Limits...
Protection by the NDI1 gene against neurodegeneration in a rotenone rat model of Parkinson's disease.

Marella M, Seo BB, Nakamaru-Ogiso E, Greenamyre JT, Matsuno-Yagi A, Yagi T - PLoS ONE (2008)

Bottom Line: It is widely recognized that mitochondrial dysfunction, most notably defects in the NADH-quinone oxidoreductase (complex I), is closely related to the etiology of sporadic Parkinson's disease (PD).A single, unilateral injection of recombinant adeno-associated virus carrying the NDI1 gene into the vicinity of the substantia nigra resulted in expression of the Ndi1 protein in the entire substantia nigra of that side.The present study shows, for the first time, the powerful neuroprotective effect offered by the Ndi1 enzyme in a rotenone rat model of PD.

View Article: PubMed Central - PubMed

Affiliation: Division of Biochemistry, Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, United States of America.

ABSTRACT
It is widely recognized that mitochondrial dysfunction, most notably defects in the NADH-quinone oxidoreductase (complex I), is closely related to the etiology of sporadic Parkinson's disease (PD). In fact, rotenone, a complex I inhibitor, has been used for establishing PD models both in vitro and in vivo. A rat model with chronic rotenone exposure seems to reproduce pathophysiological conditions of PD more closely than acute mouse models as manifested by neuronal cell death in the substantia nigra and Lewy body-like cytosolic aggregations. Using the rotenone rat model, we investigated the protective effects of alternative NADH dehydrogenase (Ndi1) which we previously demonstrated to act as a replacement for complex I both in vitro and in vivo. A single, unilateral injection of recombinant adeno-associated virus carrying the NDI1 gene into the vicinity of the substantia nigra resulted in expression of the Ndi1 protein in the entire substantia nigra of that side. It was clear that the introduction of the Ndi1 protein in the substantia nigra rendered resistance to the deleterious effects caused by rotenone exposure as assessed by the levels of tyrosine hydroxylase and dopamine. The presence of the Ndi1 protein also prevented cell death and oxidative damage to DNA in dopaminergic neurons observed in rotenone-treated rats. Unilateral protection also led to uni-directional rotation of the rotenone-exposed rats in the behavioral test. The present study shows, for the first time, the powerful neuroprotective effect offered by the Ndi1 enzyme in a rotenone rat model of PD.

Show MeSH

Related in: MedlinePlus

Prevention of rotenone-induced neuronal cell death and oxidative damage by expression of the Ndi1 protein.Rats were treated with the NDI1 gene and rotenone and brain sections were prepared and processed for immunohistochemical staining as described in Figure 2. A: Sections were subjected to Nissl staining and representative images are displayed (upper panel). The number of viable neurons was compared between the left and right SN by counting Nissl-positive cells in a given area of the SN (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (5 animals, 12 sections) and NDI1+rotenone (9 animals, 9 sections). In each group, images were collected from the sections with the matching anterior-posterior position and, when multiple sections were used in a given animal, they were separated by at least 30 µm to eliminate possible span of neuron cell bodies over multiple sections. B: Sections were stained for 8-oxo-dG to evaluate oxidative damage to DNA (upper panel). The SN neurons that are positively stained were counted (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (3 animals, 5 sections) and NDI1+rotenone (3 animals, 6 sections). Selection of the sections was done as described in A. Statistic analysis was done using student T-test. Error bars represent standard deviation.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2175531&req=5

pone-0001433-g003: Prevention of rotenone-induced neuronal cell death and oxidative damage by expression of the Ndi1 protein.Rats were treated with the NDI1 gene and rotenone and brain sections were prepared and processed for immunohistochemical staining as described in Figure 2. A: Sections were subjected to Nissl staining and representative images are displayed (upper panel). The number of viable neurons was compared between the left and right SN by counting Nissl-positive cells in a given area of the SN (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (5 animals, 12 sections) and NDI1+rotenone (9 animals, 9 sections). In each group, images were collected from the sections with the matching anterior-posterior position and, when multiple sections were used in a given animal, they were separated by at least 30 µm to eliminate possible span of neuron cell bodies over multiple sections. B: Sections were stained for 8-oxo-dG to evaluate oxidative damage to DNA (upper panel). The SN neurons that are positively stained were counted (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (3 animals, 5 sections) and NDI1+rotenone (3 animals, 6 sections). Selection of the sections was done as described in A. Statistic analysis was done using student T-test. Error bars represent standard deviation.

Mentions: As described above, rotenone exposure resulted in the appearance of cytoplasmic aggregations. This Lewy-body like structure seems to form less frequently, on the ipsilateral side, in the group of rats that were injected with the NDI1 gene. However, the total number of such inclusions was too small to obtain statistically reliable data. Neuronal cell death is another key observation in the rotenone rat model of PD. Approximately 50% of the SN neurons died in our rotenone model (Figure 3A). Comparison of the number of SN neurons between the injection side and the opposite side clearly showed that the Ndi1 protein expression prevented neuronal cell death in rotenone-treated animals. The preferential death of the nigrostriatal neurons under rotenone exposure may result from the synergic effect of ROS production by complex I inhibition and the presence of dopamine by way of production of oxyradicals and reactive intermediates [30], [31]. Our earlier experiments using rat PC12 cells demonstrated that the Ndi1 protein impedes the production of ROS triggered by complex I inhibition presumably by taking over electron transfer activity from endogenous complex I [17], [29], [32]. To examine whether the Ndi1 protein can exert the same ROS-reducing effect in vivo, we assessed the extent of oxidative DNA modifications in the SN neurons. Again, the data from the Ndi1-expressing SN exhibited almost no effect by rotenone whereas the opposite side underwent the same degree of damage as non-Ndi1 controls (Figure 3B).


Protection by the NDI1 gene against neurodegeneration in a rotenone rat model of Parkinson's disease.

Marella M, Seo BB, Nakamaru-Ogiso E, Greenamyre JT, Matsuno-Yagi A, Yagi T - PLoS ONE (2008)

Prevention of rotenone-induced neuronal cell death and oxidative damage by expression of the Ndi1 protein.Rats were treated with the NDI1 gene and rotenone and brain sections were prepared and processed for immunohistochemical staining as described in Figure 2. A: Sections were subjected to Nissl staining and representative images are displayed (upper panel). The number of viable neurons was compared between the left and right SN by counting Nissl-positive cells in a given area of the SN (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (5 animals, 12 sections) and NDI1+rotenone (9 animals, 9 sections). In each group, images were collected from the sections with the matching anterior-posterior position and, when multiple sections were used in a given animal, they were separated by at least 30 µm to eliminate possible span of neuron cell bodies over multiple sections. B: Sections were stained for 8-oxo-dG to evaluate oxidative damage to DNA (upper panel). The SN neurons that are positively stained were counted (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (3 animals, 5 sections) and NDI1+rotenone (3 animals, 6 sections). Selection of the sections was done as described in A. Statistic analysis was done using student T-test. Error bars represent standard deviation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2175531&req=5

pone-0001433-g003: Prevention of rotenone-induced neuronal cell death and oxidative damage by expression of the Ndi1 protein.Rats were treated with the NDI1 gene and rotenone and brain sections were prepared and processed for immunohistochemical staining as described in Figure 2. A: Sections were subjected to Nissl staining and representative images are displayed (upper panel). The number of viable neurons was compared between the left and right SN by counting Nissl-positive cells in a given area of the SN (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (5 animals, 12 sections) and NDI1+rotenone (9 animals, 9 sections). In each group, images were collected from the sections with the matching anterior-posterior position and, when multiple sections were used in a given animal, they were separated by at least 30 µm to eliminate possible span of neuron cell bodies over multiple sections. B: Sections were stained for 8-oxo-dG to evaluate oxidative damage to DNA (upper panel). The SN neurons that are positively stained were counted (lower panel). The number of sections used for analysis was; control (2 animals, 3 sections), control+rotenone (3 animals, 5 sections) and NDI1+rotenone (3 animals, 6 sections). Selection of the sections was done as described in A. Statistic analysis was done using student T-test. Error bars represent standard deviation.
Mentions: As described above, rotenone exposure resulted in the appearance of cytoplasmic aggregations. This Lewy-body like structure seems to form less frequently, on the ipsilateral side, in the group of rats that were injected with the NDI1 gene. However, the total number of such inclusions was too small to obtain statistically reliable data. Neuronal cell death is another key observation in the rotenone rat model of PD. Approximately 50% of the SN neurons died in our rotenone model (Figure 3A). Comparison of the number of SN neurons between the injection side and the opposite side clearly showed that the Ndi1 protein expression prevented neuronal cell death in rotenone-treated animals. The preferential death of the nigrostriatal neurons under rotenone exposure may result from the synergic effect of ROS production by complex I inhibition and the presence of dopamine by way of production of oxyradicals and reactive intermediates [30], [31]. Our earlier experiments using rat PC12 cells demonstrated that the Ndi1 protein impedes the production of ROS triggered by complex I inhibition presumably by taking over electron transfer activity from endogenous complex I [17], [29], [32]. To examine whether the Ndi1 protein can exert the same ROS-reducing effect in vivo, we assessed the extent of oxidative DNA modifications in the SN neurons. Again, the data from the Ndi1-expressing SN exhibited almost no effect by rotenone whereas the opposite side underwent the same degree of damage as non-Ndi1 controls (Figure 3B).

Bottom Line: It is widely recognized that mitochondrial dysfunction, most notably defects in the NADH-quinone oxidoreductase (complex I), is closely related to the etiology of sporadic Parkinson's disease (PD).A single, unilateral injection of recombinant adeno-associated virus carrying the NDI1 gene into the vicinity of the substantia nigra resulted in expression of the Ndi1 protein in the entire substantia nigra of that side.The present study shows, for the first time, the powerful neuroprotective effect offered by the Ndi1 enzyme in a rotenone rat model of PD.

View Article: PubMed Central - PubMed

Affiliation: Division of Biochemistry, Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, United States of America.

ABSTRACT
It is widely recognized that mitochondrial dysfunction, most notably defects in the NADH-quinone oxidoreductase (complex I), is closely related to the etiology of sporadic Parkinson's disease (PD). In fact, rotenone, a complex I inhibitor, has been used for establishing PD models both in vitro and in vivo. A rat model with chronic rotenone exposure seems to reproduce pathophysiological conditions of PD more closely than acute mouse models as manifested by neuronal cell death in the substantia nigra and Lewy body-like cytosolic aggregations. Using the rotenone rat model, we investigated the protective effects of alternative NADH dehydrogenase (Ndi1) which we previously demonstrated to act as a replacement for complex I both in vitro and in vivo. A single, unilateral injection of recombinant adeno-associated virus carrying the NDI1 gene into the vicinity of the substantia nigra resulted in expression of the Ndi1 protein in the entire substantia nigra of that side. It was clear that the introduction of the Ndi1 protein in the substantia nigra rendered resistance to the deleterious effects caused by rotenone exposure as assessed by the levels of tyrosine hydroxylase and dopamine. The presence of the Ndi1 protein also prevented cell death and oxidative damage to DNA in dopaminergic neurons observed in rotenone-treated rats. Unilateral protection also led to uni-directional rotation of the rotenone-exposed rats in the behavioral test. The present study shows, for the first time, the powerful neuroprotective effect offered by the Ndi1 enzyme in a rotenone rat model of PD.

Show MeSH
Related in: MedlinePlus