Use of Self-Complementary Adeno-Associated Virus Serotype 2 as a Tracer for Labeling Axons: Implications for Axon Regeneration
Various types of tracers are available for use in axon regeneration, but they require an extra operational tracer injection, time-consuming immunohistochemical analysis and cause non-specific labeling. Considerable efforts over the past years have explored other methodologies, especially the use of...
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| Veröffentlicht in: | PloS one Jg. 9; H. 2; S. e87447 |
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| Abstract | Various types of tracers are available for use in axon regeneration, but they require an extra operational tracer injection, time-consuming immunohistochemical analysis and cause non-specific labeling. Considerable efforts over the past years have explored other methodologies, especially the use of viral vectors, to investigate axon regeneration after injury. Recent studies have demonstrated that self-complementary Adeno-Associated Virus (scAAV) induced a high transduction efficiency and faster expression of transgenes. Here, we describe for the first time the use of scAAV2-GFP to label long-projection axons in the corticospinal tract (CST), rubrospinal tract (RST) and the central axons of dorsal root ganglion (DRG) in the normal and lesioned animal models. We found that scAAV2-GFP could efficiently transduce neurons in the sensorimotor cortex, red nucleus and DRG. Strong GFP expression could be transported anterogradely along the axon to label the numerous axon fibers from CST, RST and central axons of DRG separately. Comparison of the scAAV2 vector with single-stranded (ss) AAV2 vector in co-labeled sections showed that the scAAV2 vector induced a faster and stronger transgene expression than the ssAAV2 vector in DRG neurons and their axons. In both spinal cord lesion and dorsal root crush injury models, scAAV-GFP could efficiently label the lesioned and regenerated axons around the lesion cavity and the dorsal root entry zone (DREZ) respectively. Further, scAAV2-GFP vector could be combined with traditional tracer to specifically label sensory and motor axons after spinal cord lesion. Thus, we show that using scAAV2-GFP as a tracer is a more effective and efficient way to study axon regeneration following injury. |
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| AbstractList | Various types of tracers are available for use in axon regeneration, but they require an extra operational tracer injection, time-consuming immunohistochemical analysis and cause non-specific labeling. Considerable efforts over the past years have explored other methodologies, especially the use of viral vectors, to investigate axon regeneration after injury. Recent studies have demonstrated that self-complementary Adeno-Associated Virus (scAAV) induced a high transduction efficiency and faster expression of transgenes. Here, we describe for the first time the use of scAAV2-GFP to label long-projection axons in the corticospinal tract (CST), rubrospinal tract (RST) and the central axons of dorsal root ganglion (DRG) in the normal and lesioned animal models. We found that scAAV2-GFP could efficiently transduce neurons in the sensorimotor cortex, red nucleus and DRG. Strong GFP expression could be transported anterogradely along the axon to label the numerous axon fibers from CST, RST and central axons of DRG separately. Comparison of the scAAV2 vector with single-stranded (ss) AAV2 vector in co-labeled sections showed that the scAAV2 vector induced a faster and stronger transgene expression than the ssAAV2 vector in DRG neurons and their axons. In both spinal cord lesion and dorsal root crush injury models, scAAV-GFP could efficiently label the lesioned and regenerated axons around the lesion cavity and the dorsal root entry zone (DREZ) respectively. Further, scAAV2-GFP vector could be combined with traditional tracer to specifically label sensory and motor axons after spinal cord lesion. Thus, we show that using scAAV2-GFP as a tracer is a more effective and efficient way to study axon regeneration following injury. Various types of tracers are available for use in axon regeneration, but they require an extra operational tracer injection, time-consuming immunohistochemical analysis and cause non-specific labeling. Considerable efforts over the past years have explored other methodologies, especially the use of viral vectors, to investigate axon regeneration after injury. Recent studies have demonstrated that self-complementary Adeno-Associated Virus (scAAV) induced a high transduction efficiency and faster expression of transgenes. Here, we describe for the first time the use of scAAV2-GFP to label long-projection axons in the corticospinal tract (CST), rubrospinal tract (RST) and the central axons of dorsal root ganglion (DRG) in the normal and lesioned animal models. We found that scAAV2-GFP could efficiently transduce neurons in the sensorimotor cortex, red nucleus and DRG. Strong GFP expression could be transported anterogradely along the axon to label the numerous axon fibers from CST, RST and central axons of DRG separately. Comparison of the scAAV2 vector with single-stranded (ss) AAV2 vector in co-labeled sections showed that the scAAV2 vector induced a faster and stronger transgene expression than the ssAAV2 vector in DRG neurons and their axons. In both spinal cord lesion and dorsal root crush injury models, scAAV-GFP could efficiently label the lesioned and regenerated axons around the lesion cavity and the dorsal root entry zone (DREZ) respectively. Further, scAAV2-GFP vector could be combined with traditional tracer to specifically label sensory and motor axons after spinal cord lesion. Thus, we show that using scAAV2-GFP as a tracer is a more effective and efficient way to study axon regeneration following injury.Various types of tracers are available for use in axon regeneration, but they require an extra operational tracer injection, time-consuming immunohistochemical analysis and cause non-specific labeling. Considerable efforts over the past years have explored other methodologies, especially the use of viral vectors, to investigate axon regeneration after injury. Recent studies have demonstrated that self-complementary Adeno-Associated Virus (scAAV) induced a high transduction efficiency and faster expression of transgenes. Here, we describe for the first time the use of scAAV2-GFP to label long-projection axons in the corticospinal tract (CST), rubrospinal tract (RST) and the central axons of dorsal root ganglion (DRG) in the normal and lesioned animal models. We found that scAAV2-GFP could efficiently transduce neurons in the sensorimotor cortex, red nucleus and DRG. Strong GFP expression could be transported anterogradely along the axon to label the numerous axon fibers from CST, RST and central axons of DRG separately. Comparison of the scAAV2 vector with single-stranded (ss) AAV2 vector in co-labeled sections showed that the scAAV2 vector induced a faster and stronger transgene expression than the ssAAV2 vector in DRG neurons and their axons. In both spinal cord lesion and dorsal root crush injury models, scAAV-GFP could efficiently label the lesioned and regenerated axons around the lesion cavity and the dorsal root entry zone (DREZ) respectively. Further, scAAV2-GFP vector could be combined with traditional tracer to specifically label sensory and motor axons after spinal cord lesion. Thus, we show that using scAAV2-GFP as a tracer is a more effective and efficient way to study axon regeneration following injury. |
| Audience | Academic |
| Author | Liu, Yingpeng Lin, Shen Tang, Xiaoqing Smith, George M. Keefe, Kathy |
| AuthorAffiliation | Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, United States of America Hertie Institute for Clinical Brain Research, University of Tuebingen., Germany |
| AuthorAffiliation_xml | – name: Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania, United States of America – name: Hertie Institute for Clinical Brain Research, University of Tuebingen., Germany |
| Author_xml | – sequence: 1 givenname: Yingpeng surname: Liu fullname: Liu, Yingpeng – sequence: 2 givenname: Kathy surname: Keefe fullname: Keefe, Kathy – sequence: 3 givenname: Xiaoqing surname: Tang fullname: Tang, Xiaoqing – sequence: 4 givenname: Shen surname: Lin fullname: Lin, Shen – sequence: 5 givenname: George M. surname: Smith fullname: Smith, George M. |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/24498323$$D View this record in MEDLINE/PubMed |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Competing Interests: The authors have declared that no competing interests exist. Conceived and designed the experiments: YL KK GMS. Performed the experiments: YL KK XT SL. Analyzed the data: YL KK XT GMS. Contributed reagents/materials/analysis tools: YL KK XT GMS. Wrote the paper: YL KK XT SL GMS. |
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| SubjectTerms | Adenoviruses Analysis Animal models Animals Axons Axons - metabolism Axons - physiology Biology Brain - metabolism Brain research Cortex (somatosensory) Deoxyribonucleic acid Dependovirus - genetics DNA Dorsal root ganglia Fibers Fibroblasts Ganglia, Spinal - metabolism Gene expression Gene therapy Genetic Vectors - genetics Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism HEK293 Cells Hospitals Humans Injuries Labeling Labelling Laboratory animals Lesions Luminescent Proteins - genetics Luminescent Proteins - metabolism Medicine Microscopy, Fluorescence Motor neurons Motor Neurons - metabolism Nerve Regeneration Neurons Pediatrics Plasmids Pyramidal tracts Rats Rats, Inbred F344 Red Fluorescent Protein Red nucleus Regeneration Rodents Rubrospinal tract Sensory neurons Sensory Receptor Cells - metabolism Somatosensory cortex Spinal cord Spinal Cord - metabolism Spinal cord injuries Spinal Nerve Roots - metabolism Tracers Tracers (Biology) Transgenes Vectors (Biology) Viruses |
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| Title | Use of Self-Complementary Adeno-Associated Virus Serotype 2 as a Tracer for Labeling Axons: Implications for Axon Regeneration |
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