著者

Adeleh ZABIHI Hamed Karami SHABANKAREH Hadi HAJARIAN Saheb FOROUTANIFAR

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.2018-102, (Released:2019-01-18)

被引用文献数

2

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This article released online on January 18, 2019 as advance publication was withdrawn from consideration for publication in The Journal of Reproduction and Development at author’s request.

著者

Blanca ALGARRA Verónica MAILLO Manuel AVILÉS Alfonso GUTIÉRREZ-ADÁN Dimitrios RIZOS María JIMÉNEZ-MOVILLA

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.64, no.5, pp.433-443, 2018 (Released:2018-10-12)

参考文献数

64

被引用文献数

17

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Previously, our group demonstrated that recombinant porcine oviductin (pOVGP1) binds to the zona pellucida (ZP) of in vitro-matured (IVM) porcine oocytes with a positive effect on in vitro fertilization (IVF). The fact that pOVGP1 was detected inside IVM oocytes suggested that this protein had a biological role during embryo development. The aim of this study was to evaluate the effects of pOVGP1 on bovine in vitro embryo development. We applied 10 or 50 µg/ml of pOVGP1 during IVF, embryonic in vitro culture (IVC), or both, to evaluate cleavage and embryo development. Blastocyst quality was assessed by analyzing the expression of important developmental genes and the survival rates after vitrification/warming. pOVGP1 was detected in the ZP, perivitelline space, and plasma membrane of blastocysts. No significant differences (P > 0.05) were found in cleavage or blastocyst yield when 10 or 50 µg/ml of pOVGP1 was used during IVF or IVC. However, when 50 µg/ml pOVGP1 was used during IVF + IVC, the number of blastocysts obtained was half that obtained with the control and 10 µg/ml pOVGP1 groups. The survival rates after vitrification/warming of expanded blastocysts cultured with pOVGP1 showed no significant differences between groups (P > 0.05). The use of pOVGP1 during IVF, IVC, or both, increased the relative abundance of mRNA of DSC2, ATF4, AQP3, and DNMT3A, the marker-genes of embryo quality. In conclusion, the use of pOVGP1 during bovine embryo in vitro culture does not affect embryo developmental rates but produces embryos of better quality in terms of the relative abundance of specific genes.

著者

Fuminori TANIHARA Maki HIRATA Nhien Thi NGUYEN Quynh Anh LE Takayuki HIRANO Takeshige OTOI

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.2018-116, (Released:2019-02-05)

被引用文献数

32

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Cytoplasmic microinjection (CI) of the CRISPR/Cas9 system enabled the induction of site-specific mutations in porcine zygotes and resulting pigs. However, mosaicism is a serious problem for genetically modified pigs. In the present study, we investigated suitable timing and concentration of CRISPR/Cas9 components for introduction into oocytes/zygotes by CI, to reduce mosaicism in the resulting blastocysts. First, we introduced 20 ng/μl of Cas9 protein and guide RNA (gRNA), targeting the α-1,3-galactosyltransferase (GalT) gene in oocytes before in vitro fertilization (IVF), in zygotes after IVF, or in oocytes/zygotes before and after IVF, twice. CI treatment had no detrimental effects on blastocyst formation rates. The highest value of the rate of mutant blastocysts was observed in zygotes injected after IVF. Next, we injected Cas9 protein and gRNA into zygotes after IVF at a concentration of 20 ng/μl each (20 ng/μl group) or 100 ng/μl each (100 ng/μl group). The ratio of the number of blastocysts that carried mutations to the total number of blastocysts examined in the 100 ng/μl group was significantly higher (P < 0.05) than that in the 20 ng/μl group. Although no blastocysts from the 20 ng/μl group carried a biallelic mutation, 16.7% of blastocysts from the 100 ng/μl group carried a biallelic mutation. In conclusion, increasing the concentration of Cas9 protein and gRNA is effective in generating biallelic mutant blastocysts. To reduce mosaicism, however, further optimization of the timing of CI, and the concentration of CRISPR/Cas9 components, is needed.

著者

Yoshihisa UENOYAMA Naoko INOUE Kei-ichiro MAEDA Hiroko TSUKAMURA

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.2018-110, (Released:2018-10-08)

被引用文献数

32

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Kisspeptin, identified as a natural ligand of GPR54 in 2001, is now considered as a master regulator of puberty and subsequent reproductive functions in mammals. Our previous studies using Kiss1 knockout (KO) rats clearly demonstrated the indispensable role of kisspeptin in gonadotropin-releasing hormone (GnRH)/gonadotropin secretion. In addition, behavioral analyses of Kiss1 KO rats revealed an organizational effect of kisspeptin on neural circuits controlling sexual behaviors. Our studies using transgenic mice carrying a region-specific Kiss1 enhancer-driven reporter gene provided a clue as to the mechanism by which estrogen regulates Kiss1 expression in hypothalamic kisspeptin neurons. Analyses of Kiss1 expression and gonadotropin secretion during the pubertal transition shed light on the mechanism triggering GnRH/gonadotropin secretion at the onset of puberty in rats. Here, we summarize data obtained from the aforementioned studies and revisit the physiological roles of kisspeptin in the mechanism underlying reproductive functions in mammals.

著者

KADOKAWA Hiroya

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.53, no.1, pp.121-125, 2007

被引用文献数

11

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Stress due to summer heat has adverse effects on reproduction in Holstein dairy cattle. Summer suppression of reproduction of Holsteins can pose an important economic problem, even in Hokkaido prefecture located in the northern region of Japan. Hokkaido is one of the most important dairy farming areas of Japan. This study is an attempt to clarify the seasonal differences in the parameters of luteinizing hormone (LH) response to exogenous gonadotropin releasing hormone (GnRH) in Sapporo, Hokkaido, Japan. A total of 12 prepubertal heifers received an injection with GnRH analogue intramuscularly in either May (n=4, May group), July (n=4, July group), or November (n=4, November group), and serial blood samples were collected to analyze the parameters of the LH response curve after GnRH injection. The parameters were as follows: the basal LH concentration, peak LH concentration, duration from the time of GnRH injection to the time of the peak LH concentration, and area under the LH response curve (AUC). There were no significant differences in the basal and peak LH concentrations or the AUC among the three groups. The July group reached the LH peak significantly (P<0.05) faster than the May group, but there was no significant difference with the November group. Therefore, the results of the present study do not demonstrate an effect of summer heat on the LH response to the exogenous GnRH in Holstein heifers.<br>

著者

Blanca ALGARRA Verónica MAILLO Manuel AVILÉS Alfonso GUTIÉRREZ-ADÁN Dimitrios RIZOS Maria JIMENEZ-MOVILLA

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.2018-058, (Released:2018-08-06)

被引用文献数

17

---

Previously, our group demonstrated that recombinant porcine oviductin (pOVGP1) binds to the zona pellucida (ZP) of in vitro-matured (IVM) porcine oocytes with a positive effect on in vitro fertilization (IVF). The fact that pOVGP1 was detected inside IVM oocytes suggested that this protein had a biological role during embryo development. The aim of this study was to evaluate the effects of pOVGP1 on bovine in vitro embryo development. We applied 10 or 50 µg/ml of pOVGP1 during IVF, embryonic in vitro culture (IVC), or both, to evaluate cleavage and embryo development. Blastocyst quality was assessed by analyzing the expression of important developmental genes and the survival rates after vitrification/warming. pOVGP1 was detected in the ZP, perivitelline space, and plasma membrane of blastocysts. No significant differences (P > 0.05) were found in cleavage or blastocyst yield when 10 or 50 µg/ml of pOVGP1 was used during IVF or IVC. However, when 50 µg/ml pOVGP1 was used during IVF + IVC, the number of blastocysts obtained was half that obtained with the control and 10 µg/ml pOVGP1 groups. The survival rates after vitrification/warming of expanded blastocysts cultured with pOVGP1 showed no significant differences between groups (P > 0.05). The use of pOVGP1 during IVF, IVC, or both, increased the relative abundance of mRNA of DSC2, ATF4, AQP3, and DNMT3A, the marker-genes of embryo quality. In conclusion, the use of pOVGP1 during bovine embryo in vitro culture does not affect embryo developmental rates but produces embryos of better quality in terms of the relative abundance of specific genes.

著者

梅津 元昭

出版者

THE SOCIETY FOR REPRODUCTION AND DEVELOPMENT

雑誌

家畜繁殖研究會誌 (ISSN:04530551)

巻号頁・発行日

vol.19, no.3, pp.87-93, 1973

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PMS注射による幼若雌ラットの排卵誘起に対し,アンチエストロジェン作用を有するクロミヘンを処理することにより排卵の抑制がみられるが,今回はその排卵抑制に対するエストロジェンおにびプロゲスチソの影響について種々の検討を加え,次の結果を得た。<BR>1) 24~26日令のラットを用い,3IUのPMSを注射後6時間にクロミヘン2.5mg/100 g b.w.を注射した時,排卵の抑制がみられ,排卵は1日遅れた。子宮除去ラットでも同様の結果が得られた。<BR>2) 上記のPMSおよびクロミヘン処理で,PMS注射後エストラジオールを種々の時間に単一皮下注射したところ, PMS注射後3時間のエストラジオール0.5μg, 5μg処理,6時間,9時間および51時間のそれぞれ5μg処理で高い排卵率が得られ,排卵抑制効果はほぼ打ち消された。<BR>3) 同様に, PMS処理後プロゲステロンを種々の時間に処理したところ,PMS注射後45~54時間の0.2mg, 2mg処理で高い排卵率を示した。<BR>4) PMSおよびクロミヘン処理でPMS注射後51時間にコルチゾン•アセテート,コルチコステロン,17α-ハイドロキシ•プロゲステロンを0.2mgおにび2mg処理したが,排卵は全くみられなかった。

著者

ISOLA Michela COSSU Margherita DE LISA Antonello ISOLA Raffaella MASSA Denise CASTI Alberto SOLINAS Paola LANTINI M. Serenella

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.56, no.1, pp.94-97, 2010

被引用文献数

3

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Oxytocin is a cyclic nonapeptide whose best known effects are stimulation of uterine smooth muscle cells during labor and of milk ejection during lactation. Circulating oxytocin originates from the hypothalamus, but its production has also been documented in peripheral tissues. Furthermore, seminal plasma also contains oxytocin, but its functional role is still unknown, although its secretion is generally ascribed to the prostate. In this study, we investigated the possibility that seminal oxytocin is also secreted by other exocrine glands of the human male genital tract. Intramural (Littrè's) glands isolated from bioptic specimens of normal urethrae were processed for immunogold localization of oxytocin. Immunostaining was detected in principal cells, with gold particles specifically found on secretory granules. Basal and endocrine cells were unstained. The present findings suggest that urethral glands not only produce the mucinous layer that protects and lubricates the urethral wall, but also are potential sources of other seminal components, such as oxytocin, which probably play still unclear roles in reproductive physiology.<br>

著者

Jinsha LIU Keiji MOCHIDA Ayumi HASEGAWA Kimiko INOUE Atsuo OGURA

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.2017-148, (Released:2017-12-21)

被引用文献数

5

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Although it is known that the susceptibility of mouse spermatozoa to freezing–thawing varies greatly with genetic background, the underlying mechanisms remain to be elucidated. In this study, to map genetic regions responsible for the susceptibility of spermatozoa to freezing–thawing, we performed in vitro fertilization using spermatozoa from recombinant inbred mice derived from the C57BL/6J and DBA/2J strains, whose spermatozoa showed distinct fertilization abilities after freezing. Genome-wide interval mapping identified two suggestive quantitative trait loci (QTL) associated with fertilization on chromosomes 1 and 11. The strongest QTL on chromosome 11 included 70 genes at 59.237260–61.324742 Mb and another QTL on chromosome 1 included 43 genes at 153.969506–158.217850 Mb. These regions included at least 15 genes involved with testicular expression and possibly with capacitation or sperm motility. Specifically, the Abl2 gene on chromosome 1, which may affect subcellular actin distribution, had polymorphisms between C57BL/6J and DBA/2J that caused at least three amino acid substitutions. A correlation analysis using recombinant inbred strains revealed that the fertilization rate was strongly correlated with the capacitation rate of frozen-thawed spermatozoa after preincubation. This result is consistent with the fact that C57BL/6J frozen–thawed spermatozoa recover their fertilization capacity following treatment with methyl-β-cyclodextrin to enhance sperm capacitation. Thus, our data provide important clues to the molecular mechanisms underlying cryodamage to mouse spermatozoa.

著者

清家 昇

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.38, no.6, pp.j121-j127, 1992

被引用文献数

1 1

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牛胚移植技術の普及を目指し,多くの酪農家と共に種々のETに取り組んだ.非過排卵処置牛からの採卵は供卵牛にホルモン剤を投与しない為、比較的容易に酪農家に受け入れられ,ET技術を理解して頂く上で有効であった.また,高泌乳牛群を用いたETは農家のET技術への偏見を払拭し,ET技術の素晴らしさを再認識して頂く上で効果的であった.特に,2頭のET牛は乳量日本記録を更新し,多くの酪農家の注目を浴びると共に,優良牛を用いてのET普及に大きく拍車をかけた.更に,優良未経産牛群を用いたETは,供卵牛を所有しない一般酪農家のET取り組み方法として有効であると共に,改良の世代間隔を短縮出来る効果的な手段であった.胚の切断2分離技術は,人為的な1卵性双子の作出により,高価な胚を効率的に移植し,回収胚からより多くの産子を獲得できた.またETS子牛の正常性も明らかとなった.

著者

Norikazu MIYASHITA Yasuaki KUBO Miharu YONAI Kanako KANEYAMA Norio SAITO Ken SAWAI Akira MINAMIHASHI Toshiyuki SUZUKI Toshiyuki KOJIMA Takashi NAGAI

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.1108020393-1108020393, (Released:2011-08-05)

被引用文献数

6 13

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Dolly, the first mammal cloned from a somatic cell, had shorter telomeres than age-matched controls and died at an early age because of disease. To investigate longevity and lifetime performance in cloned animals, we produced cloned cows with short telomeres using oviductal epithelial cells as donor cells. At 5 years of age, despite the presence of short telomeres, all cloned cows delivered multiple healthy offspring following artificial insemination with conventionally processed spermatozoa from noncloned bulls, and their milk production was comparable to that of donor cows. Moreover, this study revealed that the offspring had normal-length telomeres in their leukocytes and major organs. Thus, cloned animals have normal functional germ lines, and therefore germ line function can completely restore telomere lengths in clone gametes by telomerase activity, resulting in healthy offspring with normal-length telomeres.

著者

三宅 陽一

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.j111-j118, 1996 (Released:2010-10-20)

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これまでに家畜には多くの染色体異常例があり,それが繁殖障害と密接に関連していることを報告した.今回,それらの結果をもとに,染色体異常例は以下の4つの型に区分されるものと考えた.すなわち,A群は染色体の形態に異常があり相対的不妊症を示すもの(転座,逆位);B群は染色体の形態は正常であるが絶対的不妊症を示すもの(XY female,間性);C群は染色体数の増減があり絶対的不妊症を示すもの(トリソミー,モノソミー);D群は性染色体のキメラが認められ絶対的な不妊症を示すもの(フリーマーチン)である.今後,このような染色体異常例は家畜の遺伝子(群)の機能を解析するうえで,格好のモデルになるものと思われる.

著者

Kudo Toshiyuki Yamamoto Hiroaki Sato Seiji SUTOU Shizuyo

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.42, no.2, pp.101-107, 1996-05-01

参考文献数

28

被引用文献数

1 10

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Aromatase (EC 1.14.14.1.) is a key enzyme of feminizing hormone biosynthesis and catalyzes the conversion of androgens to estrogens. In birds, sex-steroid hormones, especially estrogens, play a critical role in the development of the gonadal glands and the aromatase is one of the most important factors in sex determination. Herein, we cloned the 5' upstream regions of chicken and quail aromatase genes and determined these sequences, which showed high homology between chicken and quail. Moreover, the transcription initiation site of the chicken aromatase gene in early development was determined by the 5'-RACE method. The findings showed that the transcription of the chicken aromatase gene starts from a more upstream site than previously reported.

著者

表記なし

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.52, no.1, pp.143-144, 2006-02-01

著者

橋爪 力 高橋 優子 沼田 恵 SASAKI Koichi UENO Kimiko OHTSUKI Kenji KAWAI Mami ISHII Aya

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.45, no.4, pp.273-281, 1999-08-01

被引用文献数

5 29

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The objectives of this research were to characterize plasma profiles of growth hormone (GH), prolactin (PRL) and insulin-like growth factor-I (IGF-I) during gestation and the lactation period in goats, as well as in their suckling growing kids. There were no obvious changes in the GH or PRL profiles during gestation, but secretion of both GH and PRL increased acutely after parturition. The elevated GH concentrations were maintained until 3 months after parturition, whereas the PRL concentrations decreased gradually after parturition. The elevated hormone concentrations are thought to be caused mainly by an increase in the secretory pulse amplitudes. In contrast to the GH and PRL profiles, the plasma IGF-I concentrations during late gestation were significantly higher than the post-parturition concentrations, and the IGF-I concentrations in late gestation decreased gradually until the day of parturition. The plasma GH, PRL and IGF-I concentrations in kids 1 week after birth were high, and concentrations of each hormone decreased thereafter. These results show that the increased secretory potencies of pituitary GH and PRL after parturition are caused mainly by an increase in the secretory pulse amplitudes and that plasma GH is inversely related to plasma IGF-I compared over gestation and the lactation period in goats.

著者

Korzekwa Anna J. 奥田 潔 Woclawek-Potocka Izabela 村上 周子 SKARZYNSKI Dariusz J

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.52, no.3, pp.353-361, 2006-06-01

被引用文献数

9 54

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We previously showed in <i>in vivo</i> and <i>in vitro</i> studies that nitric oxide (NO) is engaged in luteolysis in cattle. Nitric oxide produced locally in the bovine corpus luteum (CL) inhibits progesterone (P4) synthesis and is suggested to be a component of the luteolytic cascade induced by uterine prostaglandin (PG) F₂_α. In the present study, the molecular mechanisms of NO action during structural luteolysis were studied in cultured bovine luteal cells (Days 15-17 of the estrous cycle). The effects of the NO donor (NONOate; 10^-4M) on DNA fragmentation, cell viability, P4 production and caspase-3 activity were compared with those of PGF₂_α (10^-6M). Moreover, mobilization of intracellular calcium [Ca²⁺]_i and gene expressions of Fas-L, Fas, bcl-2, bax, and caspase-3 in the cells were determined by semi-quantitative RT-PCR after NONOate treatment. Caspase-3 activity was examined calorimetrically. Contrary to PGF₂_α NONOate decreased cell viability. DNA fragmentation after NONOate treatment increased by more than with PGF2₂_α. NONOate increased mobilization of [Ca²⁺]_i in the cells. Although the NO donor did not affect Fas-L and bcl-2 gene expression, it stimulated Fas and bax mRNA and caspase-3 expression. The ratio of bcl-2 to bax mRNA level decreased in the cells treated with NONOate. Moreover, NONOate stimulated caspase-3 activity more effectively than PGF₂_α. The overall results suggest that NO is a luteolytic factor that plays a crucial role in regulation of the estrous cycle in structural luteolysis by inducing apoptosis of luteal cells in cattle.<br>

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

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.