Biomedical subjects

Kenji Yoshimi

Publications and source records attributed to Kenji Yoshimi.

8 recordsLinked to original sources

PubMed2006

Dopaminergic neuronal loss in transgenic mice expressing the Parkinson's disease-associated UCH-L1 I93M mutant.

The I93M mutation in ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) was reported in one German family with autosomal dominant Parkinson's disease (PD). The causative role of the mutation has, however, been questioned. We generated transgenic (Tg) mice carrying human UCHL1 under control of the PDGF-B promoter; two independent lines were generated with the I93M mutation (a high- and low-expressing line) and one line with wild-type human UCH-L1. We found a significant reduction in the dopaminergic neurons in the substantia nigra and the dopamine content in the striatum in the high-expressing I93M Tg mice as compared with non-Tg mice at 20 weeks of age. Although these changes were absent in the low-expressing I93M Tg mice, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment profoundly reduced dopaminergic neurons in this line as compared with wild-type Tg or non-Tg mice. Abnormal neuropathologies were also observed, such as silver staining-positive argyrophilic grains in the perikarya of degenerating dopaminergic neurons, in I93M Tg mice. The midbrains of I93M Tg mice contained increased amounts of insoluble UCH-L1 as compared with those of non-Tg mice, perhaps resulting in a toxic gain of function. Collectively, our data represent in vivo evidence that expression of UCHL1(I93M) leads to the degeneration of dopaminergic neurons.

Animals↗

PubMed2006

Statistical parametric mapping of immunopositive cell density.

We developed a new method for comparing immunopositive cell densities across groups of animals and creating statistical parametric maps on standardized sections. As an example, we compared Iba-1 (microglial marker) positive cell densities in rats with (n=6) and without (n=6) unilateral injection of 1-methyl-4-phenylpyridinium salt (MPP+). Immunopositive cells were automatically counted in each animal over a coronal section in the midbrain (bregma -5.9 mm) and a positive cell density map was created for each animal. After the positive cell density map was normalized to a template section from an atlas, positive cell densities of the two groups were compared in each pixel over the section and a statistical parameter (p-value from t-test) was mapped on each pixel. We were able to detect significant increases of microglias in the side of MPP+ injection not only in the substantia nigra pars compacta but also in adjacent white matter. We also applied the same analysis to tyrosine hydroxylase stained sections and detected significant decreases of dopamine neurons in the side of MPP+ injection. The new method was proven to be useful for detecting significant changes of cell densities over the entire area of immunostained sections.

1-Methyl-4-phenylpyridinium↗

PubMed2006

Genetic vitamin E deficiency does not affect MPTP susceptibility in the mouse brain.

Oxidative stress is involved in the degeneration of the nigrostriatal dopaminergic system in Parkinson's disease (PD). Vitamin E (alpha-tocopherol) is a potent antioxidant in the cell membrane that can trap free radicals and prohibit lipid peroxidation. The retention and secretion of vitamin E are regulated by alpha-tocopherol transfer protein (TTP) in the brain and liver. Dysfunction of TTP results in systemic deficiency of vitamin E in humans and mice, and increased oxidative stress in mouse brain. In this study, we investigated the effect of vitamin E deficiency in PD development by generating an 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD using TTP knockout (TTP-/-) mice. Vitamin E concentration in the brains of TTP+/- mice was half that in TTP+/+ mice, and in TTP-/- mice, was undetectable. MPTP treatment tended to decrease striatal dopamine, but the effect was comparable and not significant in any of the three genotypes. Furthermore, the extent of loss of dopaminergic cell bodies in the substantia nigra did not differ among the groups. One the other hand, oral administration of vitamin E resulted in the partial protection of striatal dopaminergic terminals against MPTP toxicity. Our results suggest that vitamin E does not play a major protective role in MPTP-induced nigrostriatal dopaminergic neurodegeneration in the brain.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed2005

Possibility for neurogenesis in substantia nigra of parkinsonian brain.

Recent studies of enhanced hippocampal neurogenesis by antidepressants suggest enhancement of neurogenesis is a potentially effective therapy in neurodegenerative diseases. In this study, we evaluated nigral neurogenesis in animals and autopsy brains including patients with Parkinson's disease (PD). First, proliferating cells in substantia nigra were labeled with retroviral transduction of green fluorescent protein, which is an efficient method to label neuronal stem cells. Subsequent differentiation of labeled cells was followed; many transduced cells became microglia, but no differentiation into tyrosine hydroxylase-positive neurons was detected at 4 weeks after injection, in both intact rodents and those treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Second, polysialic acid (PSA)-like immunoreactivity, indicative of newly differentiated neurons, was detected in the substantia nigra of rodent, primate, and human midbrains. A large number of PSA-positive cells were detected in the substantia nigra pars reticulata of some patients with PD. In rats and a macaque monkey, the dopamine-depleted hemispheres showed more PSA staining than the intact side. A small number of tyrosine hydroxylase-positive cells were PSA-positive. Our results suggest enhanced neural reconstruction in PD, which may be important in the design of new therapies against the progression of PD.

Aged↗

PubMed2004

Caspase-11 mediates inflammatory dopaminergic cell death in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease.

The present study was designed to elucidate the inflammatory and apoptotic mechanisms of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity in a model of Parkinson's disease. Our results showed that mutant mice lacking the caspase-11 gene were significantly more resistant to the effects of acute treatment with MPTP than their wild-type mice. Thus, the neurotoxicity of MPTP seems to be mediated by the induction of both mitochondrial dysfunction and free radical generation. Previously, we showed that overexpression of the Apaf-1 dominant-negative inhibitor inhibited the mitochondrial apoptotic cascade in chronic MPTP treatment but not in acute MPTP treatment. The present results indicate that MPTP neurotoxicity may be mediated via activation of the caspase-11 cascade and inflammatory cascade, as well as the mitochondrial apoptotic cascade.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

PubMed2003

Accurate estimation of forest carbon stocks by 3-D remote sensing of individual trees.

Forests are one of the most important carbon sinks on Earth. However, owing to the complex structure, variable geography, and large area of forests, accurate estimation of forest carbon stocks is still a challenge for both site surveying and remote sensing. For these reasons, the Kyoto Protocol requires the establishment of methodologies for estimating the carbon stocks of forests (Kyoto Protocol, Article 5). A possible solution to this challenge is to remotely measure the carbon stocks of every tree in an entire forest. Here, we present a methodology for estimating carbon stocks of a Japanese cedar forest by using a high-resolution, helicopter-borne 3-dimensional (3-D) scanning lidar system that measures the 3-D canopy structure of every tree in a forest. Results show that a digital image (10-cm mesh) of woody canopy can be acquired. The treetop can be detected automatically with a reasonable accuracy. The absolute error ranges for tree height measurements are within 42 cm. Allometric relationships of height to carbon stocks then permit estimation of total carbon storage by measurement of carbon stocks of every tree. Thus, we suggest that our methodology can be used to accurately estimate the carbon stocks of Japanese cedar forests at a stand scale. Periodic measurements will reveal changes in forest carbon stocks.

Automation↗

PubMed2002

IgG-immunostaining in the intact rabbit brain: variable but significant staining of hippocampal and cerebellar neurons with anti-IgG.

A significant number of brain neurons in the rabbit brain were immunostained with anti-rabbit gamma-immunoglobulin (IgG). IgG-positive neurons were often found in the cerebellum, lower brainstem and motor nuclei. Similar IgG-positive neurons were occasionally found in the hippocampus, cerebral cortex and midbrain, but not in the striatum and thalamus. These neurons showed very clear Golgi-like staining of soma and dendrites but IgG staining was absent from the cell nuclei and axons. In particular, groups of Purkinje neurons in the rabbit cerebellum showed strong IgG-positive staining. To confirm whether the staining reflected the existence of IgG molecules in these neurons, staining specificity was carefully evaluated. Staining was specifically eliminated by pre-absorption of the antibodies with the purified rabbit IgG. An antibody to the neural cell adhesion molecule (NCAM or CD56), a member of the immunoglobulin superfamily, exhibited a completely different pattern of staining as that for IgG. To determine whether IgG-like immunoreactivity was a general feature of mammalian brain, brain sections of rabbits, rats, and mice were immunostained with antibodies to IgGs of each of the three species. Similar IgG-positive neurons were observed in all three species, although the distribution and frequency was characteristic for each species. In rabbit brain, anti-rabbit IgG stained-neurons were more abundant compared to rat and mouse brain. IgG-positive microglia-like cells were evident in mouse brain, but less frequent in rabbit and were hardly observed in rat brain. To evaluate whether stained neurons could synthesize IgG, in situ hybridization was carried out using an antisense oligonucleotide probe to rabbit IgG DNA. No significant label was observed in cerebellum. These results suggest that a significant number of neurons in the intact rabbit brain take up IgGs and concentrate them in their cytoplasm, although the molecular uptake mechanism is retained for future studies. Our results also suggest that the rabbit may be a suitable animal to study the function(s) of IgG in brain neurons.

Animals↗

PubMed2002

Novel monoamine oxidase inhibitors, 3-(2-aminoethoxy)-1,2-benzisoxazole derivatives, and their differential reversibility.

Although possible usefulness of non-selective monoamine oxidase (MAO) inhibitors for Parkinson's disease therapy has been suggested in the literature, MAO inhibitors whose inhibition is reversible and have dual action to both MAO-A and -B subtypes is not available yet. Subtype selectivity and reversibility of a series of novel MAO inhibitors, 3-(2-aminoethoxy)-1,2-benzisoxazole derivatives, were studied. Several dual MAO inhibitors, which inhibit both MAO-A and -B, were obtained. When administered to mice, their effects were generally reversible. Among the derivatives, RS-1636 and RS-1653 had much longer duration of brain MAO-B inhibition than that of MAO-A. In vitro, the inhibited MAO-A activity by these compounds was partially recovered by buffer change at 4 degrees C, while little MAO-B activity was recovered. Although it is not fully elucidated yet, the reversibility of these inhibitors is probably determined primarily by this dissociation profile. This unique differential reversibility indicates that optimization of the balance of actions can be achieved by differentiating reversibility to each target molecule.

Animals↗