Binding of clathrin assembly protein to clathrin triskelia induces their assembly into clathrin-coated vesicle in neurons. The clathrin binding protein is a neuronal-specific, synapse associated protein that is expressed nonuniformly in rat brain. We isolated two cDNAs, encoding the novel clathrin assembly protein, which has a 73% amino acid homology compared with that of AP180 protein when translated into amino acids. The deduced molecular weight is 64kD. The N-terminal domain harbouring clathrin binding site is very similar to that of AP180, and the C-terminal domain is much more different with that of AP180, which suggests that the novel protein mediates the assembly of clathrin and its regulatory role in the release of secretory vesicle.

Voltage-gated calcium channel (VGCC) is composed of at least four (α1, α2, β, and δ)subunits. Among them β subunit accelerates the kinetics of activation (channel opening) and nactivation (channel closure), and reguates the channel activity by phosphprylation through PKAand PKC that are activated by various signal transduction mechanisms. Until recently four isoforms of beta subunits (β1, β2, β3, β4) have been identified. Our recent data shows that VDCCβ3 gene is expressed only in e nervous system and around the perinatal stage at high level.Alternative splicing was o observed at both 5'- and 3'- ends. To elucidate alternative splicingand cts-acting element of gene regulation of the β3 subunit gene we isolated a 12.5kb-sizedgenomic clone encompassing β3 subunit gene from human genomic library using the whole β3subunit cDNA from NG108-l5 cell line as a probe. The genome was analyzed by Southem hybridization and sequencing. The β3 subunit gene consists at least of 12 exons, and deduced amino acid sequence from the exons showed 98% similarity with that of rat gene. The β3 subunitgene is not alternatively spliced at the middle of the gene, and has many possible phosphorylation sites,which may confer the regulatory role of the β3 subunit gene.

Cytochrome P-450(CYP) enzymes are important in catalyzing the hiotransffrmation on manyendogeneous compounds and xenobiotics, including drugs and carcinogens. In the presentstudy, effect of nifedipine a voltage dependent calcium channel blocker on the induction ofCYP1A1 and 2B1 was investigated. Change of CYP1A1 and 2B1 activities were measuredby using specific enzyme activities and Western blot analysis. CYP1A1, as quantified by ethoxyresorufin-0-deethylase activity and Western blot with monoclonal antibody 1-7-1, increasedin liver microsome of nifedipine-treated spontaneous hypertensive rat(SHR. 30mg/kg.b.w, twicea day for 3days) but not in kidney microsome. CYP2B1, as quantified by benzyloxyresorufin-O-dealkylase activity and Western blot wit]1 monoclonal antibody 2-66-3, markedly increasedin liver microsome of nifedipine-treated SHR but slightly in kidney microsome. The resultsdemonstrate that nifedipine is a potent inducer of CYP2B1 in SHR.

To develop the biomarkers for the immune dysfunction induced by dioxin, 2, 3, 7, 8-TCDD was administered C57BL/6N mice 0.1µg TCDD/??body weight. Colony forming assay showed that the proliferation potential of hemotopoieti progenitor cells in bone marrow was reduced 35-55% more by earlier exposure. The microarray experiments were duplicated, and the candidates were restricted for the genes expressed greater than 121-fold. Resulting candidates were 55 genes. The expression patterns of the whole genes were analyzed by self-organizing maps (SOM). From these results, we selected the stage-specific genes : one genes (SOM c21 : nadh dehydrogenase subunit 5 gene) for gestational 13.5day, 13 genes (c0 : sialophorin ; spn gene, etc) for postnatal 3 week. The above genes are proposed to be a potential use of biomarker for dioxin exposure in the case of immume dysfunctions.