Faculty Profiles - TOYOKUNI Shinya

Papers - TOYOKUNI Shinya

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Adenovirus-mediated gene delivery into mouse spermatogonial stem cells Reviewed

Masanori Takehashi, Mito Kanatsu-Shinohara, Kimiko Inoue, Narumi Ogonuki, Hiromi Miki, Shinya Toyokuni, Atsuo Ogura, Takashi Shinohara

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Vol. 104 ( 8 ) page： 2596 - 2601 2007.2

Language：English Publishing type：Research paper (scientific journal) Publisher：NATL ACAD SCIENCES

Spermatogonial stem cells represent a self-renewing population of spermatogonia, and continuous division of these cells supports spermatogenesis throughout the life of adult male animals. Previous attempts to introduce adenovirus vectors into spermatogenic cells, including spermatogonial stem cells, have failed to yield evidence of infection, suggesting that male germ cells may be resistant to adenovirus infection. In this study we show the feasibility of transducing spermatogonial stem cells by adenovirus vectors. When testis cells from ROSA26 Cre reporter mice were incubated in vitro with a Cre-expressing adenovirus vector, Cre-mediated recombination occurred at an efficiency of 49-76%, and the infected spermatogonial stem cells could reinitiate spermatogenesis after transplantation into seminiferous tubules of infertile recipient testes. No evidence of germ-line integration of adenovirus vector could be found in offspring from infected stem cells that underwent Cre-mediated recombination, which suggests that the adenovirus vector infected the cells but did not stably integrate into the germ line. Nevertheless, these results suggest that adenovirus may inadvertently integrate into the patient's germ line and indicate that there is no barrier to adenovirus infection in spermatogonial stem cells.

DOI： 10.1073/pnas.0609282104

Adult pancreatic islets require differential pax6 gene dosage Reviewed

Akihiro Hamasaki, Yuichiro Yamada, Takeshi Kurose, Nobuhiro Ban, Kazuaki Nagashima, Akira Takahashi, Shimpei Fujimoto, Dai Shimono, Michio Fujiwara, Shinya Toyokuni, Yutaka Seino, Nobuya Inagaki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS Vol. 353 ( 1 ) page： 40 - 46 2007.2

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Pax6, a paired homeodomain transcription factor, plays crucial roles in morphogenesis of eye, central nervous system, and pancreatic islets. Recently, heterozygosity for pax6 mutation has been reported in some individuals with glucose intolerance and aniridia. To investigate the role of pax6 for pancreatic islet function, we examined the pancreatic phenotype of small eye rat strain (rSey(2)) with a point mutation in the pax6 locus resulting in truncated PAX6 proteins. Analyses of the insulin secretory profile of heterozygous rSey(2)/+ revealed that insulin secretion is significantly increased in response to membrane-depolarizing stimuli such as arginine, tolbutamide, and KCl. The processes of insulin granule exocytosis were suggested to be enhanced in rSey(2)/+. On the other hand, pancreatic insulin and glucagon content and islet architecture in rSey(2)/+ showed no significant differences compared to wild-type. These findings indicate differential requirements for pax6 gene dosage in displaying function and maintaining architecture of adult pancreatic islets. (c) 2006 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2006.11.105

Generation of insulin-secreting cells from pancreatic acinar cells of animal models of type 1 diabetes Reviewed

Masaaki Okuno, Kohtaro Minami, Akinori Okumachi, Kazumasa Miyawaki, Norihide Yokoi, Shinya Toyokuni, Susumu Seino

AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM Vol. 292 ( 1 ) page： E158 - E165 2007.1

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER PHYSIOLOGICAL SOC

We recently found that pancreatic acinar cells isolated from normal adult mouse can transdifferentiate into insulin-secreting cells in vitro. Using two different animal models of type 1 diabetes, we show here that insulin- secreting cells can also be generated from pancreatic acinar cells of rodents in the diabetic state with absolute insulin deficiency. When pancreatic acinar cells of streptozotocin-treated mice were cultured in suspension in the presence of epidermal growth factor and nicotinamide under low-serum condition, expressions of insulin genes gradually increased. In addition, expressions of other pancreatic hormones, including glucagon, somatostatin, and pancreatic polypeptide, were also induced. Analysis by the Cre/loxP-based direct cell lineage tracing system revealed that these newly made cells originated from amylase-expressing pancreatic acinar cells. Insulin secretion from the newly made cells was significantly stimulated by high glucose and other secretagogues. In addition, insulin- secreting cells were generated from pancreatic acinar cells of Komeda diabetes-prone rats, another animal model of type 1 diabetes. The present study demonstrates that insulin- secreting cells can be generated by transdifferentiation from pancreatic acinar cells of rodents in the diabetic state and further suggests that pancreatic acinar cells represent a potential source of autologous transplantable insulin- secreting cells for treatment of type 1 diabetes.

DOI： 10.1152/ajpendo.00180.2006

Leukemia inhibitory factor enhances formation of germ cell colonies in neonatal mouse testis culture

Mito Kanatsu-Shinohara, Kimiko Inoue, Narumi Ogonuki, Hiromi Miki, Shosei Yoshida, Shinya Toyokuni, Jiyoung Lee, Atsuo Ogura, Takashi Shinohara

BIOLOGY OF REPRODUCTION Vol. 76 ( 1 ) page： 55 - 62 2007.1

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

Spermatogonial stem cells continuously divide in the testis to support spermatogenesis throughout the life of adult male animals. Although very few spermatogonial stem cells are present in vivo, we recently succeeded in expanding these cells in vitro. Germ cells from postnatal testes were able to proliferate in the presence of several types of cytokines, and they formed uniquely shaped colonies of spermatogonia (germline stem or GS cells). These cells reinitiated normal spermatogenesis when transplanted into seminiferous tubules. However, much remains unknown about the contributions of cytokines to successful stem cell culture. In the present study, we examined the role of leukemia inhibitory factor (LIF) in GS cell culture. We found that the addition of LIF to newborn testis cell culture enhances the formation of germ cell colonies. Ciliary neurotrophic factor, but not oncostatin M, had the same effect, although they both bind to the IL-6ST (gp130) receptor. On the other hand, GS cells could be established from pup or adult testes in the absence of LIF. No phenotypic or functional difference was found between GS cells established from different stages, and normal offspring were born from pup-derived GS cells that had been maintained in the absence of LIF, indicating that LIF per se is not involved in the self-renewal of GS cells. These results demonstrate that LIF is useful in the initiation of GS cell culture and suggest that LIF or a related cytokine is involved in the maturation of gonocytes into spermatogonia.

DOI： 10.1095/biolreprod.106.055863

Akt mediates self-renewal division of mouse spermatogonial stem cells. Reviewed

Lee J, Kanatsu-Shinohara M, Inoue K, Ogonuki N, Miki H, Toyokuni S, Kimura T, Nakano T, Ogura A and Shinohara T.

Development Vol. 134 page： 1853-1859 2007

Association of microRNA-34a overexpression with proliferation is cell type-dependent. Reviewed

Dutta KK, Zhong Y, Liu YT, Yamada T, Akatsuka S, Hu Q, Yoshihara M, Ohara H, Takehashi M, Shinohara T, Masutani H, Onuki J and Toyokuni S.

Cancer Sci. Vol. 98 page： 1845-1852 2007

FasL expression in hepatic antigen-presenting cells and phagocytosis of apoptotic T cells by FasL+ Kupffer cells are indicators of rejection activity in human liver allografts. Reviewed

Miyagawa-Hayashino A, Tsuruyama T, Egawa H, Haga H, Sakashita H, Okuno T, Toyokuni S, Tamaki K, Yamabe H, Manabe T and Uemoto S.

Am J. Pathol. Vol. 171 page： 1499-1508 2007

Production of knockout mice by gene targeting in multipotent germline stem cells. Reviewed

Takehashi M, Kanatsu-Shinohara M, Miki H, Lee J, Kazuki Y, Inoue K, Ogonuki N, Toyokuni S, Oshimura M, Ogura A and Shinohara T.

Dev. Biol. Vol. 312 page： 344-352 2007

Visualization and characterization of UVB-induced reactive oxygen species in a human skin equivalent model. Reviewed

Hakozaki T, Date A, Yoshii T, Toyokuni S, Yasui H and Sakurai H.

J. Dermatol. Res. Vol. 300 (Supplement 1) page： S51-56 2007

*Chronic oxidative stress causes amplification and overexpresson of ptprz1 protein tyrosine phosphatase to activate beta-catenin pathway. Reviewed

Liu YT, Shang DH, Akatsuka S, Ohara H, Dutta KK, Mizushima K, Naito Y, Yoshikawa T, Izumiya M, Abe K, Nakagama H, Noguchi N and Toyokuni S.

Am. J. Pathol. Vol. 171 page： 1978-1988 2007

ARMET is a soluble ER protein induced by the unfolded protein response via ERSE-II element. Reviewed

Mizobuchi N, Hoseki J, Kubota H, Toyokuni S, Nozaki J, Naitoh M, Koizumi A and Nagata K.

Cell Struct. Funct. Vol. 32 page： 41-50 2007

Generation of insulin-secreting cells from pancreatic acinar cells of animal models of type 1 diabetes. Reviewed

Okuno M, Minami K, Okumachi A, Miyawaki K, Yokoi N, Toyokuni S, Seino S.

Am. J. Physiol. Endocrinol. Metab. Vol. 292 page： E158-165 2007

Oxidative stress is related to formation of Antoni B pattern and eosinophilic hyaline droplets in schwannoma. Reviewed

Yokoo H, Oishi T, Isoda, K, Nakazato Y and Toyokuni S.

Neuropathology Vol. 27 page： 237-244 2007

Angiotensin II, oxidative stress, and extracellular matrix degradation during transition to LV failure in rats with hypertension. Reviewed

Takenaka H, Kihara Y, Iwanaga Y, Onozawa Y, Toyokuni S, Kita T.

J. Mol. Cell. Cardiol. Vol. 41 page： 989-997 2007

Leukemia inhibitory factor enhances formation of germ cell colonies in neonatal mouse testis culture.

Kanatsu-Shinohara M, Inoue K, Ogonuki N, Miki H, Yoshida S, Toyokuni S, Lee J, Ogura A and Shinohara T.

Biol. Reprod. Vol. 76 page： 55-62 2007

Adult pancreatic islets require differential pax6 gene dosage. Reviewed

Hamasaki A, Yamada Y, Kurose T, Ban N, Nagashima K, Takahashi A, Fujimoto S, Shimono D, Fujiwara M, Toyokuni S, Seino Y and Inagaki N.

Biochem. Biophys. Res. Commun. Vol. 353 page： 40-46 2007

Adenovirus-mediated gene delivery into mouse spermatogonial stem cells. Reviewed

Takehashi M, Kanatsu-Shinohara M, Inoue K, Ogonuki N, Miki H, Toyokuni S, Ogura A, and Shinohara T.

Proc. Natl. Acad. Sci. USA Vol. 104 page： 2596-2601 2007

Pathological investigation of oxidative stress in the post-genomic era. Invited Reviewed

Toyokuni S and Akatsuka S.

Pathol. Int. Vol. 57 page： 461-473 2007

Overexpressin of a transcription factor LYL-1 induces T- and B-cell lymphoma in mice. Reviewed

Zhong Y, Li J, Hiai H, Toyokuni S* and Yamada Y* (*corresponding author).

Oncogene Vol. 26 page： 6937-6947 2007

Analysis of genes associated with karyo-megalic cells in rat kidney after ferric nitilotriacetate treatment International journal

Lee, WH; Toyokuni, S

FREE RADICAL BIOLOGY AND MEDICINE Vol. 43 page： S24 - S24 2007

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