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Distinct transduction profiles in the CNS via three injection routes of AAV9 and the application to generation of a neurodegenerative mouse model.

Huda F, Konno A, Matsuzaki Y, Goenawan H, Miyake K, Shimada T, Hirai H - Mol Ther Methods Clin Dev (2014)

Bottom Line: Using single-stranded adeno-associated virus serotype 9 (ssAAV9) vectors containing the neuron-specific synapsin-I promoter, we examined whether different administration routes (direct cerebellar cortical (DC), intrathecal (IT) and intravenous (IV) injections) could elicit specific transduction profiles in the CNS.In the cerebellar cortex, the DC and IT injection routes transduced all neuron types, whereas the IV injection route primarily transduced Purkinje cells.Thus, ssAAV9-mediated transduction areas, levels, and cell types change depending on the route of injection.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurophysiology, Gunma University Graduate School of Medicine , Maebashi, Japan ; Department of Physiology, Faculty of Medicine Universitas Padjadjaran , Bandung, Indonesia.

ABSTRACT
Using single-stranded adeno-associated virus serotype 9 (ssAAV9) vectors containing the neuron-specific synapsin-I promoter, we examined whether different administration routes (direct cerebellar cortical (DC), intrathecal (IT) and intravenous (IV) injections) could elicit specific transduction profiles in the CNS. The DC injection route robustly and exclusively transduced the whole cerebellum, whereas the IT injection route primarily transduced the cerebellar lobules 9 and 10 close to the injection site and the spinal cord. An IV injection in neonatal mice weakly and homogenously transduced broad CNS areas. In the cerebellar cortex, the DC and IT injection routes transduced all neuron types, whereas the IV injection route primarily transduced Purkinje cells. To verify the usefulness of this method, we generated a mouse model of spinocerebellar ataxia type 1 (SCA1). Mice that received a DC injection of the ssAAV9 vector expressing mutant ATXN1, a protein responsible for SCA1, showed the intranuclear aggregation of mutant ATXN1 in Purkinje cells, significant atrophy of the Purkinje cell dendrites and progressive motor deficits, which are characteristics of SCA1. Thus, ssAAV9-mediated transduction areas, levels, and cell types change depending on the route of injection. Moreover, this approach can be used for the generation of different mouse models of CNS/neurodegenerative diseases.

No MeSH data available.


Related in: MedlinePlus

Progressive behavioral impairment in mice that received DC injection of ssAAV9 vectors expressing ATXN1(Q141). Four-week-old C57BL/6 mice received an injection of phosphate-buffered saline (PBS) or ssAAV9 vectors expressing ATXN1(Q141) or GFP. (a,b) The motor coordination of the mice was tested with a rotating rod. Mice underwent four trials per day on both the accelerating (a) (4–40 rpm) rotarod and subsequently the constant (b) (20 rpm) rotarod immediately before and every week up to 8 weeks after the injection. The rotarod performances of the mice expressing mutant ATXN1 (n = 6 mice) were compared with those of the mice treated with PBS (n = 6) or the mice expressing GFP (n = 6). (c) Body weights of mice from the three experimental groups. Asterisks and daggers indicate statistically significant differences compared with the PBS-injected mice and the mice injected with ssAAV9 vectors expressing GFP, respectively, as determined by one-way analysis of variance followed by Tukey’s post hoc test, *P < 0.05, **P < 0.01, ***P < 0.001, ††P < 0.01, and †††P < 0.001.
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fig7: Progressive behavioral impairment in mice that received DC injection of ssAAV9 vectors expressing ATXN1(Q141). Four-week-old C57BL/6 mice received an injection of phosphate-buffered saline (PBS) or ssAAV9 vectors expressing ATXN1(Q141) or GFP. (a,b) The motor coordination of the mice was tested with a rotating rod. Mice underwent four trials per day on both the accelerating (a) (4–40 rpm) rotarod and subsequently the constant (b) (20 rpm) rotarod immediately before and every week up to 8 weeks after the injection. The rotarod performances of the mice expressing mutant ATXN1 (n = 6 mice) were compared with those of the mice treated with PBS (n = 6) or the mice expressing GFP (n = 6). (c) Body weights of mice from the three experimental groups. Asterisks and daggers indicate statistically significant differences compared with the PBS-injected mice and the mice injected with ssAAV9 vectors expressing GFP, respectively, as determined by one-way analysis of variance followed by Tukey’s post hoc test, *P < 0.05, **P < 0.01, ***P < 0.001, ††P < 0.01, and †††P < 0.001.

Mentions: To prove the utility of the present method for the production of SCA animal models, we used the DC injection route because the DC injection of ssAAV9 vectors elicited efficient and robust transgene expression throughout the whole cerebellum (Figures 3i and 4m). Four-week-old C57BL/6 mice that showed approximately similar rotarod performances received a DC injection of ssAAV9 vectors expressing GFP or human influenza hemagglutinin (HA)-tagged ATXN1 with an abnormally expanded (141 repeats) polyglutamine stretch under the control of the enhanced synapsin I promoter (ssAAV9-Syn1minCMV-GFP-WPRE or ssAAV9-Syn1minCMV-HA-ATXN1(Q141)-WPRE). Sham-operated mice received a DC injection of a similar volume (10 µl) of phosphate-buffered saline (PBS). Three mouse groups (six mice each), i.e., a PBS-injected group, a GFP-expressing group and a mutant ATXN1-expressing group, showed no statistically significant difference in rotarod performance until 4 weeks after the injection. However, the mutant ATXN1-expressing, but not the GFP-expressing, mice showed a significantly poorer performance than the control PBS-injected mice from 5 weeks after the injection (Figure 7a,b). The impaired performance of ATXN1-expressing mice became more evident thereafter and continued during the observation period (at least, up to 11 weeks after the injection). The weight of the mice did not change significantly among the three groups during the observation period (Figure 7c), suggesting that the poor rotarod performance was not due to the maldevelopment of ATXN1-expressing mice.


Distinct transduction profiles in the CNS via three injection routes of AAV9 and the application to generation of a neurodegenerative mouse model.

Huda F, Konno A, Matsuzaki Y, Goenawan H, Miyake K, Shimada T, Hirai H - Mol Ther Methods Clin Dev (2014)

Progressive behavioral impairment in mice that received DC injection of ssAAV9 vectors expressing ATXN1(Q141). Four-week-old C57BL/6 mice received an injection of phosphate-buffered saline (PBS) or ssAAV9 vectors expressing ATXN1(Q141) or GFP. (a,b) The motor coordination of the mice was tested with a rotating rod. Mice underwent four trials per day on both the accelerating (a) (4–40 rpm) rotarod and subsequently the constant (b) (20 rpm) rotarod immediately before and every week up to 8 weeks after the injection. The rotarod performances of the mice expressing mutant ATXN1 (n = 6 mice) were compared with those of the mice treated with PBS (n = 6) or the mice expressing GFP (n = 6). (c) Body weights of mice from the three experimental groups. Asterisks and daggers indicate statistically significant differences compared with the PBS-injected mice and the mice injected with ssAAV9 vectors expressing GFP, respectively, as determined by one-way analysis of variance followed by Tukey’s post hoc test, *P < 0.05, **P < 0.01, ***P < 0.001, ††P < 0.01, and †††P < 0.001.
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Related In: Results  -  Collection

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fig7: Progressive behavioral impairment in mice that received DC injection of ssAAV9 vectors expressing ATXN1(Q141). Four-week-old C57BL/6 mice received an injection of phosphate-buffered saline (PBS) or ssAAV9 vectors expressing ATXN1(Q141) or GFP. (a,b) The motor coordination of the mice was tested with a rotating rod. Mice underwent four trials per day on both the accelerating (a) (4–40 rpm) rotarod and subsequently the constant (b) (20 rpm) rotarod immediately before and every week up to 8 weeks after the injection. The rotarod performances of the mice expressing mutant ATXN1 (n = 6 mice) were compared with those of the mice treated with PBS (n = 6) or the mice expressing GFP (n = 6). (c) Body weights of mice from the three experimental groups. Asterisks and daggers indicate statistically significant differences compared with the PBS-injected mice and the mice injected with ssAAV9 vectors expressing GFP, respectively, as determined by one-way analysis of variance followed by Tukey’s post hoc test, *P < 0.05, **P < 0.01, ***P < 0.001, ††P < 0.01, and †††P < 0.001.
Mentions: To prove the utility of the present method for the production of SCA animal models, we used the DC injection route because the DC injection of ssAAV9 vectors elicited efficient and robust transgene expression throughout the whole cerebellum (Figures 3i and 4m). Four-week-old C57BL/6 mice that showed approximately similar rotarod performances received a DC injection of ssAAV9 vectors expressing GFP or human influenza hemagglutinin (HA)-tagged ATXN1 with an abnormally expanded (141 repeats) polyglutamine stretch under the control of the enhanced synapsin I promoter (ssAAV9-Syn1minCMV-GFP-WPRE or ssAAV9-Syn1minCMV-HA-ATXN1(Q141)-WPRE). Sham-operated mice received a DC injection of a similar volume (10 µl) of phosphate-buffered saline (PBS). Three mouse groups (six mice each), i.e., a PBS-injected group, a GFP-expressing group and a mutant ATXN1-expressing group, showed no statistically significant difference in rotarod performance until 4 weeks after the injection. However, the mutant ATXN1-expressing, but not the GFP-expressing, mice showed a significantly poorer performance than the control PBS-injected mice from 5 weeks after the injection (Figure 7a,b). The impaired performance of ATXN1-expressing mice became more evident thereafter and continued during the observation period (at least, up to 11 weeks after the injection). The weight of the mice did not change significantly among the three groups during the observation period (Figure 7c), suggesting that the poor rotarod performance was not due to the maldevelopment of ATXN1-expressing mice.

Bottom Line: Using single-stranded adeno-associated virus serotype 9 (ssAAV9) vectors containing the neuron-specific synapsin-I promoter, we examined whether different administration routes (direct cerebellar cortical (DC), intrathecal (IT) and intravenous (IV) injections) could elicit specific transduction profiles in the CNS.In the cerebellar cortex, the DC and IT injection routes transduced all neuron types, whereas the IV injection route primarily transduced Purkinje cells.Thus, ssAAV9-mediated transduction areas, levels, and cell types change depending on the route of injection.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurophysiology, Gunma University Graduate School of Medicine , Maebashi, Japan ; Department of Physiology, Faculty of Medicine Universitas Padjadjaran , Bandung, Indonesia.

ABSTRACT
Using single-stranded adeno-associated virus serotype 9 (ssAAV9) vectors containing the neuron-specific synapsin-I promoter, we examined whether different administration routes (direct cerebellar cortical (DC), intrathecal (IT) and intravenous (IV) injections) could elicit specific transduction profiles in the CNS. The DC injection route robustly and exclusively transduced the whole cerebellum, whereas the IT injection route primarily transduced the cerebellar lobules 9 and 10 close to the injection site and the spinal cord. An IV injection in neonatal mice weakly and homogenously transduced broad CNS areas. In the cerebellar cortex, the DC and IT injection routes transduced all neuron types, whereas the IV injection route primarily transduced Purkinje cells. To verify the usefulness of this method, we generated a mouse model of spinocerebellar ataxia type 1 (SCA1). Mice that received a DC injection of the ssAAV9 vector expressing mutant ATXN1, a protein responsible for SCA1, showed the intranuclear aggregation of mutant ATXN1 in Purkinje cells, significant atrophy of the Purkinje cell dendrites and progressive motor deficits, which are characteristics of SCA1. Thus, ssAAV9-mediated transduction areas, levels, and cell types change depending on the route of injection. Moreover, this approach can be used for the generation of different mouse models of CNS/neurodegenerative diseases.

No MeSH data available.


Related in: MedlinePlus