著者

Hataitip TRISOMBOON Suchinda MALAIVIJITNOND Wichai CHERDSHEWASART Gen WATANABE Kazuyoshi TAYA

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

vol.52, no.4, pp.537-542, 2006 (Released:2006-08-25)

参考文献数

30

被引用文献数

17 19

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To investigate the estrogenic effect of Pueraria mirifica (PM), a Thai herbal plant that contains many phytoestrogens, sexual skin coloration was studied in cynomolgus monkeys. Aged menopausal monkeys were divided into three groups. Each group (n=3) was fed 10, 100, or 1,000 mg of PM daily. The treatment schedule was divided into three periods, a 30-day pre-treatment period, 90-day treatment period, and 60-day post-treatment period. The results show that the sexual skin exhibited reddish coloration within 24 h after PM-treatment and remained this way for the first half of the PM-feeding period. The changes in sexual skin coloration were not dose-dependent. The present results indicate that PM had estrogenic action by increasing reddish sexual skin coloration in aged menopausal monkeys.

著者

Hataitip TRISOMBOON Suchinda MALAIVIJITNOND Juri SUZUKI Yuzuru HAMADA Gen WATANABE Kazuyoshi TAYA

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

vol.50, no.6, pp.639-645, 2004 (Released:2005-01-07)

参考文献数

35

被引用文献数

18 19

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To determine the effect of Pueraria mirifica (PM) on serum parathyroid hormone (PTH) and calcium levels on aged menopausal monkeys (Macaca fascicularis), subjects were treated with 10, 100, or 1,000 mg/day of PM. Blood samples were collected every 5 days for 30, 90, and 60 days during pre-treatment, treatment, and post-treatment periods, respectively. Sera were assayed for PTH, estradiol, and calcium levels. PM-1,000 had the strongest effect on the decrease in PTH (0.001<P≤0.05) and calcium levels (0.001<P≤0.03) during the treatment period. PTH levels remained low for the first 15 days of the post-treatment period (0.01≤P ≤0.05). PM-10 induced a significant decrease in PTH level on day 80 (P=0.02) during the treatment period and a significant decrease in calcium level on day 75 (P<0.01). There were no changes in serum PTH and calcium levels throughout the study period in the PM-100 group. Estradiol levels decreased significantly during the treatment period in all treatment groups. The results suggest that long-term treatment with 1,000 mg/day of PM decreases serum PTH and calcium levels in aged menopausal monkeys, indicating that PM ameliorates bone loss caused by estrogen deficiency.

著者

Thanida SANANMUANG Nawapen PHUTIKANIT Catherine NGUYEN Sukanya MANEE-IN Mongkol TECHAKUMPHU Theerawat THARASANIT

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.2012-116, (Released:2013-01-25)

被引用文献数

4 6 1

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Developmental competence and quality of in vitro produced embryos has been demonstrated to be lower than in vivo derived embryos. This study aimed specifically to determine the effects of in vitro culture of feline embryos using various culture densities on developmental competence and expression of stress- and apoptotic-related genes in terms of heat shock protein 70 (HSP70) and apoptotic-related (BAX and BCL-2) gene expressions. In experiment 1, we characterized the inducible form of a feline-specific HSP70 mRNA sequence, as it has not been previously reported. The primers for feline HSP70 mRNA were synthesized and tested on heat-treated cat fibroblasts. In experiment 2, feline embryos were cultured at different culture densities (embryo:culture volume; 1:1.25, 1:5 and 1:20). The developmental competence was determined along with HSP70, BAX and BCL-2 transcript abundances using quantitative RT-PCR. In vivo derived embryos were used as a control group. A partial cat HSP70 mRNA sequence (190 bp) was characterized and exhibited high nucleotide identity (93 to 96%) with other species. Cleaved embryos cultured at high density (1:1.25) developed to blastocysts at a lower rate than those generated from lower densities. Irrespective of the culture densities used, in vitro cultured blastocysts showed increased levels of HSP70 and BAX transcripts compared with in vivo counterparts. Blastocysts derived from the highest culture density (1:1.25) showed higher levels of upregulation of HSP70 and BAX transcripts than those cultured at lower culture densities (1:5 and 1:20). In conclusion, increased levels of pro-apoptotic (BAX) and stress-response (HSP70) transcripts correlated with developmental incompetence of embryos cultured at high embryonic density, indicating that stress accumulated during in vitro embryo culture affected the fate for embryo development and quality.

著者

Narong TIPTANAVATTANA Chommanart THONGKITTIDILOK Mongkol TECHAKUMPHU Theerawat THARASANIT

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.2012-130, (Released:2013-01-25)

被引用文献数

7 15 3

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Spermatogonial stem cells (SSCs) function to regulate the balance of self-renewal and differentiation of male gametes. SSCs have been successfully isolated and cultured in vitro in several species, but not in feline. Therefore, in this study, we aimed to culture and characterize feline SSCs. In experiment 1, testes (n=5) from different pubertal domestic cats were cryosectioned and fluorescently immunolabeled to examine the expression of SSC (GFRα-1), differentiated spermatogonium (c-kit) and germ cell (DDX-4) markers. In experiments 2 and 3, testicular cells were digested and subsequently cultured in vitro. The resultant presumptive SSC colonies were then collected for SSC identification (experiment 2), or further cultured in vitro on feeder cells (experiment 3). Morphology, gene expression and immunofluorescence were used to identify the SSCs. Experiment 1 demonstrated that varying types of spermatogenic cells existed and expressed different germ cell/SSC makers. A rare population of putative SSCs located at the basement membrane of the seminiferous tubules was specifically identified by co-expression of GFRα-1 and DDX-4. Following enzymatic digestion, grape-like colonies formed by 13-15 days of culture. These colonies expressed GFRA1 and ZBTB16, but did not express KIT. Although we successfully isolated and cultured feline SSCs in vitro, the SSCs could only be maintained for 57 days. In conclusion, this study demonstrates, for the first time, that putative SSCs from testes of pubertal domestic cats can be isolated and cultured in vitro. These cells exhibited SSC morphology and expressed SSC-specific genes. However, long-term culture of these putative SSCs was compromised.

著者

Qing LI Yong FAN Xiaofang SUN Yanhong YU

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.2012-109, (Released:2012-11-09)

被引用文献数

2 13

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The ectopic expression of transcription factors for reprogramming human somatic cells to a pluripotent state represents a valuable resource for the development of in vitro-based models for human disease and has great potential in regenerative therapies. However, the majority of studies have used skin fibroblasts to generate induced pluripotent stem cells (iPSCs) that typically require the enforced expression of several transcription factors, thereby posing a mutagenesis risk by the insertion of viral transgenes. To reduce this risk, iPSCs have been generated with OCT4 and KLF4 from human neural stem cells that endogenously express the remaining reprogramming factors. However, human neural stem cells are rare and difficult to obtain. Here, we show that iPSCs can be generated from human amniotic fluid cells (hAFCs) with two transcription factors: OCT4 and KLF4. Furthermore, iPSCs can be readily derived from hAFCs in a feeder-free conditions, thereby eliminating the potential variability caused by using feeder cells. Our results indicate that hAFCs represent an accessible source of cells that can be reprogrammed into pluripotent stem cells with two Yamanaka factors. Therefore, hAFCs may become a preferred cell type in the future for safe reprogramming without any exogenous genetic material.

著者

Ayumi HASEGAWA Kazuya YONEZAWA Akihiko OHTA Keiji MOCHIDA Atsuo OGURA

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

vol.58, no.1, pp.156-161, 2012 (Released:2012-03-22)

参考文献数

30

被引用文献数

1 18 4

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The rapid increase in the number of genetically modified mouse strains has produced a high demand for their frozen spermatozoa from laboratories and mouse banking facilities. Historically, plastic straws have been used preferentially as containers for frozen mammalian spermatozoa because spermatozoa frozen in plastic straws have a high survival rate after thawing. However, plastic straws are more fragile and are used less often than the cryotubes used for conventional cell freezing. In this study, we sought to develop a new protocol for sperm freezing using cryotubes as the container to increase the accessibility of mouse sperm cryopreservation. Epididymal spermatozoa were collected from mature ICR or C57BL/6J (B6) males and were suspended in 18% raffinose and 3% skim milk solution. We then optimized the following conditions using the sperm survival rate as an index: 1) distance of cryotubes from the surface of the liquid nitrogen at freezing, 2) volume of the sperm suspension in the cryotube and 3) temperature of warming sperm during thawing. The best result was obtained when cryotubes containing 10 μl of sperm suspension were immersed 1 cm below the surface of the liquid nitrogen and then thawed at 50 C. The fertilization rates using spermatozoa frozen and thawed using this method were 63.1% in ICR mice and 28.2% in B6 mice. The latter rate was increased to 62.3% by adding reduced glutathione to the fertilization medium. After embryo transfer, 68% and 62% of the fertilized oocytes developed into normal offspring in the ICR and B6 strains, respectively. These results show that cryotubes can be used for cryopreservation of mouse spermatozoa under optimized conditions. This protocol is easy and reproducible, and it may be used in laboratories that do not specialize in sperm cryopreservation.

著者

Yong FAN Yumei LUO Xinjie CHEN Qing LI Xiaofang SUN

出版者

The Society for Reproduction and Development

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.1204060449, (Released:2012-04-13)

被引用文献数

29 36

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Induced pluripotent stem cells (iPSCs) derived from somatic cells of patients represent a powerful tool for biomedical research and may have a wide range of applications in cell and gene therapy. However, the safety issues and the low efficiency associated with generating human iPSCs have limited their usage in clinical settings. The cell type used to create iPSCs can significantly influence the reprogramming efficiency and kinetics. Here, we show that amniotic fluid cells from the prenatal diagnosis of a β-thalassemia patient can be efficiently reprogrammed using a doxycycline (DOX)-inducible humanized version of the single lentiviral “stem cell cassette” vector flanked by loxP sites, which can be excised with Cre recombinase. We also demonstrated that the patient-derived iPSCs can be characterized based on the expression of pluripotency markers, and they can be differentiated into various somatic cell types in vitro and in vivo. Moreover, microarray analysis demonstrates a high correlation coefficient between human β-thalassemia iPS cells and human embryonic stem (hES) cells but a low correlation coefficient between human β-thalassemia amniotic fluid cells and human β-thalassemia iPS cells. Our data suggest that amniotic fluid cells may be an ideal human somatic cell resource for rapid and efficient generation of patient-specific iPS cells.

著者

Noboru MANABE Akira MYOUMOTO Yoshihiro KIMURA Yuzuru IMAI Miki SUGIMOTO Hajime MIYAMOTO Kazuhiro SAKAMAKI Yoshinori OKAMURA Manabu FUKUMOTO

出版者

THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

vol.42, no.6, pp.j135-j141, 1996 (Released:2010-10-20)

被引用文献数

1 2

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哺乳類の卵母細胞は成熟する過程で選抜が行われて大部分の卵母細胞は選択的に死滅するが,この制御機構は未だ不明である.顕微鏡下に卵胞構成細胞を容易に分離調製できるブタ卵胞を材料として,二次卵胞から胞状卵胞に至る過程の卵胞退行における顆粒層細胞アポトーシスの役割とその制御について調べ,次のような結果を得た.はじめにTUNEL法および電顕法にて in situに卵胞退行にともなう顆粒層細胞アポトーシスの推移を観察した結果、内腔側の細胞から始まったアポトーシスは基底膜側に広がること,隣接する顆粒層細胞がアポトーシス小体を取り込むこと,および退行の進行した卵胞でも卵丘細胞にはアポトーシスが認められないことが分かった.なお,アポトーシスの開始は,ウシでは基底膜側から,齧歯類ではランダムであり,種属差があることも分かった.組織学的に見いだされた顆粒層細胞と卵丘細胞間の差異は生化学的に測定したエンドヌクレアーゼ活性の差異としても確認できた.すなわち,顆粒層細胞でのみ中性Ca2+/Mg2+-依存性エンドヌクレアーゼ活性の上昇を認めた.なお,ブタ顆粒層細胞アポトーシスには中性Ca2+-依存性,中性Mg2+-依存性および酸性カチオン非依存性エンドヌクレアーゼは関与しないことが分かった.ついで顆粒層細胞にアポトーシスを誘導できる単クローン抗体を作成し,これによって誘導される顆粒層細胞アポトーシスの進行がシステインプロテアーゼ阻害剤によって阻害されることをフローサイトメトリー法およびDNA電気泳動法にて確認した.顆粒層細胞では,アポトーシスシグナルの細胞内伝達にインターロイキン1β 転換酵素様プロテアーゼが関与していると考えられる.

著者

Hamayun KHAN Ken Takeshi KUSAKABE Shoichi WAKITANI Masato HIYAMA Ai TAKESHITA Yasuo KISO

出版者

日本繁殖生物学会

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

pp.1112200427-1112200427, (Released:2011-12-22)

被引用文献数

6 16

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Nitric oxide synthase (NOS) is a key regulator of angiogenesis and embryogenesis in the mammalian reproductive process. Here, we attempted to clarify the expression and localization of inducible and endothelial NOS (iNOS and eNOS) in the developing rabbit placenta. Real-time RT-PCR analysis indicated that iNOS mRNA was significantly upregulated during pregnancy, and then significantly decreased after d18 (completion of the placentation period). The eNOS mRNA was also enhanced in the pregnant uteri and gradually decreased near the term of pregnancy. Western blot analysis also showed elevation of the iNOS and eNOS protein levels during the course of pregnancy. Immunohistochemical study revealed distinct localizations of iNOS along the radial arteries and eNOS at the spiral arteries and arterial sinuses in the developing placenta. This may reflect that iNOS and eNOS participate in pregnancy success through placentation-specific vascular formation and by supporting adequate blood circulation in the rabbit placenta.

著者

Mutai Hideki Hattori Naka Kamei Takayuki AIKAWA Jun-ichi SHIOTA Kunio OGAWA Tomoya

出版者

THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

雑誌

The Journal of reproduction and development (ISSN:09168818)

巻号頁・発行日

vol.41, no.4, pp.353-359, 1995-11-01

被引用文献数

1

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One of the target tissues for prolactin (PRL) family members is the central nervous sytem (CNS) in adult animals. In this study, we tried to detect PRL receptor (PRL-R) mRNAs in fetal rat brains. mRNAs extracted from various tissues, including the brain, on embryonic days 12 (E12), 14, 16, 18 and 20 were subjected to reverse transcriptional polymerase chain reaction (RT-PCR) using two sets of primers specific to the short and long forms of the PRL-R. The short form cDNA band (330 bp) was detected in the fetal brain from E12 to 20, whereas the long band (420 bp) was not apparently observed, suggesting that the developing fetal brain is a target for PRL family members. Short form PRL-R mRNA was also expressed in the liver (E20), heart (E12 to 20), intestine (E16 to 20) and forelimb (E20), and long form PRL-R was observed in the heart on E20. Semi-quantitative PCR analysis revealed that short form PRL-R mRNA expression in the fetal brain was as biphasic: constant on E12 and 14, and increased progressively on E18 and 20. Thus, PRL-R gene expression is initiated very early in developing fetal tissues, including the CNS.

著者

Itagaki Yoshiaki Kimura Naoko Yamanaka Masaya MUNETA Yoshihiro

出版者

THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

雑誌

The Journal of reproduction and development (ISSN:09168818)

巻号頁・発行日

vol.43, no.1, pp.53-58, 1997-02-01

被引用文献数

2

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This study was undertaken to determine the relationship between the first cleavage division of <i>in vitro</i> produced (IVP) bovine embryos and the following few cleavage divisions in culture. The "early-", "intermediate-" and "late-" cleaving embryos presented at 22, 26 and 30 h post insemination (hpi) were cultured separately until they were fixed and stained with Hoechst 33342 at the end of each culture period (30, 48, 72, 96 and 120 hpi). Distributions of embryos that underwent the first cleavage at 22, 26 and 30 hpi were 21.5, 34.7 and 10.0%, respectively. No significant difference among the mean number of cells in the 3 groups was observed until 48 hpi. At 72 hpi, the number of cells in the early-cleaving embryos (10.8 ± 0.5) was higher than those in the intermediate- and late-cleaving embryos (8.7 ± 0.4 and 6.5 ± 0.6, respectively). Higher numbers of cells were also observed in the early-cleaving embryos (18.0 ± 1.5) at 120 hpi, compared with those in the intermediate- and late-cleaving embryos (12.5 ± 0.8 and 11.4 ± 1.5, respectively). More than 80% of the early- and intermediate-cleaving embryos had completed the second cleavage division at 48 hpi, whereas that of the late-cleaving embryos was lower (56.5%). No difference in the proportion of >8-cell embryos among the 3 groups was observed at 48 hpi (22.9 vs 24.7 vs 26.1%). However, the completion of the third cleavage in the early-cleaving embryos was higher than in the intermediate- and late-cleaving embryos at 72 hpi (65.4 vs 49.4 vs 25.0%), and from there onwards (96 hpi; 81.1 vs 63.2 vs 45.5%, 120 hpi; 83.6 vs 62.2 vs 65.0%). The proportion of >16-cell embryos in the early-cleaving embryos was also higher than that of the intermediate- and late-cleaving embryos between 96 hpi (18.9 vs 2.9 vs 0.0%) and 120 hpi (49.1 vs 22.0 vs 25.0%). Our results suggest that the development of IVP bovine embryos is possibly controlled as early as at the 2-cell stage, and the differences between the early and late cleaving embryos are associated with 1) the developmental delay or arrest of embryos during the second cleavage, and 2) the lengthening of the third cleavage, which seems to be related to the subsequent developmental ability of the embryos.

著者

Midori YOSHIDA Takasumi SHIMOMOTO Sayumi KATASHIMA Gen WATANABE Kazuyoshi TAYA Akihiko MAEKAWA

出版者

THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

雑誌

Journal of Reproduction and Development (ISSN:09168818)

巻号頁・発行日

vol.50, no.3, pp.349-360, 2004 (Released:2004-06-24)

参考文献数

56

被引用文献数

23 32

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Effects of maternal exposure to low doses of bisphenol A (BPA), including those comparable with human exposure levels, on growth and development of the female reproductive system and uterine carcinogenesis in Donryu rats were investigated. Dams were administered BPA (0, 0.006 and 6 mg/kg/day) daily by gavage from gestation day 2 up to the day before weaning (postnatal day 21 at offspring). The serum levels of BPA were significantly elevated in the dams receiving 6 mg/kg/day, however, BPA levels in the milk of dams, and those in the serum and liver of offspring were similar between control and treated groups. The treatment did not exert any influences on uterine development including weight, gland genesis and estrogen receptor α expression, vaginal opening and gonadotropin secretion in the female offspring up to puberty. After maturation, no effects were evident with regard to estrous cyclicity in female offspring treated with BPA. In addition, the treatment had no effects on age-related morphological changes of the reproductive and endocrine organs and uterine carcinogenesis until 15 months of age. The results demonstrate that maternal exposure to BPA at levels comparable to human exposure did not have any effects on the female reproductive system of offspring in rats. In addition, BPA was also found in the serum, milk and liver of control dams and pups, and low levels of BPA were detected in drinking water and pellet diet. The present study showed that the experimental animals were also exposed to environmental BPA in the animal room.