Papers - KUWAYAMA Takehito
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Severe maternal stress alters placental function, resulting in adipose tissue and liver dysfunction in offspring of mice Reviewed
Kondo R, Ozawa R, Satomi T, Funabayashi K, Iwata H, Kuwayama T, Shirasuna K.
Mol Cell Endocrinol 560 111814 2023
Language：English Publishing type：Research paper (scientific journal)
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PCR 法によるホロホロチョウ（Numida meleagris）の性判別 Reviewed
南村昌孝・石田和真・黒澤 亮・高橋幸水・白砂孔明・岩田尚孝・小川 博・桑山岳人
日本家禽学会誌 59 ( J1 ) J1 - J5 2022.04
Authorship：Corresponding author Language：Japanese Publishing type：Research paper (scientific journal) Publisher：日本家禽学会
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Acetoacetate is a trigger of NLRP3 inflammasome activation in bovine peripheral blood mononuclear cells Reviewed
Onizawa Y, Katoh T, Miura R, Konda K, Noguchi T, Iwata H, Kuwayama T, Hamano S, Shirasuna K.
Vet Immunol Immunopathol 244 110370 2022
Language：English
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NLRP3 inflammasome is involved in testicular inflammation induced by lipopolysaccharide in mice Reviewed International journal
Sano M, Komiyama H, Shinoda R, Ozawa R, Watanabe H, Karasawa T, Takahashi M, Torii Y, Iwata H, Kuwayama T, Shirasuna K.
Am J Reprod Immunol 87 e13527 2022
Language：English Publishing type：Research paper (scientific journal)
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HIKATA Kiho, MOROHASHI Nanaho, TARU Maho, SUZUKI Yusuke, NAKAMURA Tomoaki, TAKEDA Masahiro, KUWAYAMA Takehito, SHIRASUNA Koumei
Japanese Journal of Zoo and Wildlife Medicine 26 ( 4 ) 127 - 134 2021.12
Language：Japanese Publisher：Japanese Society of Zoo and Wildlife Medicine
<p>The southern tamandua (<i>Tamandua tetradactyla</i>), classified in the genus Tamandua, belongs to the clade Xenarthra, which includes other species like sloths and anteaters. The blood hormone during the gestation period for one individual of southern tamandua was determined; however, there have been no reports on the hormonal changes during multiple pregnancy period. Therefore, long-term blood sampling of the same female individual was performed, and plasma progesterone (P4) or estradiol (E2) during a total of 6 gestation periods was determined in the same male-female pair. From the results of plasma P4 concentration during all 6 gestation periods, the gestation period was estimated as 156.8 ± 1.7 days (152-164 days) long. The P4 concentration was 0.6 ± 0.1 ng/ml before the pregnancy period, 13.2 ± 1.8 ng/ml during early pregnancy, 28.1 ± 4.3 ng/ml during mid pregnancy, and 48.2 ± 11.8 ng/ml in late pregnancy. Plasma E2 concentration gradually increased from 100 days before the delivery until the day of delivery. The estrous cycle was estimated to be 45.5 ± 2.4 days (37-52 days) long. Thus, we demonstrated that the blood steroid hormone kinetics during multiple pregnancies of the same pair of southern tamanduas. The increase in P4 concentration was observed prior to the increase in E2 concentration during the early stage of the pregnancy, suggesting that early pregnancy determination is possible by detecting a continuous increase in P4 concentration in the southern tamandua.</p>
DOI： 10.5686/jjzwm.26.127
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Mitochondrial reactive oxygen species regulate mitochondrial biogenesis in porcine embryos
KAGEYAMA Mio, ITO Jun, SHIRASUNA Koumei, KUWAYAMA Takehito, IWATA Hisataka
Journal of Reproduction and Development 67 ( 2 ) 141 - 147 2021
Language：English Publisher：The Society for Reproduction and Development
<p> The number of mitochondria in blastocysts is a potential marker of embryo quality. However, the molecular mechanisms governing the mitochondrial number in embryos are unclear. This study was conducted to investigate the effect of reduced mitochondrial reactive oxygen species (ROS) levels on mitochondrial biogenesis in porcine embryos. Oocytes were collected from gilt ovaries and activated to generate over 4 cell-stage embryos at day 2 after activation. These embryos were cultured in media containing either 0.1 μM MitoTEMPOL (MitoT), 0.5 μM Mitoquinol (MitoQ), or vehicle (ethanol) for 5 days to determine the rate of development to the blastocyst stage. The mitochondrial number in blastocysts was evaluated by real-time polymerase chain reaction (PCR). Five days after activation, the embryos (early morula stage) were subjected to immunostaining to determine the expression levels of NRF2 in the nucleus. In addition, the expression levels of <i>PGC1α</i> and <i>TFAM</i> in the embryos were examined by reverse transcription PCR. One day of incubation with the antioxidants reduced the ROS content in the embryos but did not affect the rate of development to the blastocyst stage. Blastocysts developed in medium containing MitoT had lower mitochondrial DNA copy numbers and ATP content, whereas MitoQ showed similar but insignificantly trends. Treatment of embryos with either MitoT or MitoQ decreased the expression levels of NRF2 in the nucleus and levels of <i>PGC1α</i> and <i>TFAM</i>. These findings indicate that reductions in mitochondrial ROS levels are associated with low mitochondrial biogenesis in embryos.</p>
DOI： 10.1262/jrd.2020-111
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HIRATA Yoshiki, KATSUKURA Yusuke, HENMI Yuka, OZAWA Ren, SHIMAZAKI Sayaka, KUROSAWA Akira, TORII Yasushi, TAKAHASHI Hironori, IWATA Hisataka, KUWAYAMA Takehito, SHIRASUNA Koumei
Journal of Reproduction and Development 67 ( 4 ) 257 - 264 2021
Language：English Publisher：The Society for Reproduction and Development
<p> Advanced maternal age is a risk factor for female infertility, and placental dysfunction is considered one of the causes of pregnancy complications. We investigated the effects of advanced maternal aging on pregnancy outcomes and placental senescence. Female pregnant mice were separated into three groups: young (3 months old), middle (8–9 months old), and aged (11–13 months old). Although the body weights of young and middle dams gradually increased during pregnancy, the body weight of aged dams only increased slightly. The placental weight and resorption rate were significantly higher, and live fetal weights were reduced in a maternal age-dependent manner. Although mRNA expression of senescence regulatory factors (p16 and p21) increased in the spleen of aged dams, mRNA expression of p16 did not change and that of p21 was reduced in the placenta of aged dams. Using a cytokine array of proteins extracted from placental tissues, the expression of various types of senescence-associated secretory phenotype (SASP) factors was decreased in aged dams compared with young and middle dams. The aged maternal placenta showed reduced immune cell accumulation compared with the young placenta. Our present results suggest that models using pregnant mice older than 8 months are more suitable for verifying older human pregnancies. These findings suggest that general cellular senescence programs may not be included in the placenta and that placental functions, including SASP production and immune cell accumulation, gradually decrease in a maternal age-dependent manner, resulting in a higher rate of pregnancy complications.</p>
DOI： 10.1262/jrd.2021-034
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Uncaria tomentosa extract (AC-11) improves pregnancy hypertension together with suppression of sFlt-1 and sEng Reviewed
26 127 - 132 2021
Language：English Publishing type：Research paper (scientific journal)
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β-hydroxybutyrate suppresses NLRP3 inflammasome-mediated placental inflammation and lipopolysaccharide-induced fetal absorption Reviewed International journal
Hirata Y, Shimazaki S, Suzuki S, Henmi Y, Komiyama H, Kuwayama T, Iwata H, Karasawa T, Takahashi M, Takahashi H, Shirasuna K.
J Reprod Immunol 148 103433 2021
Language：English Publishing type：Research paper (scientific journal)
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Mitochondrial reactive oxygen species regulate mitochondrial biogenesis in porcine embryos Reviewed
Kageyama M, Ito J, Shirasuna K, Kuwayama T, Iwata H.
J Reprod Dev 67 141 - 147 2021
Language：English Publishing type：Research paper (scientific journal)
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IL1B triggers inflammatory cytokine production in bovine oviduct epithelial cells and induces neutrophil accumulation via CCL2 Reviewed
Nakamura Y, Aihara R, Iwata H, Kuwayama T, Shirasuna K.
Am J Reprod Immunol 85 e13365 2021
Language：English
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MOROHASHI Nanaho, SANO Michiya, KATO Kosuke, TOYODA Hideto, NAKAMURA Tomoaki, TAKEDA Masahiro, KUWAYAMA Takehito, SHIRASUNA Koumei
Japanese Journal of Zoo and Wildlife Medicine 25 ( 4 ) 119 - 127 2020.12
Language：Japanese Publisher：Japanese Society of Zoo and Wildlife Medicine
<p> Capybara (<i>Hydrochoerus hydrochaeris</i>) is the biggest rodent in the world which lives in South America and well-known at Japanese zoos. Because the placental structure of capybara is similar to human placenta, the use of capybara as an experimental animal is also being considered. It is essential to understand the reproductive physiology of capybara for the planned breeding. However, there are few information about the reproductive physiology of capybara. In the present study, we tried to establish the method of blood sample collection and vaginal smear inspection using husbandary training (no anesthesia and retention). Then, the steroid hormone concentration in plasma and feces was examined to identify the estrous cycle and pregnancy in capybara. Plasma or fecal samples were collected from non-pregnant and pregnant capybaras and steroid hormones were determined by ELISA. In non-pregnant individuals, changes in blood progesterone (P<sub>4</sub>) and estrone-3-sulfate (E<sub>1</sub>S) concentrations showed clear estrous cycle-like changes for 3 cycles (average 11.7 ± 0.9 days). Change in blood E<sub>1</sub>S concentration were consistent with change in fecal estradiol (E<sub>2</sub>) and appearance of anucleated keratinized epithelial cells in vaginal smear inspection. Fecal E<sub>2</sub> concentrations of non-pregnancy capybara also changed like the estrous cycle. In fecal samples of pregnant capybara, the concentration of P<sub>4</sub> were higher than that in the non-pregnant capybara. Fecal P<sub>4</sub> and E<sub>2</sub> concentrations were higher during pregnancy and gradually reduced after parturition. These results suggest that the estrous cycle in capybara may be identified by determining plasma P<sub>4</sub>, E<sub>1</sub>S and fecal E<sub>2</sub> concentrations and vaginal smear inspection, and pregnancy of capybara can be predicted by measuring the P<sub>4</sub> concentration in feces.</p>
DOI： 10.5686/jjzwm.25.119
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SANO Michiya, SHIMAZAKI Sayaka, KANEKO Yasuaki, KARASAWA Tadayoshi, TAKAHASHI Masafumi, OHKUCHI Akihide, TAKAHASHI Hironori, KUROSAWA Akira, TORII Yasushi, IWATA Hisataka, KUWAYAMA Takehito, SHIRASUNA Koumei
Journal of Reproduction and Development 66 ( 3 ) 241 - 248 2020
Language：English Publisher：The Society for Reproduction and Development
<p> Maternal obesity is one of the major risk factors for pregnancy complications and is associated with low-grade chronic systemic inflammation due to higher levels of pro-inflammatory cytokines such as interleukin (IL)-1β. Pregnant women with obesity have abnormal lipid profiles, characterized by higher levels of free fatty acids, especially palmitic acid (PA). Previously, we reported that PA stimulated IL-1β secretion via activation of NLRP3 inflammasome in human placental cells. These observations led us to hypothesize that higher levels of PA induce NLRP3 inflammasome activation and placental inflammation, resulting in pregnancy complications. However, the effects of PA on NLRP3 inflammasome during pregnancy <i>in vivo</i> remain unclear. Therefore, PA solutions were administered intravenously into pregnant mice on day 12 of gestation. Maternal body weight was significantly decreased and absorption rates were significantly higher in PA-injected mice. The administration of PA significantly increased IL-1β protein and the mRNA expression of NLRP3 inflammasome components (<i>NLRP3</i>, <i>ASC</i>, and caspase-1) within the placenta. In murine placental cell culture, PA significantly stimulated IL-1β secretion, and this secretion was suppressed by a specific NLRP3 inhibitor (MCC950). Simultaneously, the number of macrophages/monocytes and neutrophils, together with the mRNA expression of these chemokines increased significantly in the placentas of PA-treated mice. Treatment with PA induced ASC assembling and IL-1β secretion in macrophages, and this PA-induced IL-1β secretion was significantly suppressed in NLRP3-knockdown macrophages. These results indicate that transient higher levels of PA exposure in pregnant mice activates NLRP3 inflammasome and induces placental inflammation, resulting in the incidence of absorption.</p>
DOI： 10.1262/jrd.2020-007
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KOBAYASHI Mitsuru, ITO Jun, SHIRASUNA Koumei, KUWAYAMA Takehito, IWATA Hisataka
Journal of Reproduction and Development 66 ( 6 ) 539 - 546 2020
Language：English Publisher：The Society for Reproduction and Development
<p> We examined the effect of ploidy on mitochondrial DNA (mtDNA) copy number in embryos and the amount of cell-free mitochondrial and nucleic DNA content (cf-mtDNA and cf-nDNA) in spent culture medium (SCM). Oocytes collected from the ovaries were matured, activated, incubated in medium containing cycloheximide (CHX) or CHX and cytochalasin B (CB) for 4.5 h to produce haploid or diploid embryos (H-group and D-group embryos). These embryos were cultured for 7 days, and the blastocysts and SCM were examined. The amount of mtDNA and nDNA was determined by real-time PCR. The rate of development to the blastocyst stage was higher for the D-group than for the H-group. Moreover, D-group blastocysts had less mtDNA compared to the H-group blastocysts. After activation, the mitochondrial content was constant before the blastocyst stage in D-group embryos, but increased earlier in H-group embryos. The amount of cf-mtDNA in the SCM of D-group blastocysts was greater than that of H-group blastocysts. However, when the cf-mtDNA in the SCM of 2 cell-stage embryos (day 2 post-activation) was examined, the amount of cf-mtDNA was greater in the H-group than in the D-group embryos. When D-group embryos were cultured for 7 days, a significant correlation was observed between the total cell number of blastocysts and cf-nDNA content in the SCM. Hence, although careful consideration is needed regarding the time point for evaluating mtDNA content in the embryos and SCM, this study demonstrates that mtDNA in the embryos and SCM was affected by the ploidy of the embryos.</p>
DOI： 10.1262/jrd.2020-097
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Interferon tau regulates cytokine production and cellular function in human trophoblast cell line. Reviewed
Tanikawa N, Seno K, Kawahara-Miki R, Kimura K, Matsuyama S, Iwata H, Kuwayama T, Shirasuna K
J Interferon & Cytokine Research 37 456 - 466 2017.10
Language：English Publishing type：Research paper (scientific journal)
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Advanced glycation end products and lipopolysaccharide stimulate interleukin-6 secretion via RAGE/TLR4-NF-kB-ROS pathways and resveratrol attenuates these inflammatory responses in mouse macrophages.
Ohtsu A, Shibutani Y, Seno K, Iwata H, Kuwayama T, Shirasuna K.
Experimental and Therapeutic Medicine 14 4364 - 4370 2017.08
Language：English Publishing type：Research paper (scientific journal)
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Age-associated mRNA expression changes in bovine endometrial cells in vitro. Reviewed
Tanikawa N, Ohtsu A, Kawahara-Miki R, Kimura K, Matsuyama S, Iwata H, Kuwayama T, Shirasuna K
Reprod Biol Endocrinol 63 2017.08
Language：English Publishing type：Research paper (scientific journal)
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Advanced glycation end products regulate interleukin-1β production in human placenta. Reviewed
Seno K, Sase S, Ozeki A, Takahashi H, Ohkuchi A, Suzuki H, Matsubara S, Iwata H, Kuwayama T, Shirasuna K
J Reprod Dev 63 401 - 408 2017.05
Language：English Publishing type：Research paper (scientific journal)
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Palmitic acid stimulates interleukin-8 via the TLR4/NF-kB/ROS pathway and induces mitochondrial dysfunction in bovine oviduct epithelial cells. Reviewed
Ohtsu A, Tanaka H, Seno K, Iwata H, Kuwayama T, Shirasuna K.
Am J Reprod Immunol 77 2017.02
Language：English Publishing type：Research paper (scientific journal)
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Promotion of glucose utilization by insulin enhances granulosa cell proliferation and developmental competence of porcine oocyte growth in vitro. Reviewed
Itami N, Munakata Y, Shirasuna K, Kuwayama T, Iwata H
Zygote 25 65 - 74 2016.12
Language：English Publishing type：Research paper (scientific journal)