Transduction of motor neurons and muscle fibers by intramuscular injection of HI

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J Neurosci Methods. 2006 May 23

Transduction of motor neurons and muscle fibers by intramuscular

injection of HIV-1-based vectors pseudotyped with select rabies virus

glycoproteins.

Mentis GZ, Gravell M, Hamilton R, Shneider NA, O'donovan MJ, Schubert

M.

Developmental Neurobiology Section, Basic Neuroscience Program,

National Institutes of Neurological Disorders and Stroke, National

Institutes of Health, 35 Convent Drive, Building 35, Rm. 3BC1014,

Bethesda, MD 20892-3700, USA.

For studies of motor neuron function or for therapeutic purposes,

novel pseudotype HIV-1-based vectors were developed that are capable

of expressing transgenes in motor neurons following injection into

mouse hind limb muscles. To specifically target motor neurons,

glycoproteins from two rabies virus (RV) isolates, the mouse-brain

adapted challenge virus 24 (CVS-24) variants, CVS-N2c and CVS-B2c

were evaluated for pseudotype formation with an HIV-1-based vector.

Both RV glycoproteins incorporated into vector envelopes, and both

pseudotypes yielded high titers with Hek293T and cortical plate

neuron cultures.

Increased neuronotropism by the CVS-N2c pseudotype was not observed,

suggesting that vector tropism is not solely determined by the

fusogenic viral glycoprotein.

Vector injection into hind limb muscles resulted in EYFP reporter

gene expression in the injected muscle fibers and in spinal cord

motor neurons innervating the same muscle, indicating retrograde

vector transport. Intramuscular vector injections into the soleus and

tibialis anterior muscles transduced 26% and 16% of all motor neurons

in each motor nucleus, respectively.

These transduction efficiencies may allow novel approaches to

functional studies of the motor system and the treatment of

neuromuscular disease.