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We evaluated the immunohistochemical intensities of α-synuclein, phosphorylated α-synuclein (p-syn), dopamine- and cAMP-regulated phosphoprotein of 32 kDa (DARPP-32), calbindin-D 28k, calpain-cleaved carboxy-terminal 150-kDa spectrin fragment, and tyrosine hydroxylase in multiple system atrophy (MSA). The caudate head, anterior putamen, posterior putamen, substantia nigra, pontine nucleus, and cerebellar cortex from six MSA brains, six age-matched disease control brains (amyotrophic lateral sclerosis), and five control brains were processed for immunostaining by standard methods. Immunostaining for α-synuclein, p-syn, or both was increased in all areas examined in oligodendrocytes in MSA. Immunostaining for DARPP-32 and calbindin-D 28k was most prominently decreased in the posterior putamen, where neuronal loss was most prominent. Immunostaining for DARPP-32 and calbindin-D 28k was also diminished in the anterior putamen and caudate head, where neuronal loss was less prominent or absent. Calbindin immunostaining was also decreased in the dorsal tier of the substantia nigra and cerebellar cortex. Loss of immunostaining for DARPP-32 and calbindin-D 28k compared with that of neurons indicates calcium toxicity and disturbance of the phosphorylated state of proteins as relatively early events in the pathogenesis of MSA.

Most of the patients with Parkinson’s disease (PD) are sporadic. However, Since identification of monogenic forms of PD, the contribution of genetic factors to the pathogenesis of sporadic PD is proposed as one of major risk factors. Indeed, this is supported by the demonstration of the high concordance in twins, increased risk among relatives of PD patients in case control and family studies. Thus, the functional analysis for the gene products for familial PD provides us a good hint to elucidate the pathogenesis of nigral degeneration. For example, although α-synuclein is involved in a rare dominant form of familial PD with dopa responsive parkinsonian features, this molecule is a major component of and Lewy bodies (LBs). In contrast, Park2 (parkin-related disease) is the most frequent form among patients with young-onset PD. However, Park2 brains generally lack the formation of LBs. In the other word, parkin responsible for Park2 is essential for the formation of LBs. Thus, both α-synuclein and parkin are speculated to share a common pathway. Here, we reviewed the parkin function and molecular mechanisms of Park2.

To date 11 forms of familial Parkinson's disease (PD) have been mapped to different chromosome loci, of which 6 genes have been identified as the causative genes, i.e., alpha-synuclein (SNCA), parkin, UCH-L1, PINK1, DJ-1, and LRRK2. For UCH-L1, additional families with this mutation are necessary before concluding that UCH-L1 is the definite causative gene for PARK5, as only one family so far has been reported. SNCA, UCH-L1, and LRRK2 mutations cause autosomal dominant PD and the remaining gene mutations autosomal recessive PD. Age of onset tends to be younger in familial PD compared with sporadic PD, particularly so in autosomal recessive PD. Generally familial cases respond to levodopa quite nicely and progression of the disease tends to be slower. It is an interesting question how familial PD-causing proteins are mutually related each other. In this article, we review recent progress in genetics and molecular biology of familial PD.

We review recent progress in gene therapy utilizing experimental parkinsonian models including our data. Investigation of ex vivo gene therapy for Parkinson’s disease (PD) is to provide L-dopa by transplantation of genetically modified cells into the striatum. Recently, neuronal progenitor cells (NPC) are recognized as the most appropriate target population for such genetic and cellular therapy of PD. We have developed modified pseudo-typed retrovirus production system. Using this gene transfer system, it is easy and efficient to introduce the gene into NPC because high titer virus vector is easily obtained. For the in vivo gene therapy, adeno-associated virus (AAV) vector is best virus vector because it is easy to introduce gene into neurons without inflammatory reaction. We established in vivo models of the inhibition of the caspase-cascade by overexpression of apoptotic protease activating factor-1-dominant negative inhibitor (Apaf-1-DN) using AAV vector. We showed that Apaf-1-DN delivery using an AAV vector system could prevent nigrostriatal degeneration in MPTP mice, suggesting that it might be an antimitochondrial apoptotic gene therapy for PD.

Parkinson’s disease is a common neurodegenerative disease with complex clinical features. Recently, we idenfied a novel gene named Parkin to be responsible for the pathogenesis of autosomal recessive juvenile parkinsonism (AR-JP). Various mutations were found in AR-JP patients of Japanese and other ethnic origins, providing a definitive evidence for the Parkin to be a causative gene for AR-JP. The predicted structure of Parkin protein and its mutation provide important clues for studying the functional role of the Parkin protein in leading to selective degeneration of nigral neurons in the brains of AR-JP patients.

We review here familial Parkinson’s disease (PD) from clinical as well as molecular genetic aspects. The contribution of genetic factors to the pathogenesis of PD is supported by the demonstration of the high concordance in twins, increased risk among relatives of PD patients in case control and family studies, and the existence of familial PD and parkinsonism based on single gene defects. Recently, several genes have been mapped and/or identified in patients with familial PD. a-Synuclein is involved in a rare dominant form of familial PD with dopa responsive parkinsonian features and Lewy body positive pathology. In contrast, parkin is responsible for autosomal recessive form of early-onset PD with Lewy body-negative pathology. This form is identified world-wide among patients with young-onset PD. Furthermore, ubiquitin carboxy terminal hydrolase Ll gene is responsible for an autosomal dominant form of typical PD, although only a single family has so far been identified with a mutation of this gene, and tau has been identified as a causative gene for frontotemporal dementia and parkinsonism. In addition, five other chromosome loci have been identified to be linked to familial PD or dystonia-parkinsonism. The presence of different loci or different causative genes indicates that PD is not a single entity but a highly heterogeneous. Identification and elucidation of the causative genes should enhance our understanding of the pathogenesis of sporadic PD.

Although the exact mechanism of nigral cell death in Parkinson’s disease (PD) is not known, increasing evidence suggests the presence of apoptotic cell death in PD. When we applied the TUNEL method to detect DNA fragmentation, four out of seven late onset sporadic patients with PD showed TUNEL-positive neurons. The percentages of those neurons among the remaining melanin containing neurons were 0.6 to 4.8% (average 2.1%). But TUNEL-positive neurons could not be detected in control subjects as well as four patients with young onset (under 40 years of the age) PD. Numbers of nigral toxins such as MPTP, complex I inhibitors, and mitochondrial respiratory inhibitors have been reported to induced apoptotic cell death. These findings suggest that apoptosis is involved in nigral cell cleath in PD at least in part and warrant further studies on apoptosis-related substances in PD.

Numbers of biochemical abnormalities which may be relevant to the degenerative process of nigral dopaminergic neurons have been described. These include accumulation of iron in substantia nigra, decrease in the enzymatic activity and the amount of subunits of mitochondrial complex I, increase in the amount of deleted mitochondrial DNA, and possible increase in oxygen derived free radicals. Recent progress in this field is reviewed in this communication. Although the primary cause of Parkinson’s disease is still unknown, these abnormalities listed above will contribute to the progression of the degenerative process. In addition, we report our recent data on the toxic effects of iron and synthetic dopamine melanin on cultured dopaminergic neurons, and discuss possible interaction of iron and mitochondrial DNA.

Studies on mitochondrial respiratory enzymes in Parkinson’s disease are reviewed. The presence of abnormalities in Complex I of the mitochondrial respiratory chain was described in Parkinson’s disease by several groups of investigators utilizing different methods. These abnormalities do not seem to be a primary defect of Parkinson’s disease. Instead, they seem to be a consequence of long-standing noxious stimuli to substantia nigra. Dopaminergic neurons in substantia nigra appear to have a unique property with reduced tolerance to various noxious stimuli. Further studies on mitochondria seem to be important for the elucidation of Parkinson’s disease.