- Email: [email protected]
814 Effect of chronic morphine administration on gene expressions in mouse amygdala complex
s94 EXPRESSION OF CALCIUM-RESPONSIVE GENES WITH AN ACTIVIN-DEPENDENT SURVIVAL OF CULTURED CEREBELLAR GRANULE CELLS. MASAAKI TSUDA, AKIKO TABUCHI. KUNIAKI SAN0 Fat. of Pharmaceu. Sci., Okayama Univ. Okayama 700, Japan 812
Rescue of cerebellar granule cells from the cell death process induced by the exposure of cells to 5 mM KCI for 24 hrs in culture was attained by a secondary stimulation of cells with 25 mM KCI, in concert with a dramatic increase in brain-derived neurotrophic factor (BDNF) mRNA expression which was dependent upon Caz+ influx into the cells. However, the addition of BDNF only partly rescued the cells. The cells cultured with 25 mM KCI were more resistent to glutamate excitotoxicity than those cultured with 5 mM KCI. The expression of neurotrophin-3 (NT-3) gene was found to be suppressed under the culture condition including 25 mM KCI. We are now searching for calcium-responsive genes whose expression could concurrently be changed by the intracellular Caz+ signals.
813
CLONING OF cDNA FOR A RETINA-SPECIFIC SERINE PROTEASE AND ITS mRNA EXPRESSION NNABU TANIGUCHI, SHIGETAKA YOSHIDA, SADAO SHIOSAKA, Department of Structural Cellular Biology, NAIST, Ikoma, Nara, 630.01, Japan To clone a retina-specific serine protease, we screened the cDNA library from the mouse eyes using a RT-PCR-amplified DNA fragment as a probe. Sequence analysis of the positive clones and search in the database showed that the clones are cDNAs for the mouse homolog of RNK-Met-l. RNK-Met-I is a serine protease rat leukemia natural killer cells. In Northern blotting and RT-PCR analysis the mouse Met-ase(MR-Met-l) mRNA expression was detected exclusively in the eye. In situ hybridization histochenistry showd that the mRNA expressed strongry in the pigment epithelial layer, photoreceptor inner segment. Hybridization layer. In the ontogenetical study, the expression and increased during maturation.(the significance
signals were also detected in innner granular of the mRNA was first detected at postnatal day 5 of the expression of serine protease will
bediscussed.)
814
OF CHRONIC MORPHINE ADMINISTRATION EFFECT MOUSE AMYGDALA COMPLEX. IKEMOTO MITSUSHI.
Dent. of Biomolecular Technoloav, AIST, MITI.
Enaineerina, National Institute Tsukuba, Ibaraki, 305, Japan.
ON
of
GENE
EXPRESSIONS
Bioscience
IN
and Human-
It has been speculated that the regulation of gene expression may be involved in We have recently demonstrated that the chronic morphine tolerance and dependence. changes gene expressions in mouse amygdala complex (1). To morphine administration to construct the subtractive clarify the chronic morphine-induced genes, I tried cDNA library from the mice amygdala complex mRNAs extracted at 16 hours after the differential hybridization, I finally isolated 11 last administration. By using In situ hybridization analysis revealed that the increase of positive cDNA clones. 01 mRNA was detected at the basolateral amygdala nucleus only by chronic morphine treatment. It was completely blocked by co-administration of naloxone. Furthermore, dentate such as cerebral cortex, 01 mPNA was localized in the brain regions cerebellum and locus coelurus. Northern blot and RT-PCR hippocampus (CAl-3), gyms, 01 mRNA might be localized in glial cells. These results analysis suggested that in suggest that chronic morphine administration elicits a change of gene expressions I am trying to analyze this clone in detail. glial cells of mouse amygdala complex. (1) Ikemoto,M. et al. NeuroReport, 6, 262-264,(1995).
Rescue of cerebellar granule cells from the cell death process induced by the exposure of cells to 5 mM KCI for 24 hrs in culture was attained by a secondary stimulation of cells with 25 mM KCI, in concert with a dramatic increase in brain-derived neurotrophic factor (BDNF) mRNA expression which was dependent upon Caz+ influx into the cells. However, the addition of BDNF only partly rescued the cells. The cells cultured with 25 mM KCI were more resistent to glutamate excitotoxicity than those cultured with 5 mM KCI. The expression of neurotrophin-3 (NT-3) gene was found to be suppressed under the culture condition including 25 mM KCI. We are now searching for calcium-responsive genes whose expression could concurrently be changed by the intracellular Caz+ signals.
813
CLONING OF cDNA FOR A RETINA-SPECIFIC SERINE PROTEASE AND ITS mRNA EXPRESSION NNABU TANIGUCHI, SHIGETAKA YOSHIDA, SADAO SHIOSAKA, Department of Structural Cellular Biology, NAIST, Ikoma, Nara, 630.01, Japan To clone a retina-specific serine protease, we screened the cDNA library from the mouse eyes using a RT-PCR-amplified DNA fragment as a probe. Sequence analysis of the positive clones and search in the database showed that the clones are cDNAs for the mouse homolog of RNK-Met-l. RNK-Met-I is a serine protease rat leukemia natural killer cells. In Northern blotting and RT-PCR analysis the mouse Met-ase(MR-Met-l) mRNA expression was detected exclusively in the eye. In situ hybridization histochenistry showd that the mRNA expressed strongry in the pigment epithelial layer, photoreceptor inner segment. Hybridization layer. In the ontogenetical study, the expression and increased during maturation.(the significance
signals were also detected in innner granular of the mRNA was first detected at postnatal day 5 of the expression of serine protease will
bediscussed.)
814
OF CHRONIC MORPHINE ADMINISTRATION EFFECT MOUSE AMYGDALA COMPLEX. IKEMOTO MITSUSHI.
Dent. of Biomolecular Technoloav, AIST, MITI.
Enaineerina, National Institute Tsukuba, Ibaraki, 305, Japan.
ON
of
GENE
EXPRESSIONS
Bioscience
IN
and Human-
It has been speculated that the regulation of gene expression may be involved in We have recently demonstrated that the chronic morphine tolerance and dependence. changes gene expressions in mouse amygdala complex (1). To morphine administration to construct the subtractive clarify the chronic morphine-induced genes, I tried cDNA library from the mice amygdala complex mRNAs extracted at 16 hours after the differential hybridization, I finally isolated 11 last administration. By using In situ hybridization analysis revealed that the increase of positive cDNA clones. 01 mRNA was detected at the basolateral amygdala nucleus only by chronic morphine treatment. It was completely blocked by co-administration of naloxone. Furthermore, dentate such as cerebral cortex, 01 mPNA was localized in the brain regions cerebellum and locus coelurus. Northern blot and RT-PCR hippocampus (CAl-3), gyms, 01 mRNA might be localized in glial cells. These results analysis suggested that in suggest that chronic morphine administration elicits a change of gene expressions I am trying to analyze this clone in detail. glial cells of mouse amygdala complex. (1) Ikemoto,M. et al. NeuroReport, 6, 262-264,(1995).