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18. Proliferation and migration kinetics of stem cells in the rat fundic gland

Author

Yang, D.H., Tsuyama, S., Ge, Y. B., Wakamatsu, D., Ohmori, J., and Murata, F.

Subjects
Stem cell, 6 - Ciencias aplicadas::61 - Medicina [CDU], Cell kinetics
Abstract

The proliferation and migration of stem cells in the developing and adult rat fundic gland have been studied using BrdU immunohistochemistry and BrdUGSA 11 (Griffonia-simplicifolia agglutinin-11) double staining. In the developing rat fundic gland, stem cells were first scattered throughout al1 levels of the epithelia and then concentrated in the depth of the pits. With the elongation and maturation of the fundic glands, stem ceils left the gland base and moved upward. By 4 weeks after birth, the development of the fundic gland was completed and stem cells were confined to a narrow proliferative zone in the isthmus, reaching the adult distribution pattern. In the adult rat fundic gland, stem cells in the isthmus differentiated and migrated upward and downward, replacing the surface mucous cells and glandular cells respectively. For upward migration, it took about one week for stem cells to migrate from the isthmus to the surface. For downward migration, it took about two weeks for stem cells to migrate from the isthmus to the neck, and it took 30-36 weeks to reach the gland unit's blind end. FinaJly stem cells were lost at the deepest leve1 of the glands. The results obtained by simple topographical distribution in the present experiment agreed well with those obtained by quantitative analysis, suggesting the usefulness of BrdU immunohistochemistry for cell kinetic studies.

Published
1997

27. Abstracts

Author

Matsuno, Akira, primary, Nagashima, Tadashi, additional, Osamura, R.Yoshiyuki, additional, IKE, Hiromi, additional, TAMADA, Yoshitaka, additional, IIJIMA, Norio, additional, HAYASHI, Seiji, additional, TANAKA, Masaki, additional, ISHIHARA, Akihiko, additional, HASEGAWA, Michinori, additional, SUWA, Fumihiko, additional, IBATA, Yasuhiko, additional, IZUMI, Shin-ichi, additional, SENBA, Yuko, additional, SHIN, Masashi, additional, HISHIKAWA, Yoshitaka, additional, KOJI, Takehiko, additional, IWASAKA, Toshiki, additional, UMEMURA, Shinobu, additional, KAKIMOTO, Kochi, additional, TAKAHASHI, Akemi, additional, KOIZUMI, Haruko, additional, OSAMURA, R.Yoshiyuki, additional, Yamashita, Shuji, additional, Takayanagi, Atsushi, additional, Shimizu, Nobuyoshi, additional, ISHIZUYA-OKA, Atsuko, additional, UEDA, Shuichi, additional, Komiyama, Akira, additional, Katoh, Ryohei, additional, Yokoyama, Akira, additional, Kawaoi, Akira, additional, Torihashi, Shigeko, additional, Tsuyama, S., additional, Yang, D-H., additional, Murata, F., additional, TAJIMA, Hikaru, additional, ITOH, Masataka, additional, TAKAHASHI, Shinichi, additional, ISHIDA, Hitoshi, additional, SAITO, Syouzou, additional, KAWAKAMI, Hayato, additional, HIRANO, Hiroshi, additional, DAIMON, Tateo, additional, KAWAI, Kazuhiro, additional, KATAOKA, Katsuko, additional, SUZAKI, Etsuko, additional, Ito, Kinji, additional, Hoshida, Minako, additional, Hayashi, Michiko, additional, Ito, Athuko, additional, YANG, Dong-Hua, additional, Tsuyama, Shinichiro, additional, Murata, Fusayoshi, additional, MORIOKA, Kiyokazu, additional, Takano-Ohmuro, Hiromi, additional, ANDOH, Noriaki, additional, OHTANI, Haruo, additional, NAGURA, Hiroshi, additional, MIYOSHI, Norio, additional, KARAYA, Kazuhiro, additional, WATANABE, Masakatsu, additional, FUKUDA, Masaru, additional, Kobayashi, Toshihiro, additional, Okada, Teruhiko, additional, Seguchi, Harumichi, additional, Ueda, Tadashi, additional, Ishikawa, Yoshimaro, additional, Tsukinoki, Keiichi, additional, Imaizumi, Tomoko, additional, Miyoshi, Yoshiko, additional, Yamamoto, Takayuki, additional, Watanabe, Yoshihisa, additional, Karakida, Kazunari, additional, Iwasa, M, additional, Noriki, S, additional, Imamura, Y, additional, Fukuda, M, additional, KOMIYAMA, Kazuo, additional, Okaue, Masahiro, additional, Oda, Yasuyuki, additional, SATO, Junichi, additional, OKANO, Tadao, additional, MORO, Ram, additional, ITO, Atsuko, additional, HAYASHI, Michiko, additional, HOSHIDA, Minako, additional, ITO, Kinji, additional, CHIDA, Kohsuke, additional, MITSUMOTO, Yasuhide, additional, MORIGUCHI, Michihihsa, additional, NAKAJIMA, Tomoki, additional, HIKITA, Hiroshi, additional, OKANOUE, Takeshi, additional, ASHIHARA, Tsukasa, additional, SUGIHARA, Hiroyuki, additional, HATTORI, Takanori, additional, Hosokawa, Yohei, additional, Arai, Shouichi, additional, Ashihara, Tsukasa, additional, Tani, Naoki, additional, Taniguchi, Hiroki, additional, Nakanishi, Masayoshi, additional, Sakakura, Chouhei, additional, Mazaki, Takeshi, additional, Tsuchihashi, Yasunari, additional, Yamagishi, Hisakazu, additional, Nakamura, Nobuki, additional, Miyagi, Eri, additional, Suzuki, Koich, additional, Hemmi, Akihiro, additional, SAKAMOTO, Yoshika, additional, ITO, Takaaki, additional, OKUDELA, Koji, additional, KITAMURA, Hitoshi, additional, NAKANO, Kan-yu, additional, IYAMA, Ken-ichi, additional, Date, Fumiko, additional, Sasano, Hironobu, additional, Nagura, Hiroshi, additional, FURUTA, Hirokazu, additional, YOKOYAMA, Keiichi, additional, MASUDA, Shinsuke, additional, TAKAMATSU, Tetsuro, additional, TAJIMA, Y., additional, KAWASAKI, M., additional, OHNO, J., additional, KUSAMA, K., additional, MARUYAMA, S., additional, and UCHIDA, Kenzou, additional

Published
1999
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28. Abstracts

Author

Matsuo, Eiichi, primary, Furuno, Yukihiro, additional, Komatsu, Akio, additional, Maekawa, Suguru, additional, Murata, Kenichiroh, additional, Kikuchi, Taiyou, additional, SHIODA, Seiji, additional, NAKAI, Yasumitsu, additional, Yamashita, Shuji, additional, NAGATA, H., additional, TAKEKOSHI, S., additional, HASEGAWA, H., additional, ITOH, J., additional, YAMAMOTO, Y., additional, WATANABE, K., additional, FUSHIKI, Shinji, additional, KINOSHITA, Chikako, additional, NAGATA, Akihiro, additional, MAEDA, Toshihiro, additional, TOKUNAGA, Yoshimitsu, additional, MATSUMURA, Hitoshi, additional, KITAHAMA, Kunio, additional, SETO-OHSHIMA, Akiko, additional, KAWAMURA, Noriko, additional, TSUCHIHASHI, Yasunari, additional, MATSUMOTO, Takahiro, additional, MITSUFUJI, Shoji, additional, TOKITA, Kazuhiko, additional, MARUYAMA, Kyohei, additional, KODAMA, Tadashi, additional, ISEKI, Shoichi, additional, MABUCHI, Yoshio, additional, MARUYAMA, Hiromichi, additional, SAKUMA, Eisuke, additional, SOJI, Tsuyoshi, additional, OKADA, Teruhiko, additional, KOBAYASHI, Toshihiro, additional, ZINCHUK, Vadium S, additional, SEGUCHI, Harumichi, additional, DAIMON, Tateo, additional, OGUNI, Masami, additional, SETOGAWA, Tomoichi, additional, SEMBA, Reiji, additional, NOGUCHI, Tetsuya, additional, KATOU, Katsuaki, additional, SASANO, Hironobu, additional, KIKUCHI, Akihiko, additional, NAGURA, Hiroshi, additional, Tsuyama, S, additional, Yang, D-H, additional, Ohmori, J, additional, Ge, Y-B, additional, Murata, F, additional, FUJIMOTO, Toyoshi, additional, UNE, Tomoko, additional, SHIOYA, Mariko, additional, KOGO, Hiroshi, additional, YOKOTA, Sadaki, additional, KURONO, Chieko, additional, WATABIKI, Toshimitsu, additional, YOSHIDA, Manabu, additional, OKII, Yutaka, additional, YOSHIMURA, Sumitaka, additional, TOKIYASU, Takuma, additional, AKANE, Atsushi, additional, INOUE, Satoko, additional, NAITO, Ichiro, additional, SENO, Satimaru, additional, MAKIDONO, Chiho, additional, TOKUNAGA, Shoko, additional, IMAI, Shinji, additional, Kawai, Norio, additional, INOKUCHI, Tetsuo, additional, KONDO, Teruyoshi, additional, OHTA, Keisuke, additional, ANNOH, Hiromichi, additional, ISHIBASHI, Yoshihiro, additional, Yasuda, Masanori, additional, Okabe, Tsuyoshi, additional, Takekoshi, Susumu, additional, Hasegawa, Hideaki, additional, Itoh, Johbu, additional, Osamura, Yoshiyuki, additional, Watanabe, Keiichi, additional, TAIUZAWA, Toshihiro, additional, SAITO, Takuma, additional, and YASHIRO, Takashi, additional

Published
1998
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29. Abstracts

Author

Zhou, Xinhua, primary, Kudo, Akihiko, additional, Kawakami, Hayato, additional, Hirano, Hiroshi, additional, FAYED, M.H., additional, MAKITA, T., additional, SUZAKI, Etsuko, additional, KATAOKA, Katsuko, additional, Katsumata, Osamu, additional, Fujimoto, Kazushi, additional, Yamashina, Shohei, additional, USUDA, Nobuteru, additional, JOHKURA, Kohhei, additional, SUGANUMA, Tatsuo, additional, SAWAGUCHI, Akira, additional, NAGAIKE, Ryoko, additional, KAWANO, Jun-ichi, additional, OINUMA, Tsutomu, additional, Izumi, Shin-ichi, additional, Iwamoto, Michiko, additional, Shin, Masashi, additional, Nakano, Paul K., additional, Ueda, Tadashi, additional, Ishikawa, Yoshimaro, additional, Kubo, Eri, additional, Miyoshi, Norio, additional, Fukuda, Masaru, additional, Akagi, Yoshio, additional, Miki, H., additional, Nakajima, M., additional, Yuge, K., additional, Taomoto, M., additional, Tsubura, A., additional, Shikata, N., additional, Senzaki, H., additional, MASUDA, Atsushi, additional, NAGAOKA, Takanori, additional, OYAMADA, Masahito, additional, TAKAMATSU, Tetsuro, additional, Furuta, Hirokazu, additional, Hata, Yoshinobu, additional, Yokoyama, Keiichi, additional, Takamatsu, Tetsuro, additional, Itoh, J., additional, Takumi, I., additional, Kawai, K., additional, Serizawa, A., additional, Sanno, N., additional, Teramoto, A., additional, Osamura, R.Y., additional, MATSUTA, Morimasa, additional, MATSUTA, Mayumi, additional, I, Nishiya, additional, TAKAHASHI, Satoru, additional, KAWABE, Kazuki, additional, LIEBER, Michael M., additional, JENKINS, Robert B, additional, SASANO, HIRONOBU, additional, IINO, KAZUMI, additional, SUZUKI, TAKASHI, additional, NAGURA, HIROSHI, additional, Ge, Y-B, additional, Ohmori, J., additional, Tsuyama, S., additional, Yang, D-H, additional, Murata, F., additional, JOHKURA, Kohei, additional, LIANG, Yan, additional, MATSUI, Toshifumi, additional, NAKAZAWA, Ayami, additional, HIGUCHI, Susumu, additional, MATSUSHITA, Yukio, additional, Naritaka, Heiji, additional, Kameya, Toru, additional, Sato, Yuichi, additional, Inoue, Hiroshi, additional, Otani, Mitsuhiro, additional, Kawase, Takeshi, additional, KUROOKA, Yuji, additional, NASU, Kimio, additional, KAMEYAMA, Shuji, additional, MORIYAMA, Nobuo, additional, YANO, Junichi, additional, TSUJIMOTO, Gozo, additional, Matsushita, Tsutomu, additional, Oyamada, Masahito, additional, YAMAMOTO, Hitoshi, additional, MATSUURA, Junko, additional, NOMURA, Takako, additional, SASAKI, Junzo, additional, NAWA, Tokio, additional, KITAZAWA, Riko, additional, KITAZAWA, Sohei, additional, KASIMOTO, Hideyoshi, additional, MAEDA, Sakan, additional, WATANABE, Jun, additional, Mino, Kazuto, additional, KONDO, Kazumasa, additional, KANAMURA, Shinsuke, additional, Ueki, Tetsuo, additional, Takeuchi, Takumi, additional, Nishimatsu, Hiroaki, additional, Kajiwara, Takahiro, additional, Moriyama, Nobuo, additional, Kawabe, Kazuki, additional, Tominaga, Takashi, additional, Kobayashi, Ken-ichiro, additional, Minei, Sadatugu, additional, Okada, Yasuhiro, additional, Yamanaka, Yataro, additional, Ichinose, Taketo, additional, Hachiya, Takahiko, additional, Hirano, Daisaku, additional, Ishida, Hajime, additional, Okada, Kiyoki, additional, HASEGAWA, Hideaki, additional, WATANABE, Keiichi, additional, ITOH, J., additional, HASEGAWA, H., additional, UMEMURA, S., additional, YASUDA, M., additional, TAKEKOSHI, S., additional, OSAMURA, R.Y., additional, WATANABE, K., additional, TAKEDA, Kazuo, additional, HOSHI, Tatsuya, additional, KATO, Katsuaki, additional, OHARA, Shuichi, additional, KONNO, Ryo, additional, ASAKI, Shigeru, additional, TOYOTA, Takayoshi, additional, TATENO, Hiroo, additional, NISHIKAWA, Sumio, additional, SASAKI, Fumie, additional, Ito, Yuko, additional, Matsumoto, Kenji, additional, Daikoku, Eriko, additional, Otsuki, Yoshinori, additional, SANO, Makoto, additional, UMEZAWA, Akihiro, additional, ABE, Hitoshi, additional, FUKUMA, Mariko, additional, SUZUKI, Atsushi, additional, ANDO, Takashi, additional, and HATA, Jun-ichi, additional

Published
1998
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36. GENERAL SESSION

Author

Aida, Takeo, primary, Urata, Shinichi, additional, Oquro, Tatsuo, additional, AKIMOTO, Yoshihiro, additional, OBINATA, Akiko, additional, ODA, Yuko, additional, ENDO, Hiroyoshi, additional, KASAI, Ken-ichi, additional, HIRANO, Hiroshi, additional, AOKI, Toshihiro, additional, OINUMA, Tsutomu, additional, KAWANO, Junichi, additional, SUGANUMA, Tatsuo, additional, Arai, Ryohachi, additional, Kojima, Yasuji, additional, Maeda, Toshihiro, additional, Arakawa, M., additional, Mizoguchi, A., additional, Fujimoto, E., additional, Miki, A., additional, Ide, C., additional, ARAKI, Nobukazu, additional, TAKASHIMA, Yoichiro, additional, OGAWA, Kazuo, additional, ARAKI, Tsutomu, additional, Chang, C.K., additional, Ogawa, Y., additional, Kuwahara, H., additional, Yagi, T., additional, CHIDA, Kohsuke, additional, CHIKAMORI, Kensuke, additional, CUI, Yong-xi, additional, KIGUCHI, Kazushige, additional, NOZAWA, Shirou, additional, IWAMORI, Masao, additional, NAGAI, Yoshitaka, additional, KAWAKAMI, Hayato, additional, DAIMON, T., additional, KAWAI, K., additional, UCHIDA, K., additional, DATE, Fumiko, additional, SASANO, Hironobu, additional, NAGURA, Hiroshi, additional, DOBASHI, Kazushige, additional, MUNIM, Afreen, additional, ASAYAMA, Kohtaro, additional, SUZUKI, Koichi, additional, KATO, Kiyohiko, additional, KAWAOI, Akira, additional, ENDO, Hisashi, additional, YAMADA, Gotaro, additional, NISHIMOTO, Hiroshi, additional, TSUJI, Takao, additional, NAKANE, Paul K., additional, FUJII, Tetsuya, additional, KOMORI, Kaoru, additional, SAKAI, Masao, additional, YAMADA, Keiki, additional, KARASAWA, Nobuyuki, additional, NAGATSU, Ikuko, additional, FUJIMOTO, Toyoshi, additional, NAKADE, Shinji, additional, MIKOSHIBA, Katsuhiko, additional, FUKUDA, Kuniaki, additional, FUKAMI, Masaharu, additional, FUKUI, Makoto, additional, KIMURA, Kumiko, additional, QIAO, Yan, additional, MURATA, Junzo, additional, ASANO, Goro, additional, HANAI, Toru, additional, USUDA, Nobuteru, additional, MORITA, Takashi, additional, KONG, Yonuli, additional, NAGATA, Tetsuji, additional, HARADA, Takayuki, additional, HASHIMOTO, Koichi, additional, TORII, Ikuko, additional, MORIKAWA, Shigeru, additional, HAREYAMA, Masashi, additional, FUJIMOTO, Kazushi, additional, HONDA, Yoshihito, additional, HIRAKAWA, Makoto, additional, KAWATA, Mitsuhiro, additional, HIROSE, Yayoi, additional, WATANABE, Masahito, additional, SHIMADA, Masahisa, additional, IHIDA, K., additional, TSUYAMA, S., additional, NASHIO, N., additional, MURATA, F., additional, KATSUYAMA, T., additional, OHTA, H., additional, IMAMOTO, Kikuko, additional, NAGATU, Ikuko, additional, Inada, K., additional, Utunomiya, H., additional, Sato, K., additional, Osamura, R.Y., additional, Katakami, H., additional, Mayo, K., additional, ISHIBASHI, Toshimitsu, additional, KANAZAWA, Kyotaro, additional, SAITO, Takuma, additional, ITO, Nobuaki, additional, KAWAHARA, Shingo, additional, HIRANO, Yoshinari, additional, HIROTA, Tadaomi, additional, ITOH, Atsuko, additional, ITOH, Kinji, additional, TAKESHITA, Tomoko, additional, ITOH, Masamichi, additional, Itoh, J., additional, Watanabe, K., additional, Itoh, Y., additional, Komatsu, N., additional, Angeletti, H., additional, ITO-SAITO, Atsuko, additional, SAITO, Naoaki, additional, MATSUMURA, Takeo, additional, TANAKA, Chikako, additional, IWAMOTO, Masunobu, additional, WATANABE, Jun, additional, ASADA-KUBOTA, Mari, additional, and KANAMURA, Shinsuke, additional

Published
1991
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40. Expression of foreign type I ribulose-1,5-bisphosphate carboxylase/ oxygenase (EC 4.1.1.39) stimulates photosynthesis in cyanobacterium SynechococcusPCC7942 cells

Author

Iwaki, T., Haranoh, K., Inoue, N., Kojima, K., Satoh, R., Nishino, T., Wada, S., Ihara, H., Tsuyama, S., Kobayashi, H., and Wadano, A.

Abstract

A reporter gene assay revealed that promoters derived from SynechococcusPCC7942 (S.7942) psbAIand SynechocystisPCC6803 (S.6803) psbAII were suitable for the expression of foreign ribulose-bisphosphate carboxylase (RuBisCO; EC 4.1.1.39) in S.7942cells. Transformational vectors with a promoter and a foreign RuBisCO gene, cvrbcoriginated from Allochromatium vinosum, were constructed on a binary vector, pUC303, and introduced to S.7942cells. When the cvrbcwas expressed with the S.7942 psbAI promoter, the total RuBisCO activity increased 2.5- to 4-fold than that of the wild type cell. The S.6803 psbAII promoter increased the activity of the transformant 1.5–2 times of that of wild type cell. There was a significant increase in the rate of photosynthesis depending on the increase of RuBisCO activity. The maximum rate of photosynthesis of the transformant cell was 1.63 times higher than that of the wild type under the illumination of 400 μmol m−2s−1, at 20 mM bicarbonate and at 30 °C. Although the photosynthesis of the higher plant is limited by the ability of photosystems under high irradiance and the high CO2 concentration, that of the S.7942cell is limited by the RuBisCO activity, even at high CO2concentrations and under high irradiance.A reporter gene assay revealed that promoters derived from SynechococcusPCC7942 (S.7942) psbAIand SynechocystisPCC6803 (S.6803) psbAII were suitable for the expression of foreign ribulose-bisphosphate carboxylase (RuBisCO; EC 4.1.1.39) in S.7942cells. Transformational vectors with a promoter and a foreign RuBisCO gene, cvrbcoriginated from Allochromatium vinosum, were constructed on a binary vector, pUC303, and introduced to S.7942cells. When the cvrbcwas expressed with the S.7942 psbAI promoter, the total RuBisCO activity increased 2.5- to 4-fold than that of the wild type cell. The S.6803 psbAII promoter increased the activity of the transformant 1.5–2 times of that of wild type cell. There was a significant increase in the rate of photosynthesis depending on the increase of RuBisCO activity. The maximum rate of photosynthesis of the transformant cell was 1.63 times higher than that of the wild type under the illumination of 400 μmol m−2s−1, at 20 mM bicarbonate and at 30 °C. Although the photosynthesis of the higher plant is limited by the ability of photosystems under high irradiance and the high CO2 concentration, that of the S.7942cell is limited by the RuBisCO activity, even at high CO2concentrations and under high irradiance.

Published
2006
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44. Postembedding staining of Brunner's gland with lectin-ferritin conjugates.

Author

Suzuki, S, Tsuyama, S, Suganuma, T, Yamamoto, N, and Murata, F

Abstract

Postembedding staining of intracellular carbohydrates of rat Brunner's gland cells embedded in Epon and acrylamide was carried out with Ricinus communis agglutinin-ferritin, concanavalin A-ferritin, and wheat germ agglutinin-ferritin conjugates. Th Golgi vacuoles and mucous granules were stained with these conjugates. In each staining, the tissues embedded in acrylamide were stained more strongly than those embedded in Epon. The staining intensity of wheat germ agglutinin-ferritin was the strongest among the three conjugates and the staining intensity of Ricinus communis agglutinin-ferritin was stronger than that of concanavalin A-ferritin in both embedding methods. Free ferritin showed almost no binding to these structures and staining with the conjugates was inhibited by the addition of appropriate competitive sugars to the staining solutions. Osmium-postfixed tissues were not stained well with the conjugates. Washing of the sections with bovine serum albumin solution after staining was an essential step in the present method to reduce the nonspecific adsorption of the conjugates. The present method was very simple and had good reproducibility.

Published
1981
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45. Distribution of Glycoconjugates in the Kidney Studied by Use of Labeled Lectins1

Author

Murata, F., Tsuyama, S., Suzuki, S., Hamada, H., Ozawa, M., and Muramatsu, T.

Abstract

Distribution of glycoconjugates in different areas of the rat kidney was studied by light and electron microscopy using six different horseradish peroxidase-labeled lectins. Glomeruli and brush borders of the proximal tubules reacted differently to these lectins, which indicated differences in the carbohydrate compositions of those regions. The ascending limb of Henle's loop (ALH) had strong binding sites for peanut agglutinin (PNA) and soybean agglutinin (SBA). Dolichos biflorus agglutinin (DBA) did not stain the cells of ALH but did stain those of distal convoluted tubules (DCT). DBA is a good marker for distinguishing ALH from DCT. DBA, PNA, and SBA were also good markers of the collecting duct. Ricinus communis agglutinin (RCA-1) and wheat germ agglutinin (WGA) diffusely stained the various components of different parts of the kidney.

Published
1983
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48. Purification, characterization, and localization of an ADP-ribosylactin hydrolase that uses ADP-ribosylated actin from rat brains as a substrate.

Author

Okamoto, H, Fujita, H, Matsuyama, S, and Tsuyama, S

Abstract

Mammalian ADP-ribosylation is poorly understood. An ADP-ribosylprotein hydrolase that acted on ADP-ribosylated actin was purified from rat brain. The molecular weight of this enzyme was 62, 000 as determined by SDS-polyacrylamide gel electrophoresis and gel filtration. Enzyme activity with ADP-ribosylated actin as a substrate was inhibited by NAD, ATP, ADP, and ADP-ribose, but not by AMP. Mg2+ increased Vmax. Purified ADP-ribosylactin hydrolase catalyzed the hydrolysis of ADP-ribosylated subunits Gsalpha, Gialpha, and Goalpha and elongation factor-2. After de-ADP-ribosylation by the purified ADP-ribosylactin hydrolase, the proteins were re-ADP-ribosylated by brain mono-ADP-ribosyltransferases and bacterial toxins. The actin that was de-modified by ADP-ribosylactin hydrolase could form actin filaments. Two kinds of monoclonal antibodies against ADP-ribosylactin hydrolase were prepared and characterized. In an immunohistochemical study, the plasma membranes and cytoplasmic regions of the nerve cells in the rat brain were immunoreactive. In subcellular fractionation of the brains, most of the ADP-ribosylactin hydrolase activity was found in the cytosol and synaptosome fractions. When the synaptosomes were treated with a hypotonic solution, ADP-ribosylactin hydrolase activity was found in the supernatant. Our findings suggest that brain ADP-ribosylactin hydrolase has the important function of polymerizing actin for signal transduction in the cytosol of nerve cells and synaptosomes.

Published
1997

49. Numerous phosphates of microtubule-associated protein 2 in living rat brain.

Author

Tsuyama, S, Terayama, Y, and Matsuyama, S

Abstract

Microtubule-associated protein 2 (MAP 2) purified from microwave-irradiated rat head contained about 46 esterified phosphates (mole/mol), which were not bound covalently to lipids and did not assemble with microtubules. After some phosphates were released by calf intestinal alkaline phosphatase, the phosphate content of MAP-2 decreased to 16 mol of phosphate and the protein assembled in vitro. MAP-2 purified after microtubule assembly cycles and also the cytosolic heat-stable fraction without assembly cycles had 10 mol of phosphate, and both assembled with microtubules. The MAP-2 with 46 phosphates and that with 10 had different pI in isoelectric focusing, but the components, MAP-2a and −2b, were always near each other. In high-pressure liquid chromatography, MAP-2 containing 46 mol of phosphate appeared after that 10 mol of phosphate. Phosphoserine, phosphothreonine, and phosphotyrosine were recovered from tryptic digestion of MAP-2 with 46 mol of phosphate. These findings suggest that two kinds of MAP-2, one with 46 phosphates and not bound to tubulin and the other with 10-16 phosphates and bound to tubulin, are present in the living rat brain.

Published
1987
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50. Calcium/phospholipid-dependent kinase recognizes sites in microtubule-associated protein 2 which are phosphorylated in living brain and are not accessible to other kinases.

Author

Tsuyama, S, Bramblett, G T, Huang, K P, and Flavin, M

Abstract

Microtubule-associated protein 2 (MAP-2) purified after microtubule assembly cycles from bovine brain had been shown to contain about 10 esterified phosphates (mol/mol), which were relatively phosphatase resistant and essentially confined to the projection domain which contributes to the visible arms on microtubules. The kinase responsible for phosphorylating these sites had not been identified. We have approached this question by using a phosphatase that releases the bulk of these residues and then determining which kinase can now add additional residues corresponding to those released. Three kinases were chosen because of their abundance in brain and/or proximity to microtubules. Of these only Ca/phospholipid-dependent kinase was able to recognize the previously occupied sites. We also found that MAP-2 isolated from rat brain without assembly cycles contained more phosphate than previously recognized, greater than 30 mol/mol, suggesting that 20 of these had been inadvertently released by phosphatase during assembly cycles. All 3 kinases (Ca/phospholipid-dependent, cAMP-dependent, and Ca/calmodulin-dependent kinase II) recognized more sites in the bovine than in the rat MAP-2.

Published
1986
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