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16. [Cl.sup.-]-dependent upregulation of human organic anion transporters: different effects on transport kinetics between hOAT1 and hOAT3

Author

Ueo, Harumasa, Motohashi, Hideyuki, Katsura, Toshiya, and Inui, Ken-ichi

Subjects
Chloride channels -- Research, Biological transport, Active -- Research, Anion exchangers (Biology) -- Research, Biological sciences
Abstract

Chloride ion has a stimulatory effect on the transport of organic anions across renal basolateral membranes. However, the exact mechanisms at molecular levels have been unclear as of yet. Human organic anion transporters hOAT1 and hOAT3 play important roles in renal basolateral membranes. In this study, the effects of C1 on the activities of these transporters were evaluated by using HEK293 cells stably expressing hOAT1 or hOAT3 (HEK-hOAT1 or HEK-hOAT3). The uptake of p-[[sup.14]C]aminohippu-rate by HEK-hOAT1 and [[sup.3]H]estrone sulfate by HEK-hOAT3 was greater in the presence of [Cl.sup.-] than in the presence of S[O.sup.2-.sub.4] or gluconate. Additionally, the uptake of various compounds by HEK-hOAT1 and HEK-hOAT3 was significantly higher in the [Cl.sup.-]-containing medium than the gluconate-containing medium, suggesting that the influences of [Cl.sup.-] are not dependent on substrate and that [Cl.sup.-] directly stimulates the functions of hOAT1 and hOAT3. The substitution of gluconate with [Cl.sup.-] did not change the [K.sub.m] value for the uptake of p-[[sup.14]C]aminohippurate by HEK-hOAT1 but caused an approximately threefold increase in the maximal uptake rate ([V.sub.max]) value. On the other hand, replacement of gluconate with C1 decreased the [K.sub.m] value for the uptake of [[sup.3]H]estrone sulfate and cefotiam by HEK-hOAT3 to about one-third, while it did not change the [V.sub.max] value. In summary, [Cl.sup.-] upregulates the activities of both hOAT1 and hOAT3, but its effects on transport kinetics differ between these transporters. It was suggested that [Cl.sup.-] participates in the trans-location process for hOAT1, and the substrate recognition process for hOAT3. renal secretion; basolateral membrane; SLC22A; HEK293 doi:10.1152/ajprenal.00376.2006

Published
2007

18. Abstracts of Poster Presentations

Author

Iguchi, Megumi, Ishiwata, Isamu, Tamagawa, Tomoharu, Ishiwata, Chieko, Kiguchi, Kazushige, Sato, Kahei, Saito, Shigeo, Yamamoto, Motoyoshi, Hashimoto, Hisashi, Tachibana, Toshiaki, Ohi, Satoshi, Akahori, Masakazu, Yokose, Takashi, Ishikawa, Hiroshi, Ohi S., Sato K., Kimura M., Kobayashi M., Hosaka K., Ishiwata, Ciko, Kiguci, Kazushige, Etsuko, Hasegawa, Tomoharu, Tamagawa, Yuko, Tokieda, Isamu, Ishiwata, Kahei, Sato, Sunaga, Junko, Tomimai, Ryoko, Motohashi, Hideyuki H., Nagaike, Koki, Tanaka, Hiroyuki, Itoh, Hiroshi, Fukushima, Tsuyoshi, Kohama, Kazuyo, Uchiyama, Shuichiro, Kataoka, Hiroaki, Yamamoto M., Yoshimoto T., Sekiya H., Negishi Y., Sakoda K., Izumi Y., Takao S., Ishikawa H., Tachibana T., Hashimoto H., Motoyama S., Sato E., Kamoi K., Iino M., Okayasu K., Osawa G., Nakaya H., Enomoto, Keiko, Kamoi, Hisahiro, Sato, Soh, Kamoi, Kyuichi, Wachi, Tomoka, Yoshida, Noriyuki, Yano, Yoshihisa, Ueno, Munehisa, Deguchi, Nobuhiro, Hirotsune, Shinji, Sasaki, Hiroyuki, Tsukita, Shoichiro, Tabei I., Ito R., Kyouda S., Ohkubo T., Ishida Y., Ishiwata I., Satou K., Kubo H., Yamazaki Y., Yanaga K., Sato, Yorino, Kazami, Ai, Okamoto, Naoki, Hosaka, Koji, Soma, Masayuki, Sato, Yoshiro, Sonobe, Makoto, Niimi S., Yamada K., Yasuda M., Tanaka T., Yamada, Kyosuke, Iida, Yasushi, Ueda, Kazu, Misawa, Akihiko, Suzuki, Nagazumi, Takahashi, Hiroyuki, Kato, Hiroyuki, Kimura, Eizo, Yasuda, Makoto, Tanaka, Tadao, Hiroshi, Mizuhara, Yohichi, Kobayashi, Yosuke, Kanishi, Soujiro, Sato, Haruhiro, Kondo, Yoshiki, Ohkuma, Tazuru, Ohhara, Yoshiko, Okuda, Mari, Watanabe, Asami, Okamura, Tomohiro, Iida, Kazushige, Kiguchi, Bunpei, Ishizuka, Ishikawa, Mayumi, Kimura, Kazuhiro, Akishima, Yuri, Shimojo, Masako, Ueshiba, Hajime, Tsuboi, Kumiko, Nakata, Koichiro, Akasaka, Yoshikjo, Shibuya, Kazutoshi, Yoshino, Gen, Matsuo, Yoshinobu, Drexler, Hans G., Harashima, Akira, Okochi, Ayumi, Orita, Kunzo, Kashiwagi H., Shigeya, Kyouda, Isao, Tabei, Satoshi, Ohi, Toshiaki, Tachibana, Hirotaka, Kubo, Hisashi, Hashimoto, Katsuhiko, Yanaga, Yoji, Yamazaki, Hiroshi, Ishikawa, Kudoh K., Takano M., Okamoto S., Sasaki K., Fujii K., Hirata J., Kita T., Kikuchi Y., Yoshioka, Kunihiko, Ohno, Yoshio, Gondo, Tatsuo, Nakagami, Yoshihiro, Matsushita, Kazuhito, Sakamoto, Noboru, Matumoto, Mayuko, Koga, Shouji, Namiki, Kazunori, Aoyagi, Teiichirou, Ohori, Makoto, Hatano, Tadashi, Tachibana, Masaaki, Kawamura, Takeshi, Nishimura, Toshihide, Mori, Y., Okumura, T., Shimada, Y., Okabe, Michio, Kaganoi, Junichi, Nagatani, Shiro, Kondo, Masato, Shimada, Yutaka, Watanabe, Go, Imamura, Masayuki, Kitahara, Shinzo, Tagawa, Yutaka, Ita, Masamichi, Smolewski, Piotr, Halicka, Dorota, and Darzynkiewicz, Zbigniew

Published
2004
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19. Abstracts of Oral Presentations

Author

Tamagawa, Tomoharu, Ishiwata, Isamu, Saito, Shigeo, Sakaguchi E., Iwasawa M., Sato K., Motohashi, Hideyuki H., Kada, Hidemi, Sato, Kahei, Yano, Yoshihisa, Yoshida, Noriyuki, Wachi, Tomoka, Ueno, Munehisa, Deguchi, Nobuhiro, Hirotsune, Shinji, Hara, Satoshi, Oya, Mototsugu, Mizuno, Ryuichi, Horiguchl, Akio, Marumo, Ken, Murai, Masaru, Ikegami, Shusei, Tadakuma, Takushi, Suzuki, Satoshi, Asano, Tomohiko, Hayakawa, Masamichi, Iino, Satoshi, Abeyama, Kazuhiro, Takao, Sonshin, Aikou, Takashi, Maruyama, Dairo, Kobayashi, Yukiko, Ohtsuki, Mamitaro, Sutheesophon, Krittaya, Ishii, Hideshi, Kitayama, Hitoshi, Kano, Yasuhiko, Nakagawa, Hidemi, Furukawa, Yusuke, Li, Kyo, Sakusai, Yasuo, Ryu, Takashi, Aduma, Keiichi, Kasai, Harubumi, Numa, Yoshihiro, Inagaki, Takayuki, Yamanouchi, Yasuo, Kawamoto, Keiji, Nawashiro, Hiroshi, Nomura, Namiko, Ooigawa, Hidetoshi, Otani, Naoki, Shinomiya, Nariyoshi, and Matsuo, Hirotaka

Published
2004
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24. Thyroid hormone regulates the activity and expression of the peptide transporter PEPT1 in Caco-2 cells

Author

Ashida, Kayoko, Katsura, Toshiya, Motohashi, Hideyuki, Saito, Hideyuki, and Inui, Ken-Ichi

Subjects
Physiology -- Research, Thyroid hormones -- Physiological aspects, Gene expression -- Physiological aspects, Intestinal absorption -- Genetic aspects, Hormones -- Physiological aspects, Biological sciences
Abstract

An oligopeptide transporter (PEPT1) in the small intestine plays an important role in the absorption of small peptides and peptide-like drugs. We examined the effect of thyroid hormone 3,5,3'-L-triiodothyronine ([T.sub.3]) on the activity and expression of PEPT1 in human intestinal Caco-2 cells. Treatment of Caco-2 cells with [T.sub.3] inhibited [[sup.14]C]glycylsarcosine uptake in a time- and dose-dependent manner. [sup.14]C]glycylsarcosine uptake was reduced by pretreatment of the cells with 100 nM [T.sub.3] for 4 days (67% of control value), whereas methyl-[alpha]-D-[U-[sup.14]C]glucopyranoside and [[sup.3]H]threonine uptake were not decreased. Kinetic analysis showed that [T.sub.3] treatment significantly decreased the maximum uptake ([V.sub.max]) value for [[sup.14]C]glycylsarcosine uptake but had no effect on the [K.sub.m] value. Moreover, [T.sub.3] treatment caused a significant decrease in the amount of PEPT1 mRNA (25% of the control). Western blotting indicated that the amount of PEPT1 protein in the apical membrane was decreased (70% of the control). These findings indicate that [T.sub.3] treatment inhibits the uptake of [[sup.14]C]glycylsarcosine by decreasing the transcription and/or stability of PEPT1 mRNA. intestinal absorption; intestinal oligopeptide transporter; hormonal regulation; gene expression

Published
2002

34. Transgenic Monkey Model of the Polyglutamine Diseases Recapitulating Progressive Neurological Symptoms

Author

Tomioka, Ikuo, primary, Ishibashi, Hidetoshi, additional, Minakawa, Eiko N., additional, Motohashi, Hideyuki H., additional, Takayama, Osamu, additional, Saito, Yuko, additional, Popiel, H. Akiko, additional, Puentes, Sandra, additional, Owari, Kensuke, additional, Nakatani, Terumi, additional, Nogami, Naotake, additional, Yamamoto, Kazuhiro, additional, Noguchi, Satoru, additional, Yonekawa, Takahiro, additional, Tanaka, Yoko, additional, Fujita, Naoko, additional, Suzuki, Hikaru, additional, Kikuchi, Hisae, additional, Aizawa, Shu, additional, Nagano, Seiichi, additional, Yamada, Daisuke, additional, Nishino, Ichizo, additional, Ichinohe, Noritaka, additional, Wada, Keiji, additional, Kohsaka, Shinichi, additional, Nagai, Yoshitaka, additional, and Seki, Kazuhiko, additional

Published
2017
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44. Live, full-term mouse pups from oocytes grown and matured in vitrowith serum substitutes

Author

Motohashi, Hideyuki H., Taniguchi, Ryoma, Sakamoto, Junpei, Sankai, Tadashi, and Kada, Hidemi

Abstract

For in vitrogrowth and maturation of mouse oocytes (IVG-IVM), serum is added to media up to and including the stage of oocyte maturation; this subsequently supports oocytes through fertilization and early embryo development. However, problems may occur with sera, such as batch differences and issues of biosafety. The purpose of the present study was to determine the capacity for fertilization and pre- and post-implantation development of oocytes that underwent IVG-IVM with a serum substitute. Oocyte-granulosa cell complexes from preantral follicles were cultured in medium with either fetal bovine serum (FBS), Serum Substitute Supplement™ (SSS), or Knockout™ Serum Replacement (KSR) for 10days, and were then allowed to mature for 17 h. Subsequently, more than 90% of oocytes underwent germinal vesicle breakdown (GVBD) and more than 70% reached metaphase II, with no significant difference between the groups. A lower fertilization rate, presumably due to zona hardening, was found in the serum substitute groups. Nevertheless, more than 50% of the inseminated oocytes were fertilized and 35%–45% of them underwent first cleavage and developed to the blastocyst stage. Following embryo transfer, one and four live offspring were produced from the SSS and KSR groups, respectively. The present study demonstrated that murine IVG-IVM oocytes cultured in media with a serum substitute, achieved fertilization in vitro, pre- and post-implantation development, and the delivery of live pups, although the efficiency of the process is reduced compared to FBS supplementation.

Published
2017
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