1. A TAT-modified fusion protein efficiently penetrates mouse hypoglossal nuclei from transduced ependyma.
- Author
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Alisky JM, Xia H, and Davidson BL
- Subjects
- Adenoviridae genetics, Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis metabolism, Amyotrophic Lateral Sclerosis therapy, Animals, Area Postrema cytology, Area Postrema metabolism, Area Postrema virology, Ependyma cytology, Ependyma metabolism, Ependyma virology, Fourth Ventricle cytology, Fourth Ventricle metabolism, Fourth Ventricle virology, Gene Products, tat genetics, Gene Products, tat therapeutic use, Genetic Therapy methods, Genetic Vectors therapeutic use, Glucuronidase genetics, Hypoglossal Nerve cytology, Hypoglossal Nerve virology, Injections, Intraventricular, Medulla Oblongata cytology, Medulla Oblongata metabolism, Mice, Mice, Inbred C57BL, Mice, Transgenic, Motor Neurons cytology, Motor Neurons metabolism, Protein Structure, Tertiary genetics, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins therapeutic use, Transduction, Genetic trends, Transfection trends, Gene Products, tat metabolism, Genetic Vectors genetics, Hypoglossal Nerve metabolism, Recombinant Fusion Proteins metabolism, Transduction, Genetic methods, Transfection methods
- Abstract
Future gene therapy for brainstem variant amyotrophic lateral sclerosis may be technically difficult if gene therapy vectors are injected near vital cardiorespiratory centers or if large portions of the tongue and pharyngeal muscles must be peripherally injected for retrograde transport of vectors to motor neurons. In this study we show that it is possible to deliver recombinant proteins to the hypoglossal nuclei without brainstem or muscle injections, by taking advantage of enhanced uptake of fusion proteins containing the protein transduction domain from the human immunodeficiency virus TAT protein. Adenoviral vectors expressing either TAT-modified or native beta-glucuronidase (beta-gluc) were injected into the lateral cerebral ventricles of mice, transducing ventricular epithelium down to the level of the obex in the brainstem. There was significant uptake into the hypoglossal nuclei of TAT-modified but not native beta-glucuronidase. The TAT-modified beta-gluc appeared to encompass half or more of the hypoglossal nuclei as visualized by retrograde labeling with cholera toxin subunit b in adjacent sections. TAT-modification of gene products may allow a relatively non-invasive approach to brainstem gene therapy via cerebroventricular injection.
- Published
- 2006
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