Flexible fate commitment of E2-2high common DC progenitors implies tuning in tissue microenvironments.

The basic helix-loop-helix transcription factor E2-2 is essential for the development of plasmacytoid dendritic cells (pDCs) but not conventional DCs (cDCs). Here, we generated E2-2 reporter mice and demonstrated that an E2-2high fraction among common DC progenitors, which are a major source of pDCs and cDCs in the steady state, strictly gave rise to pDCs in the presence of Flt3 (Fms-like tyrosine kinase receptor-3) ligand ex vivo or in the secondary lymphoid organs when transferred in vivo. However, in the small intestine, some of these E2-2high progenitors differentiated into cDCs that produced retinoic acid. This transdifferentiation was driven by signaling via the common ß receptor, a receptor for the cytokines IL-3, IL-5 and GM-CSF, which are abundant in the gut. In the presence of GM-CSF and Flt3 ligand, E2-2high-progenitor-derived cDCs consistently induced Foxp3+ Treg cells ex vivo. Our findings reveal the commitment and flexibility of E2-2high progenitor differentiation and imply that pertinent tuning machinery is present in the gut microenvironment.

Effect of angiotensin-converting enzyme inhibitor perindopril on interneurons in MPTP-treated mice.

We examined the effects of perindopril on the dopaminergic system in mice after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment. The mice received four intraperitoneal injections of MPTP at 1-h intervals. Administration of perindopril showed dose-dependent neuroprotective effects against striatal dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) depletion 3 days after MPTP treatment. Our immunohistochemical study showed that MPTP can severe damage in tyrosine hydroxylase (TH)-immunoreactive neurons after MPTP treatment. The administration of perindopril significantly attenuated MPTP-induced substantia nigra and striatal damage. The present study also showed that the immunoreactivity of parvalbumin (PV)- or neuronal nitric oxide synthase (nNOS)-positive cells in the substantia nigra was decreased 7 days after MPTP treatment, whereas no significant changes were observed in these cells of the striatum throughout the experiments. The administration of perindopril significantly attenuated MPTP-induced decrease of the PV- or nNOS-immunoreactivity in the nigral cells. In double-labeled immunostaining with anti-PV and anti-nNOS antibody, PV-immunoreactive cell bodies and fibers were not double-labeled for nNOS-immunoreactive cell bodies and fibers in both the striatum and substantia nigra after MPTP treatment. Furthermore, PV- or nNOS-immunoreactive cell bodies and fibers in both the striatum and substantia nigra were not double-labeled for TH-immunoreactive cell bodies and fibers. These results demonstrate that the ACE inhibitor perindopril has a dose-dependent protective effect against MPTP-induced striatal dopamine, DOPAC and HVA depletion in mice. The present study also demonstrates that perindopril is effective against MPTP-induced degeneration of the nigral neurons and interneurons. Furthermore, our immunohistochemical study suggests that PV-immunoreactive cells and nNOS-immunoreactive cells are different interneurons in both the striatum and substantia nigra. Thus, our results provide further evidence that the ACE inhibitor perindopril may offer a novel therapeutic strategy for Parkinson's disease (PD).

Arundic acid, an astrocyte-modulating agent, protects dopaminergic neurons against MPTP neurotoxicity in mice.

We examined the neuroprotective effects of a novel astrocyte-modulating agent, arundic acid (ONO-2506), in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of Parkinson's disease. Male C57BL/6 mice received four intraperitoneal injections of MPTP (20 mg/kg) at 2 h intervals. Dopamine content in the striatum was reduced to 21% of the normal control after 7 days. Treatment with arundic acid (30 mg/kg, i.p.) administered 1 min, 6 h, 24 h, 48 h, and 72 h after the last MPTP injection prevented the dopamine depletion (52% of the control, p<0.01). In addition, this treatment resulted in behavioral benefits. Behavioral testing showed that MPTP-injected mice exhibited motor deficits in the pole test and catalepsy test after 7 days, but arundic acid prevented the appearance of motor abnormalities in these tests. The MPTP-injected animals exhibited an 87% loss of tyrosine hydroxylase-containing dopaminergic neurons in the substantia nigra after 7 days, but the arundic acid-treated mice showed only a 56% reduction (p<0.01). GFAP-positive reactive astrocytes were accumulated in the striatum and substantia nigra 7 days after the MPTP injection, whereas arundic acid treatment induced an earlier appearance of reactive astrocytes by 3 days. The reactive astrocytes increased the production of S-100 protein, which is thought to promote neuronal damage, but arundic acid suppressed the expression of S-100. Thus, arundic acid protected dopaminergic neurons against MPTP neurotoxicity in mice and ameliorated neurological deficits. The results suggest that the neuroprotection is mediated through the modulation of astrocytic activation, including the inhibition of S-100 protein synthesis.

Biochemical, behavioral and immunohistochemical alterations in MPTP-treated mouse model of Parkinson's disease.

The biochemical, behavioral and immunohistochemical manifestations were investigated in mice subjected to four experimental schedules with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) hydrochloride treatment. The mice were treated intraperitoneally with MPTP (20 mg/kg in saline) four times a day at 2-h intervals showed severe and persistent depletions of dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the striatum and behavioral deficits, as compared with those (1) treated with MPTP (15 mg/kg in saline ip) once a day for 14 consecutive days; (2) MPTP (30 mg/kg in saline ip) twice a day for five consecutive days; and (3) MPTP (10 mg/kg in saline ip) four times a day at 1-h intervals for two consecutive days. The immunohistochemical study has shown that the acute treatment with MPTP caused severe loss of tyrosine hydroxylase (TH)- and dopamine transporter (DAT)-immunoreactive dopaminergic neurons and marked increase in glial fibrillary acidic protein (GFAP)-immunoreactive astrocytes in the striatum and the substantia nigra. Thus acute treatment of mice with MPTP was accompanied by sustained nigral degeneration and motor abnormalities. Furthermore, our results with Cu/Zn-superoxide dismutase (Cu/Zn-SOD) and manganese superoxide dismutase (Mn-SOD) immunostainings suggest that altered capacity of free radicals quenching may play a key role in the development of the neurons and interneuron damage after MPTP neurotoxicity. Thus, our findings provide valuable information on age-related disease progression and mechanisms of neurodegeneration.

Protective effects of neuronal nitric oxide synthase inhibitor in mouse brain against MPTP neurotoxicity: an immunohistological study.

We recently reported that neuronal nitric oxide synthase (nNOS) inhibitor, 7-nitroindazole, can protect against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) neurotoxicity in mice. It protected against both dopamine depletions and tyrosine hydroxylase (TH) positive neuron decreases in the mouse brain. In the present study, we further examined whether 7-nitroindazole can also protect against the alterations of TH-, microtubule-associated protein 2a,b (MAP2)-, glial fibrillary acidic protein (GFAP)-, parvalbumin (PV)-, dopamine transporter (DAT)-, nNOS- or endothelial NOS (eNOS)-positive cells, in comparison with pargyline as a relatively selective inhibitor of the monoamine oxidase-B (MAO-B). The present study showed that nNOS inhibitor as well as MAO-B inhibitor has a dose-dependent protective effect against MPTP-induced striatal dopamine and DOPAC depletion in mice. Furthermore, the present study revealed that 7-nitroindazole and pargyline can protect the alterations of immunohistochemical changes in the striatum and substantia nigra after MPTP treatment. These protective effects may be, at least in part, produced by the reduction of neuronally derived NO and peroxynitrite caused by MPTP. Our results also demonstrate that MPTP can cause functional damage of interneurons in the substantia nigra. These results suggest the possibility that nNOS inhibitors as well as MAO-B inhibitors may be therapeutically useful in neurodegenerative diseases such as Parkinson's disease. Thus our present results provide valuable information for the pathogenesis of degeneration of the nigrostriatal dopaminergic neuronal pathway.

Expression of S-100 protein is related to neuronal damage in MPTP-treated mice.

S-100beta is a calcium-binding protein expressed at high levels in brain and is known as a marker of brain damage. However, little is known about the role of S-100beta protein during neuronal damage caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). To determine whether S-100beta protein is induced in glial cells after MPTP treatment, we investigated the expression of S-100 protein immunohistochemically, using MPTP-treated mice. We also examined the change of neurons and glial cells in mice after MPTP treatment. The present study shows that tyrosine hydroxylase (TH) immunoreactivity decreased gradually in the striatum and substantia nigra from 1 day after MPTP treatment. Thereafter, TH-immunopositive cells and fibers decreased in the striatum and substantia nigra at 3 days after MPTP treatment. In contrast, S-100-immunopositive cells and glial fibrillary acidic protein (GFAP)-immunopositive cells increased markedly in the striatum and substantia nigra at 3 days after MPTP treatment. Seven days after MPTP treatment, S-100-immunopositive cells decreased in the striatum and substantia nigra. However, the number of GFAP-immunopositive cells increased in these regions. In double-labeled immunostaining with anti-S-100 and anti-GFAP antibodies, S-100 immunoreactivity was observed only in the GFAP-positive astrocytes. These results provide evidence that astrocytic activation may play a role in the pathogenesis of MPTP-induced degeneration of dopaminergic neurons. Furthermore, the present study demonstrates that S-100 protein is expressed selectively by astrocytes, but not by microglia, after MPTP treatment. These results provide valuable information for the pathogenesis of the acute stage of Parkinson's disease.

Effects of Ca2+ antagonists on motor activity and the dopaminergic system in aged mice.

We investigated the effects of the Ca(2+) antagonist nilvadipine on the dopaminergic system and motor activity in aged mice, in comparison with an other Ca(2+) antagonist, amlodipine. Furthermore, we examined the close correlation between the dopaminergic system and motor activity during the aging process. Striatal dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) contents were measured in 2-, 4-, 8-, 18- and 36-week-old mice. Behavioral tests (pole and catalepsy test) were performed with 4- and 36-week-old mice. Nilvadipine or amlodipine was administered intraperitoneally twice a day for 3 consecutive days to 30-36-week-old mice. The striatal dopamine, DOPAC and HVA contents were examined and behavioral tests were performed 1h after the last injection of each Ca(2+) antagonist. The dopamine, DOPAC and HVA contents in 2-week-old mice were significantly decreased in the striatum, as compared with 4-week-old animals. Thereafter, age-related increases in the dopamine, DOPAC and HVA contents were observed from 4 to 18 weeks old. However, in 36-week-old mice, the dopamine and DOPAC contents were reduced in the striatum, as compared with 18-week-old animals. Age-related decreases in motor function between 5- and 36-week-old mice were observed in both pole test and catalepsy tests. On the other hand, nilvalipine treatment produced a significant and dose-dependent increase in the striatal dopamine and DOPAC contents in 30-36-week-old mice. In contrast, no significant changes were observed in the striatal dopamine content in amlodipine-treated mice, although this drug showed a significant and dose-dependent increase in the striatal DOPAC and HVA content. In our behavioral study, nilvadipine also showed a significant and dose-dependent inhibition against motor deficits in 30-36-week-old mice. In contrast, amlodipine showed no significant effect on motor deficits in 30-36-week-old mice. The present study demonstrated that nilvadipine has a protective effect against the deficits in both the striatal dopaminergic system and motor activity in aged mice. Our study also suggested that the beneficial effect of nilvadipine against motor abnormalities may be mediated by a protective effect against the reduced activity of the dopaminergic system in aged mice. These results suggested that nilvadipine may offer a new approach for the treatment of hypobulia in aged humans.