Faculty Profiles - TOYOKUNI Shinya

Papers - TOYOKUNI Shinya

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Functional assessment of self-renewal activity of male germline stem cells following cytotoxic damage and serial transplantation. Reviewed International journal

Mito Kanatsu-Shinohara, Shinya Toyokuni, Takeshi Morimoto, Shigeyuki Matsui, Tasuku Honjo, Takashi Shinohara

Biology of reproduction Vol. 68 ( 5 ) page： 1801 - 1807 2003.5

Language：English Publishing type：Research paper (scientific journal)

Spermatogenesis is dependent on a small population of stem cells. Although stem cells are believed to expand infinitely, there is little functional evidence regarding whether spermatogonial stem cells can increase in their number. Using the spermatogonial transplantation technique, we evaluated the proliferative potential of spermatogonial stem cells in two models of regeneration. After busulfan injection to deplete stem cells, the surviving stem cells were able to expand by at least 15.8-fold within 2 mo. On the other hand, a serial transplantation study indicated that one transplanted stem cell was able to expand by 3.8- and 12-fold within 2 and 4 mo, respectively. These results provide direct functional evidence for the expansion of stem cells and establish the basis for further characterization of the stem cell self-renewal process.

DOI： 10.1095/biolreprod.102.012575

Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis

TOYOKUNI Shinya, KAWAGUCHI Waka, AKATSUKA Shinya, HIROYASU Makoto, HIAI Hiroshi

Pathol Int Vol. 53 ( 4 ) page： 259 - 261 2003.4

Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis

TOYOKUNI Shinya, KAWAGUCHI Waka, AKATSUKA Shinya, HIROYASU Makoto, HIAI Hiroshi

Pathology international Vol. 53 ( 4 ) page： 259 - 261 2003.4

Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis

Toyokuni S, Kawaguchi W, Akatsuka S, Hiroyasu M, Hiai H

PATHOLOGY INTERNATIONAL Vol. 53 ( 4 ) page： 259 - 261 2003.4

Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis.

Toyokuni S, Kawaguchi W, Akatsuka S, Hiroyasu M, Hiai H

Pathology international Vol. 53 ( 4 ) page： 259 - 61 2003.4

Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis. International journal

Shinya Toyokuni, Waka Kawaguchi, Shinya Akatsuka, Makoto Hiroyasu, Hiroshi Hiai

Pathology international Vol. 53 ( 4 ) page： 259 - 61 2003.4

Glycine ameriorates lung reprefusion injury after cold preservation in an ex vivo rat lung model.

Omasa M, Fukuse T, Toyokuni S, Mizutani Y, Yoshida H, Ikeyama K, Hasegawa S, Wada H

TRANSPLANTATION Vol. 75 ( 5 ) page： 591 - 598 2003.3

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Transplantation

Background. The role of glycine has not been investigated in lung ischemia-reperfusion injury after cold preservation. Furthermore, the role of apoptosis after reperfusion following cold preservation has not been fully understood. Methods. Lewis rats were divided into three groups (n=6 each). In the GLY(-) and GLY(+) groups, isolated lungs were preserved for 15 hr at 4°C after a pulmonary artery (PA) flush using our previously developed preservation solution (ET-K; extracellular-type trehalose containing Kyoto), with or without the addition of glycine (5 mM). In the Fresh group, isolated lungs were reperfused immediately after a PA flush with ET-K. They were reperfused for 60 min with an ex vivo perfusion model. Pulmonary function, oxidative stress, apoptosis, and tumor necrosis factor (TNF)-α expression were assessed after reperfusion. Results. Shunt fraction and peak inspiratory pressure after reperfusion in the GLY(-) group were significantly higher than those in the GLY(+) and Fresh groups. Oxidative damage and apoptosis in the alveolar epithelial cells of the GLY(-) group, assessed by immunohistochemical staining and quantification of 8-hydroxy-2′-deoxyguanosine and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method, were significantly higher than those of the GLY(+) and Fresh groups. There were correlations among shunt fraction, oxidative damage, and apoptosis. There was no expression of TNF-α messenger RNA in all groups evaluated by the reverse transcription-polymerase chain reaction. Conclusions. Glycine attenuates ischemia/reperfusion injury after cold preservation by reducing oxidative damage and suppressing apoptosis independent of TNF-α in this model. The suppression of apoptosis might ameliorate lung function after reperfusion.

DOI： 10.1097/01.TP.0000053200.98125.14

Web of Science

Scopus

PubMed

Glycine ameliorates lung reperfusion injury after cold preservation in an ex vivo rat lung model. International journal

Mitsugu Omasa, Tatsuo Fukuse, Shinya Toyokuni, Yoichi Mizutani, Hiroshi Yoshida, Kazuyuki Ikeyama, Seiki Hasegawa, Hiromi Wada

Transplantation Vol. 75 ( 5 ) page： 591 - 8 2003.3

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: The role of glycine has not been investigated in lung ischemia-reperfusion injury after cold preservation. Furthermore, the role of apoptosis after reperfusion following cold preservation has not been fully understood. METHODS: Lewis rats were divided into three groups (n=6 each). In the GLY(-) and GLY(+) groups, isolated lungs were preserved for 15 hr at 4 degrees C after a pulmonary artery (PA) flush using our previously developed preservation solution (ET-K; extracellular-type trehalose containing Kyoto), with or without the addition of glycine (5 mM). In the Fresh group, isolated lungs were reperfused immediately after a PA flush with ET-K. They were reperfused for 60 min with an ex vivo perfusion model. Pulmonary function, oxidative stress, apoptosis, and tumor necrosis factor (TNF)-alpha expression were assessed after reperfusion. RESULTS: Shunt fraction and peak inspiratory pressure after reperfusion in the GLY(-) group were significantly higher than those in the GLY(+) and Fresh groups. Oxidative damage and apoptosis in the alveolar epithelial cells of the GLY(-) group, assessed by immunohistochemical staining and quantification of 8-hydroxy-2'-deoxyguanosine and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method, were significantly higher than those of the GLY(+) and Fresh groups. There were correlations among shunt fraction, oxidative damage, and apoptosis. There was no expression of TNF-alpha messenger RNA in all groups evaluated by the reverse transcription-polymerase chain reaction. CONCLUSIONS: Glycine attenuates ischemia/reperfusion injury after cold preservation by reducing oxidative damage and suppressing apoptosis independent of TNF-alpha in this model. The suppression of apoptosis might ameliorate lung function after reperfusion.

PubMed

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Tumor initiating activity of Helicobacter pylori water extract on mouse skin carcinogenesis. Reviewed International journal

Takeshi Ishikawa, Norimasa Yoshida, Harukuni Tokuda, Eiichiro Ichiishi, Masashi Kuchide, Satoshi Kokura, Yuji Naito, Shinya Toyokuni, Hoyoku Nishino, Toshikazu Yoshikawa

Cancer letters Vol. 191 ( 1 ) page： 41 - 47 2003.2

Language：English Publishing type：Research paper (scientific journal)

Helicobacter pylori (H. pylori) infection has been associated with gastric carcinogenesis, but responsible and detail mechanisms are insufficient by the absence of adequate data. To obtain direct evidence regarding the carcinogenicity of H. pylori, we investigated the initiating and promoting activity of H. pylori water extract (HPE) in two-stage mouse skin carcinogenesis model. HPE treatment, as an initiation, significantly enhanced tumor formation compared with control group. Moreover, HPE treatment increased production of 8-hydroxydeoxyguanosine in epidermal cells and HPE-initiated/TPA-promoted papillomas demonstrated a point mutation of the Ha-ras gene. These results suggest an initiating activity of HPE on two-stage mouse skin carcinogenesis.

DOI： 10.1016/S0304-3835(02)00628-6

Structural basis of protein-bound endogenous aldehydes - Chemical and immunochemical characterizations of configurational isomers of a 4-hydroxy-2-nonenal-histidine adduct International journal

Hashimoto, M; Sibata, T; Wasada, H; Toyokuni, S; Uchida, K

JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 278 ( 7 ) page： 5044 - 5051 2003.2

Language：English Publishing type：Research paper (scientific journal) Publisher：Journal of Biological Chemistry

4-Hydroxy-2-nonenal (HNE), a major racemic product of lipid peroxidation, reacts with histidine to form a stable HNE-histidine Michael addition-type adduct possessing three chiral centers in the cyclic hemiacetal structure. In the present study, we characterized configurational isomers of a HNE-Nα-acetylhistidine adduct by NMR spectroscopy and by molecular orbital calculations. In addition, we raised monoclonal antibodies against (R)-HNE-histidine and (S)-HNE-histidine adducts, characterized their specificities, and examined in vivo localizations of each adduct under oxidative stress. To facilitate structural characterization of the configurational isomers of an HNE-histidine adduct, we prepared the (R)-HNE-histidine and (S)-HNE-histidine adducts by incubating Nα-acetylhistidine with each HNE enantiomer, both of which provided two peaks (Ra and Rb from (R)-HNE-histidine and Sa and Sb from (S)-HNE-histidine adducts) in reversed-phase high-performance liquid chromatography. The NMR analysis showed that each peak was a mixture of two diastereomers. In addition, the analysis of the nuclear Overhauser effect enabled the determination of configurations of the eight isomers. The relative amounts of these isomers in the NMR analysis correlated with the relative energies calculated by molecular orbital methods. On the other hand, using (R)-HNE-modified and (S)-HNE-modified keyhole limpet hemocyanins as the antigens, we raised the monoclonal antibodies, mAbR310 and mAbS412, which enantioselectively recognized the (R)-HNE-histidine and (S)-HNE-histidine adducts, respectively. Among the mixtures (Ra, Rb, Sa, and Sb) of diastereomers, mAbR310 showed the highest immunoreactivity to Rb (the mixture of 2R,4S,5R and 2S,4S,5R isomers), whereas mAbS412 preferentially recognized Sa (the mixture of 2R,4S,5S and 2S,4S,5S isomers). The presence of (R)-HNE and (S)-HNE epitopes in vivo was immunohistochemically examined in the kidney of rats exposed to the renal carcinogen, ferric nitrilotriacetate, by which nuclear and cytosolic stainings with mAbR310 and mAbS412, respectively, were detected.

DOI： 10.1074/jbc.M210129200

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PubMed

Regulation of marginal zone B cell development by MINT, a suppressor of Notch/RBP-J signaling pathway.

Kuroda K, Han H, Tani S, Tanigaki K, Tun T, Furukawa T, Taniguchi Y, Kurooka H, Hamada Y, Toyokuni S, Honjo T

Immunity Vol. 18 ( 2 ) page： 301 - 12 2003.2

Language：English Publishing type：Research paper (scientific journal) Publisher：Immunity

We found that Msx2-interacting nuclear target protein (MINT) competed with the intracellular region of Notch for binding to a DNA binding protein RBP-J and suppressed the transactivation activity of Notch signaling. Although MINT null mutant mice were embryonic lethal, MINT-deficient splenic B cells differentiated about three times more efficiently into marginal zone B cells with a concomitant reduction of follicular B cells. MINT is expressed in a cell-specific manner: high in follicular B cells and low in marginal zone B cells. Since Notch signaling directs differentiation of marginal zone B lymphocytes and suppresses that of follicular B lymphocytes in mouse spleen, the results indicate that high levels of MINT negatively regulate Notch signaling and block differentiation of precursor B cells into marginal zone B cells. MINT may serve as a functional homolog of Drosophila Hairless.

DOI： 10.1016/s1074-7613(03)00029-3

Scopus

PubMed

Regulation of marginal zone B cell development by MINT, a suppressor of Notch/RBP-J signaling pathway

Kuroda K, Han H, Tani S, Tanigaki K, Tun T, Furukawa T, Taniguchi Y, Kurooka H, Hamada Y, Toyokuni S, Honjo T

IMMUNITY Vol. 18 ( 2 ) page： 301 - 312 2003.2

Regulation of marginal zone B cell development by MINT, a suppressor of Notch/RBP-J signaling pathway. International journal

Kazuki Kuroda, Hua Han, Shoichi Tani, Kenji Tanigaki, Tin Tun, Takahisa Furukawa, Yoshihito Taniguchi, Hisanori Kurooka, Yoshio Hamada, Shinya Toyokuni, Tasuku Honjo

Immunity Vol. 18 ( 2 ) page： 301 - 12 2003.2

Language：English Publishing type：Research paper (scientific journal)

PubMed

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Astaxanthin limits exercise-induced skeletal and cardiac muscle damage in mice. Reviewed International journal

Wataru Aoi, Yuji Naito, Kunihiro Sakuma, Masashi Kuchide, Harukuni Tokuda, Takashi Maoka, Shinya Toyokuni, Shigenori Oka, Masahiro Yasuhara, Toshikazu Yoshikawa

Antioxidants & redox signaling Vol. 5 ( 1 ) page： 139 - 44 2003.2

Language：English Publishing type：Research paper (scientific journal)

Dietary antioxidants may attenuate oxidative damage from strenuous exercise in various tissues. Beneficial effects of the antioxidant astaxanthin have been demonstrated in vitro, but not yet in vivo. We investigated the effect of dietary supplementation with astaxanthin on oxidative damage induced by strenuous exercise in mouse gastrocnemius and heart. C57BL/6 mice (7 weeks old) were divided into groups: rested control, intense exercise, and exercise with astaxanthin supplementation. After 3 weeks of exercise acclimation, both exercise groups ran on a treadmill at 28 m/min until exhaustion. Exercise-increased 4-hydroxy-2-nonenal-modified protein and 8-hydroxy-2'-deoxyguanosine in gastrocnemius and heart were blunted in the astaxanthin group. Increases in plasma creatine kinase activity, and in myeloperoxidase activity in gastrocnemius and heart, also were lessened by astaxanthin. Astaxanthin showed accumulation in gastrocnemius and heart from the 3 week supplementation. Astaxanthin can attenuate exercise-induced damage in mouse skeletal muscle and heart, including an associated neutrophil infiltration that induces further damage.

DOI： 10.1089/152308603321223630

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Astaxanthin limits exercise-induced skeletal and cardiac muscle damage in mice

Aoi W, Naito Y, Sakuma K, Kuchide M, Tokuda H, Maoka T, Toyokuni S, Oka S, Yasuhara M, Yoshikawa T

ANTIOXIDANTS & REDOX SIGNALING Vol. 5 ( 1 ) page： 139 - 144 2003.2

Allogeneic offspring produced by male germ line stem cell transplantation into infertile mouse testis. Reviewed International journal

Mito Kanatsu-Shinohara, Narumi Ogonuki, Kimiko Inoue, Atsuo Ogura, Shinya Toyokuni, Tasuku Honjo, Takashi Shinohara

Biology of reproduction Vol. 68 ( 1 ) page： 167 - 173 2003.1

Language：English Publishing type：Research paper (scientific journal) Publisher：SOC STUDY REPRODUCTION

The testis is one of several immune-privileged organs and is known for its unique ability to support allogeneic or xenogeneic tissue transplants. We investigated the possibility of deriving offspring from mice that underwent transplantation with allogeneic male germ line stem cells in the testis. Although mature adult mice rejected allogeneic germ cells and were infertile, offspring were obtained by intracytoplasmic germ cell injection using partially differentiated donor cells. In contrast, complete spermatogenesis occurred when allogeneic germ cells were transplanted into immature pup testes. Tolerance induction by monoclonal antibody administration allowed the pup transplant recipients to produce allogeneic offspring by natural mating, whereas no spermatozoa were found in the epididymis of untreated recipients. Thus, these results, indicate that a histoincompatible recipient can serve as a "surrogate father" to propagate the genetic information of heterologous male donors.

DOI： 10.1095/biolreprod.102.008516

Birth of offspring following transplantation of cyropreserved immature testicular pieces and in-vitro microinsemination

Shinohara T., Inoue K., Ogonuki N., Kanatsu-Shinohara M., Miki H., Nakata K., Kurome M., Nagashima H., Toyokuni S., Kogishi K., Honjo T., Ogura A.

Human Reproduction Vol. 17 ( 12 ) page： 3039 - 3045 2002.12

Language：Japanese Publishing type：Research paper (scientific journal) Publisher：Human Reproduction

Background: Fertility protection is an urgent clinical problem for prepubertal male oncology patients who undergo either chemotherapy or radiotherapy. As these patients do not have mature sperm to be frozen, there is as yet no effective method to preserve their fertility. Methods and results: Single pieces of immature mouse (1.5 × 1.5 × 1.5 mm) or rabbit (2.0 × 2.0 × ≃3.0 mm) testis were cryopreserved, thawed and transplanted into mouse testes. Histological techniques were used to determine the presence of spermatogenesis, which was restored in both mouse and rabbit testicular pieces, and led to the production of mature sperm after both cryopreservation and syngeneic or xenogeneic transplantation into mouse testes. Using sperm developed in the frozen-thawed transplants, mouse offspring were born after in-vitro microinsemination. Furthermore, rabbit offspring were obtained using rabbit sperm that developed in fresh transplants in a xenogeneic surrogate mouse. Conclusions: This approach of 'testicular tissue banking' is a promising technique for the preservation of fertility in prepubertal male oncology patients. Xenogeneic transplantation into immunodeficient mice may provide a system for study spermatogenic failure in infertile men.

Scopus

Birth of offspring following transplantation of cryopreserved immature testicular pieces and in-vitro microinsemination

Shinohara, T; Inoue, K; Ogonuki, N; Kanatsu-Shinohara, M; Miki, H; Nakata, K; Kurome, M; Nagashima, H; Toyokuni, S; Kogishi, K; Honjo, T; Ogura, A

HUMAN REPRODUCTION Vol. 17 ( 12 ) page： 3039 - 3045 2002.12

A novel method of "preparative" myocardial protection using green tea polyphenol in oral uptake

Miwa, S; Saito, M; Yamazaki, K; Toyokuni, S; Hyon, SH; Fujita, M; Komeda, M

CIRCULATION Vol. 106 ( 19 ) page： 345 - 345 2002.11

Oxydative stress persists in nearby LV area after myocardial infarction (MI) and shows two peaks

Miwa, S; Toyokuni, S; Tanaka, S; Nishina, T; Nomoto, T; Yamazaki, K; Ueda, K; Nishimura, K

CIRCULATION Vol. 106 ( 19 ) page： 312 - 312 2002.11

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Papers - TOYOKUNI Shinya