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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

Neuron-specific transduction in the cerebellar cortex and deep nuclei resulting from all three routes of viral administration. Sagittal (left images) and transverse (middle and right images) sections of the cerebellum immunostained for GFP and Nissl substance (left and right images) or calbindin D-28K (calbindin) (middle images). (a–d) Sections from the mice that received DC injections. (e–h) Sections from the mice that received IT injections. (i–l) Sections from the mice that received IV injections. Arrowheads show interneurons, which were transduced only by DC and IT injections. Thick and thin arrows indicate transduced PCs and projection neurons in the DCN, respectively. Interneurons in the DCN, which are labeled by asterisks, were not transduced by any of the three administration routes. (m) Quantitative analysis of transduction efficacy of PCs by three injection routes. Graph shows the % ratio of the transduced PCs to all PCs were determined in four cerebellar groups as shown in Figure 3j. Each value was obtained from three slices/mouse using three mice in each viral injection group. Scale bars, 50 µm, except for panels (d,h,l) which are 20 µm. Asterisks indicate statistically significant differences determined by one-way analysis of variance followed by Tukey’s post hoc test, **P < 0.01.
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fig4: Neuron-specific transduction in the cerebellar cortex and deep nuclei resulting from all three routes of viral administration. Sagittal (left images) and transverse (middle and right images) sections of the cerebellum immunostained for GFP and Nissl substance (left and right images) or calbindin D-28K (calbindin) (middle images). (a–d) Sections from the mice that received DC injections. (e–h) Sections from the mice that received IT injections. (i–l) Sections from the mice that received IV injections. Arrowheads show interneurons, which were transduced only by DC and IT injections. Thick and thin arrows indicate transduced PCs and projection neurons in the DCN, respectively. Interneurons in the DCN, which are labeled by asterisks, were not transduced by any of the three administration routes. (m) Quantitative analysis of transduction efficacy of PCs by three injection routes. Graph shows the % ratio of the transduced PCs to all PCs were determined in four cerebellar groups as shown in Figure 3j. Each value was obtained from three slices/mouse using three mice in each viral injection group. Scale bars, 50 µm, except for panels (d,h,l) which are 20 µm. Asterisks indicate statistically significant differences determined by one-way analysis of variance followed by Tukey’s post hoc test, **P < 0.01.

Mentions: We next examined the cerebellar cell types transduced by the three different routes of viral administration. PCs, the sole output neurons from the cerebellar cortex, have large somata with well-differentiated dendrites and play critical roles in the processing of motor coordination. To examine the transduction of PCs, the cerebellar sections were double immunostained for GFP and Nissl substance or calbindin D-28K (calbindin), a PC marker. We observed that all three routes of vector administration produced an efficient transduction of PCs (Figure 4a,e,i, thick arrows), and this finding was confirmed by immunolabeling PCs with anti-calbindin antibody (Figure 4b,f,j, thick arrows).


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)

Neuron-specific transduction in the cerebellar cortex and deep nuclei resulting from all three routes of viral administration. Sagittal (left images) and transverse (middle and right images) sections of the cerebellum immunostained for GFP and Nissl substance (left and right images) or calbindin D-28K (calbindin) (middle images). (a–d) Sections from the mice that received DC injections. (e–h) Sections from the mice that received IT injections. (i–l) Sections from the mice that received IV injections. Arrowheads show interneurons, which were transduced only by DC and IT injections. Thick and thin arrows indicate transduced PCs and projection neurons in the DCN, respectively. Interneurons in the DCN, which are labeled by asterisks, were not transduced by any of the three administration routes. (m) Quantitative analysis of transduction efficacy of PCs by three injection routes. Graph shows the % ratio of the transduced PCs to all PCs were determined in four cerebellar groups as shown in Figure 3j. Each value was obtained from three slices/mouse using three mice in each viral injection group. Scale bars, 50 µm, except for panels (d,h,l) which are 20 µm. Asterisks indicate statistically significant differences determined by one-way analysis of variance followed by Tukey’s post hoc test, **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Neuron-specific transduction in the cerebellar cortex and deep nuclei resulting from all three routes of viral administration. Sagittal (left images) and transverse (middle and right images) sections of the cerebellum immunostained for GFP and Nissl substance (left and right images) or calbindin D-28K (calbindin) (middle images). (a–d) Sections from the mice that received DC injections. (e–h) Sections from the mice that received IT injections. (i–l) Sections from the mice that received IV injections. Arrowheads show interneurons, which were transduced only by DC and IT injections. Thick and thin arrows indicate transduced PCs and projection neurons in the DCN, respectively. Interneurons in the DCN, which are labeled by asterisks, were not transduced by any of the three administration routes. (m) Quantitative analysis of transduction efficacy of PCs by three injection routes. Graph shows the % ratio of the transduced PCs to all PCs were determined in four cerebellar groups as shown in Figure 3j. Each value was obtained from three slices/mouse using three mice in each viral injection group. Scale bars, 50 µm, except for panels (d,h,l) which are 20 µm. Asterisks indicate statistically significant differences determined by one-way analysis of variance followed by Tukey’s post hoc test, **P < 0.01.
Mentions: We next examined the cerebellar cell types transduced by the three different routes of viral administration. PCs, the sole output neurons from the cerebellar cortex, have large somata with well-differentiated dendrites and play critical roles in the processing of motor coordination. To examine the transduction of PCs, the cerebellar sections were double immunostained for GFP and Nissl substance or calbindin D-28K (calbindin), a PC marker. We observed that all three routes of vector administration produced an efficient transduction of PCs (Figure 4a,e,i, thick arrows), and this finding was confirmed by immunolabeling PCs with anti-calbindin antibody (Figure 4b,f,j, thick arrows).

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