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Gene expression of soluble guanylate cyclase in the brain of adult rats
s13s EXPRESSION OF HIGHLY POLYSIALYLATED NCAM-120 IN A CALCITONIN-PRODUCING CELL LINE. ICHIRO NISHIYAMA 1, TATSUNORI SEKIZ. TADACHIKA OOTAJ, MASAKO OHTA4 AND MANABU OGISO4. 1Dept. of Pharmacol., Teikyo Univ. Sch. of Med.. Tokyo 173, ZDept.. of Anat.. Juntendo Univ. Sch. of Med.. Tokyo 113. JIsotope Cent.., Tokyo Univ. of Agri., Tokyo 156, 4Dept. of Phgsiol., Toho Univ. Sch. of Med., Tokyo 143. Japan
5-25
The thyroid C-cell (calcitonin-producing cell) is an endocrine cell derived from the neural crest,. The expression of the neural cell adhesion molecule (NCAM) was examined in rMTC 6-23 cells, Two monoclonal antibodies (MAbs) which a C-cell line derived from rat medullary thyroid carcinoma. recognize different epitopes in NCAM were used for the experiments: MAb 12E3 specifically reacts with the polysialic acid portion of highly polysialylated rat. NCAM (NCAM-H), and the other MAb recognizes polypeptide portion of all the three isoforms of rat. NCAM. About a half of rMTC 6-23 cells were found to be NCAM-H positive on their surfaces by immunocytochemistry. The molecular form of NCAM was analyzed by Western blotting. Immunoreactive NCAM was detected as a broad band around 200-250K using either of the MAbs. When the NCAM preparation was pretreated with neuraminidase, NCAM-H immunoreactivity was completely abolished, and concomitantly immunoreactive peptide which was of similar size to neuronal NCAM-120 was detected on the blot by MAb against NCAM peptides. These results indicate rMTC 6-23 cells express on their surfaces predominantly NCAM-120 which was highly polysialylated.
S-26
GENE EXPRESSION OF SOLUBLEGLIANYLATE CYCLASE IN THE BRAIN OF ADULT RATS KUNIHIKOUMEZAWA,KAORUGOTO, HIROYUKI SAKAGAMI, and mTAKE
KONDO Department of Anatomy, Tohoku University School of Medicine, 2-l Seiryo-cho, Aobamku, Sendai 981, JAPAN triggered by NO and cyclic GMP In order to understand how cGMP synthesis in the brain, the detailed localization of gene expression for soluble function guanyiate cyclase (sGC) was studied by in situ hybridization histochemistry using a 3’ noncoding region of PCR-amplified cDNA encoding 70K sGC. Intense hybridization signals were detected in the hippocampal pyramidal and dentate granule cell layers, and the cerebellar Purkinje cells. Moderate hybridization signals were found in the olfactory mitral cell layer, caudate putamen, substantia nigra, pars compacta, and medial septal nucleus.Weak to moderate gene expression for this enzyme was detected !n the piriform cortex,neocortex, superior and inferior colliculi,deep cerebellar and some brain stem nuclei such as pontine nucleus and occulomoter nuclei, nucleus. Major white matters such as corpus callosum and anterior this comissure expressed gene faintly.
5-27 Localization of cardiac muscle/skeletal muscle ryanodine receptor and cardiac muscle ryanodine receptor in mouse central nervous system.SETSUKONAKANISH~.GORO KUWAJIMA * AND KATSUHIKO MIKOSHIBA’, ~Pharmaceutical Basic Research Laboratories. JT INC.. Kanazawa-ku. Yokohama, 236. ZShionogi Institute for Medical Science, Mishima, Settu. Osaka 566. jInstitute of Medical Science, Universitv of Tokvo, Minato-ku, Tokyo, 108. Ryanodine receptor-like immunoreactivity in the mouse central nervous system was presented using two antibodies, C2 and C4, which were raised against two synthetic peptides. C2 recognized cardiac muscle and skeletal muscle ryanodine receptor. C4 recognized cardiac muscle ryanodine receptor. Western blotting analysis and [3H]ryanodine binding assay showed high content of cardiac muscle ryanodine receptor in hippocampus and cerebral cortex but was lower in other regions of the brain. Immunohistochemical study demonstrated that a wide spread distribution of these receptors in various regions of the brain and spinal cord. In hippocampus, pyramidal neurons in CA2-3 region were more labeled than CA1 region. In cerebellum, the Purkinje cells, the Golgi cells, the granule cells as well as the neurons of cerebellar nuclei were labeled. Immunoreactive sites were unevenly distributied in somata. The fibers were more immunoreactive with the C4 antibody than the C2 antibody. The localization of ryanodine receptors differed from that of inositol I ,4.5-trisphosphate receptros in the central nervous system.
5-25
The thyroid C-cell (calcitonin-producing cell) is an endocrine cell derived from the neural crest,. The expression of the neural cell adhesion molecule (NCAM) was examined in rMTC 6-23 cells, Two monoclonal antibodies (MAbs) which a C-cell line derived from rat medullary thyroid carcinoma. recognize different epitopes in NCAM were used for the experiments: MAb 12E3 specifically reacts with the polysialic acid portion of highly polysialylated rat. NCAM (NCAM-H), and the other MAb recognizes polypeptide portion of all the three isoforms of rat. NCAM. About a half of rMTC 6-23 cells were found to be NCAM-H positive on their surfaces by immunocytochemistry. The molecular form of NCAM was analyzed by Western blotting. Immunoreactive NCAM was detected as a broad band around 200-250K using either of the MAbs. When the NCAM preparation was pretreated with neuraminidase, NCAM-H immunoreactivity was completely abolished, and concomitantly immunoreactive peptide which was of similar size to neuronal NCAM-120 was detected on the blot by MAb against NCAM peptides. These results indicate rMTC 6-23 cells express on their surfaces predominantly NCAM-120 which was highly polysialylated.
S-26
GENE EXPRESSION OF SOLUBLEGLIANYLATE CYCLASE IN THE BRAIN OF ADULT RATS KUNIHIKOUMEZAWA,KAORUGOTO, HIROYUKI SAKAGAMI, and mTAKE
KONDO Department of Anatomy, Tohoku University School of Medicine, 2-l Seiryo-cho, Aobamku, Sendai 981, JAPAN triggered by NO and cyclic GMP In order to understand how cGMP synthesis in the brain, the detailed localization of gene expression for soluble function guanyiate cyclase (sGC) was studied by in situ hybridization histochemistry using a 3’ noncoding region of PCR-amplified cDNA encoding 70K sGC. Intense hybridization signals were detected in the hippocampal pyramidal and dentate granule cell layers, and the cerebellar Purkinje cells. Moderate hybridization signals were found in the olfactory mitral cell layer, caudate putamen, substantia nigra, pars compacta, and medial septal nucleus.Weak to moderate gene expression for this enzyme was detected !n the piriform cortex,neocortex, superior and inferior colliculi,deep cerebellar and some brain stem nuclei such as pontine nucleus and occulomoter nuclei, nucleus. Major white matters such as corpus callosum and anterior this comissure expressed gene faintly.
5-27 Localization of cardiac muscle/skeletal muscle ryanodine receptor and cardiac muscle ryanodine receptor in mouse central nervous system.SETSUKONAKANISH~.GORO KUWAJIMA * AND KATSUHIKO MIKOSHIBA’, ~Pharmaceutical Basic Research Laboratories. JT INC.. Kanazawa-ku. Yokohama, 236. ZShionogi Institute for Medical Science, Mishima, Settu. Osaka 566. jInstitute of Medical Science, Universitv of Tokvo, Minato-ku, Tokyo, 108. Ryanodine receptor-like immunoreactivity in the mouse central nervous system was presented using two antibodies, C2 and C4, which were raised against two synthetic peptides. C2 recognized cardiac muscle and skeletal muscle ryanodine receptor. C4 recognized cardiac muscle ryanodine receptor. Western blotting analysis and [3H]ryanodine binding assay showed high content of cardiac muscle ryanodine receptor in hippocampus and cerebral cortex but was lower in other regions of the brain. Immunohistochemical study demonstrated that a wide spread distribution of these receptors in various regions of the brain and spinal cord. In hippocampus, pyramidal neurons in CA2-3 region were more labeled than CA1 region. In cerebellum, the Purkinje cells, the Golgi cells, the granule cells as well as the neurons of cerebellar nuclei were labeled. Immunoreactive sites were unevenly distributied in somata. The fibers were more immunoreactive with the C4 antibody than the C2 antibody. The localization of ryanodine receptors differed from that of inositol I ,4.5-trisphosphate receptros in the central nervous system.