It has been shown that human monocytes express rnonocyte chemoattractant protein-1 (MCP-1), an inflammatry factor, in response to non-fibrillar P-amyloid protein. Reactive niicroglia and inflammatory factors were reported to be present in P-amyloid deposits (senile plaques) in Alzheimer's disease, suggesting the presence of MCP-1 in senile plaques. To address this issue, we examined MCP-1-immunoreactivity in senile plaques using a mouse monoclonal anti-MCP-1 antibody. Monocyte chemoattractant protein-1 was found immunohistochemically in mature senile plaques and reactive microglia but not in immature senile plaques of brain tissues from five patients with Alzheimer's disease. These findings suggest that MCP-1-related inflammatory events induced by reactive microglia contribute to the maturation of senile plaques.

It has been reported that many tau sites in neurofibrillary tangles (NFT) are abnormally phosphorylated. We investigated the phosphorylation of tau in the hippocampus of nondemented patients and Alzheimer''s disease patients by immunostaining with five site-specific antibodies against phosphorylated tau. In the pretangle stage, tau in neuropil threads was phosphorylated at serines 199, 202 and 409, numbered according to the longest human tau isoform, whereas tau in some neuronal soma was phosphorylated at serines 199, 202, 409 and 422. Tau at the stage of NFT was phosphorylated at serine 396 and threonine 231 in addition to serines 199, 202, 409 and 422. In the advanced stage, tau in ghost tangles was phosphorylated mainly at serine 396. These results suggest that the phosphorylation of each site in tau differs among the maturing stages of neurofibrillary change and that abnormal phosphorylation of tau in the neuronal soma occurs at 199, 202, 409 and 422 earlier than at threonine 231 and serine 396.

Our results suggest that HLA-B*59:01 is a risk factor for CIAG in the Japanese population. Furthermore, if our model is true, the results suggest that rechallenging certain CIG subjects with clozapine may not be always contraindicated.

Abstract:To examine the physiological roles of the ␦ subunit of Ca 2ϩ /calmodulin-dependent protein kinase II (CaM kinase II␦) in brain, we examined the localization of CaM kinase II␦ in the rat brain. A specific antibody to CaM kinase II␦1-␦4 isoforms was prepared by immunizing rabbits with a synthesized peptide corresponding to the unique carboxyl-terminal end of these isoforms. The prepared antibody did not recognize the ␣, ␤, and ␥ subunits, which were each overexpressed in NG108-15 cells. Immunoblot analysis on various regions and the nuclear fractions from rat brains suggested that some isoforms of CaM kinase II␦1-␦4 were abundant in the nucleus in the cerebellum. Total RNA from the cerebellum was analyzed by RT-PCR with a primer pair from variable domain 1 to variable domain 2. We detected the three PCR products ␦3.1, ␦3.4, and ␦3 that contained the nuclear localization signal. These CaM kinase II␦3 isoforms were localized in the nuclei in transfected NG108-15 cells. Immunohistochemical study suggested the existence of these isoforms in the nuclei in cerebellar granule cells. These results suggest that CaM kinase II␦3 isoforms are involved in nuclear Ca 2ϩ signaling in cerebellar granule cells. Key Words: Calmodulin-Ca 2ϩ /calmodulindependent protein kinase II-Cerebellum-Granule cells-Nucleus-Overexpression. J. Neurochem. 72, 815-825 (1999). Ca2ϩ /calmodulin-dependent protein kinase II (CaM kinase II) is highly concentrated in neuronal tissues and may be related to many neuronal functions (for review, see Colbran et al., 1989;Hanson and Schulman, 1992;Fukunaga et al., 1996). Four subunits, termed ␣, ␤, ␥, and ␦, are encoded by distinct genes in eukaryotes Schulman and Hanson, 1993). The ␥ and ␦ subunits appear to have widespread distribution, whereas the ␣ and ␤ subunits are predominant forms in the brain. All subunits are highly conserved and have Ͼ90% amino acid sequence homology in the Nterminal catalytic domain and in the regulatory domain comprising calmodulin-binding and autoinhibitory sequences. The various isoforms differ mainly at the end of the regulatory domain, an area with a variable number of amino acids (variable domain 1). The important findings were that 11-amino acid insertion in variable domain 1 functioned as a nuclear localization signal (NLS) (Srinivasan et al., 1994). Among the subunits of CaM kinase II found in the brain, only the ␣B subunit (corresponding to the ␣-33 subunit in monkey brain) and the ␥A subunit in rat brain had the NLS (Tobimatsu et al., 1988;Benson et al., 1991;Brocke et al., 1995).Northern blot analysis indicated that a significantly higher amount of the ␦ subunit of CaM kinase II (CaM kinase II␦) was expressed in cerebellum than in cerebrum and brainstem (Tobimatsu and Fujisawa, 1989). In situ hybridization histochemistry revealed intense expression signals for CaM kinase II␦ in the cerebellar granule cell layer, but not in the Purkinje cell layer (Takaishi et al., 1992;Sakagami and Kondo, 1993). These results suggest that CaM kinase II␦ was mainly expressed in ...

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