade A. Grow.

Additional information

This work was supported in part by the Office of Research and Sponsored Programs at Midwestern University, which provided intramural funding, and by the Physician Assistant Program, which provided additional funding.

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Kordosky-Herrera, K., Grow, W.A. Caffeine and nicotine decrease acetylcholine receptor clustering in C2C12 myotube culture. Cell Tissue Res 335, 341–348 (2009). https://doi.org/10.1007/s00441-008-0691-8

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Keywords

  • Acetylcholine receptor
  • Agrin
  • Caffeine
  • Nicotine
  • C2C12 cell culture (mouse)