Optogenetic control of contractile function in skeletal muscle

Optogenetic stimulation allows activation of cells with high spatial and temporal precision. Here we show direct optogenetic stimulation of skeletal muscle from transgenic mice expressing the light-sensitive channel Channelrhodopsin-2 (ChR2). Largest tetanic contractions are observed with 5-ms light pulses at 30 Hz, resulting in 84% of the maximal force induced by electrical stimulation. We demonstrate the utility of this approach by selectively stimulating with a light guide individual intralaryngeal muscles in explanted larynges from ChR2-transgenic mice, which enables selective opening and closing of the vocal cords. Furthermore, systemic injection of adeno-associated virus into wild-type mice provides sufficient ChR2 expression for optogenetic opening of the vocal cords. Thus, direct optogenetic stimulation of skeletal muscle generates large force and provides the distinct advantage of localized and cell-type-specific activation. This technology could be useful for therapeutic purposes, such as restoring the mobility of the vocal cords in patients suffering from laryngeal paralysis.

[1]  R. James,et al.  The effect of physiological concentrations of caffeine on the power output of maximally and submaximally stimulated mouse EDL (fast) and soleus (slow) muscle. , 2012, Journal of applied physiology.

[2]  D. Kleinfeld,et al.  ReaChR: A red-shifted variant of channelrhodopsin enables deep transcranial optogenetic excitation , 2013, Nature Neuroscience.

[3]  Hiromu Yawo,et al.  Optically controlled contraction of photosensitive skeletal muscle cells , 2012, Biotechnology and bioengineering.

[4]  K. Deisseroth,et al.  Millisecond-timescale, genetically targeted optical control of neural activity , 2005, Nature Neuroscience.

[5]  T. McCulloch,et al.  Otolaryngology head and neck surgery: An integrative view of the larynx , 2011, Head & neck.

[6]  O. Delbono,et al.  Age-dependent fatigue in single intact fast- and slow fibers from mouse EDL and soleus skeletal muscles , 2001, Mechanisms of Ageing and Development.

[7]  Murtaza Z Mogri,et al.  Targeting and Readout Strategies for Fast Optical Neural Control In Vitro and In Vivo , 2007, The Journal of Neuroscience.

[8]  A. Pascual-Font,et al.  Functional role of human laryngeal nerve connections , 2011, The Laryngoscope.

[9]  R. Mainthia,et al.  Bilateral motion restored to the paralyzed canine larynx with implantable stimulator , 2010, The Laryngoscope.

[10]  M. Remacle,et al.  Subtotal Carbon Dioxide Laser Arytenoidectomy for the Treatment of Bilateral Vocal Fold Immobility: Long-Term Results , 2005, The Annals of otology, rhinology, and laryngology.

[11]  Elmar Brähler,et al.  Social withdrawal after laryngectomy , 2010, European Archives of Oto-Rhino-Laryngology.

[12]  Aristides B. Arrenberg,et al.  Optogenetic Control of Cardiac Function , 2010, Science.

[13]  K. L. Montgomery,et al.  Optogenetic Control of Targeted Peripheral Axons in Freely Moving Animals , 2013, PloS one.

[14]  B. Byrne,et al.  Sustained alpha‐sarcoglycan gene expression after gene transfer in limb‐girdle muscular dystrophy, type 2D , 2010, Annals of neurology.

[15]  R. Andreatta,et al.  Establishing a new animal model for the study of laryngeal biology and disease: an anatomic study of the mouse larynx. , 2009, Journal of speech, language, and hearing research : JSLHR.

[16]  Linda Greensmith,et al.  Optical Control of Muscle Function by Transplantation of Stem Cell–Derived Motor Neurons in Mice , 2014, Science.

[17]  J. Netterville,et al.  Reanimation of the Paralyzed Human Larynx With an Implantable Electrical Stimulation Device , 2003, The Laryngoscope.

[18]  Samad EJ Golzari,et al.  Vocal Cord Paralysis and its Etiologies: A Prospective Study , 2014, Journal of cardiovascular and thoracic research.

[19]  E. Bamberg,et al.  Channelrhodopsin-2, a directly light-gated cation-selective membrane channel , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  E. Bamberg,et al.  Light Activation of Channelrhodopsin-2 in Excitable Cells of Caenorhabditis elegans Triggers Rapid Behavioral Responses , 2005, Current Biology.

[21]  K. Deisseroth,et al.  Ultrafast optogenetic control , 2010, Nature Neuroscience.

[22]  T. Bruegmann,et al.  Optogenetic control of heart muscle in vitro and in vivo , 2010, Nature Methods.

[23]  Stefan R. Pulver,et al.  Independent Optical Excitation of Distinct Neural Populations , 2014, Nature Methods.

[24]  J. Grieger,et al.  Phase 1 gene therapy for Duchenne muscular dystrophy using a translational optimized AAV vector. , 2012, Molecular therapy : the journal of the American Society of Gene Therapy.

[25]  Xiao Xiao,et al.  Adeno-associated virus 9 mediated FKRP gene therapy restores functional glycosylation of α-dystroglycan and improves muscle functions. , 2013, Molecular therapy : the journal of the American Society of Gene Therapy.

[26]  K. Deisseroth,et al.  Orderly recruitment of motor units under optical control in vivo , 2010, Nature Medicine.

[27]  A. Rizzino,et al.  Temporally and spatially controlled expression of transgenes in embryonic and adult tissues , 2009, Transgenic Research.

[28]  B. Allard,et al.  Major contribution of sarcoplasmic reticulum Ca2+ depletion during long-lasting activation of skeletal muscle , 2013, The Journal of general physiology.

[29]  S. Tapscott,et al.  Immunity and AAV-Mediated Gene Therapy for Muscular Dystrophies in Large Animal Models and Human Trials , 2011, Front. Microbio..