Journal of Reproduction and Development
Online ISSN : 1348-4400
Print ISSN : 0916-8818
ISSN-L : 0916-8818
Advance online publication
Showing 1-28 articles out of 28 articles from Advance online publication
  • Manami SENOO, Hiroshi HOZOJI, Yu ISHIKAWA-YAMAUCHI, Takashi TAKIJIRI, ...
    Article ID: 2020-060
    Published: 2020
    [Advance publication] Released: July 06, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    PTBP1, a well-conserved RNA-binding protein, regulates cellular development by tuning posttranscriptional mRNA modification such as alternative splicing (AS) or mRNA stabilization. We previously revealed that the loss of Ptbp1 in spermatogonia causes the dysregulation of spermatogenesis, but the molecular mechanisms by which PTBP1 regulates spermatogonium homeostasis are unclear. In this study, changes of AS or transcriptome in Ptbp1-knockout (KO) germline stem cells (GSC), an in vitro model of proliferating spermatogonia, was determined by next generation sequencing. We identified more than 200 differentially expressed genes, as well as 85 genes with altered AS due to the loss of PTBP1. Surprisingly, no differentially expressed genes overlapped with different AS genes in Ptbp1-KO GSC. In addition, we observed that the mRNA expression of Nanos3, an essential gene for normal spermatogenesis, was significantly decreased in Ptbp1-KO spermatogonia. We also revealed that PTBP1 protein binds to Nanos3 mRNA in spermatogonia. Furthermore, Nanos3+/−;Ptbp1+/− mice exhibited abnormal spermatogenesis, which resembled the effects of germ cell-specific Ptbp1 KO, whereas no significant abnormality was observed in mice heterozygous for either gene alone. These data implied that PTBP1 regulates alternative splicing and transcriptome in spermatogonia under different molecular pathways, and contributes spermatogenesis, at least in part, in concert with NANOS3.

    Graphical Abstract Fullsize Image
    Download PDF (1753K)
  • Michiko NOGUCHI, Takehito SUZUKI, Reiichiro SATO, Yosuke SASAKI, Kazuy ...
    Article ID: 2020-034
    Published: 2020
    [Advance publication] Released: June 28, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    This study aimed to determine if lactation can be induced by exogenous hormonal treatment in non-pregnant sows. In experiment 1, pseudopregnant animals were divided into four groups and given: 1) 5 mg of estradiol dipropionate (EDP) 5 days before (n = 4), 2) 5 mg of EDP 10 days before (n = 3), 3) 10 mg of EDP 5 days before (n = 3) or 4) 10 mg of EDP 10 days (n = 3) before PGF treatment. Artificial lactation was induced in seven pseudopregnant sows (53.8%) by exogenous hormonal treatment. There was no significant effect of either an increased EDP dosage or interval from the EDP treatment to PGF treatment on the induction rate of artificial lactation. In experiment 2, milk samples were collected from artificial lactating and natural lactating sows (n = 6). IgG and IgA levels in the milk collected from both groups were significantly associated with time during the experimental period. Milk IgG levels 24 h after PGF treatment in artificial lactating sows were higher than those in the colostrum of lactating sows. In experiment 3, hormonal profiles in pseudopregnant sows with (n = 3) or without (n = 3) EDP treatment were determined. There was a significant difference in estradiol-17β levels on days 8, 7 and 5 before PGF treatment between groups. Progesterone and prolactin concentrations did not differ between groups. The present study revealed for the first time that lactation could be induced by exogenous hormonal treatment in non-pregnant sows and that the milk collected from these sows contained high immunoglobulin levels.

    Graphical Abstract Fullsize Image
    Download PDF (1106K)
  • Keisuke KOYAMA, Takayoshi TAKAHASHI
    Article ID: 2020-048
    Published: 2020
    [Advance publication] Released: June 28, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Modern genetic improvement in dairy cattle is directed towards improvement of fertility; however, reproduction traits generally exhibit a genetic antagonism with milk yield. Herein, we aimed to clarify the effects of sire predicted transmitting ability (PTA) for daughter pregnancy rate (DPR) on the reproductive performance and milk yield of daughters in Japanese dairy herds. We conducted a retrospective cohort study on four dairy herds in eastern Hokkaido, Japan, using 1,612 records from 1,018 cows with first, second, or third calvings between March 2015 and September 2018. First, we classified sires into three groups based on the tertile value of their DPR estimate: ≤ −2.2 (low), −2.1 to −0.4 (intermediate), and ≥ −0.3 (high). Subsequently, we compared the sire PTA estimates, reproductive performance, and milk production among DPR groups for each parity of the daughters. In the first and second parity, the hazard of pregnancy by 200 days postpartum was highest in cows from the high-DPR group (P < 0.05); in the third parity, it was unaffected by DPR group. Although sire PTA for milk production in cows from the low-DPR group was highest, actual milk production was unaffected by DPR group regardless of parity. Our findings demonstrate that using sires with PTA for high fertility can enable farmers to improve reproductive performance without decreasing milk yield in Japanese dairy herds. However, it should be noted that sires with PTA for high fertility are at risk for reducing the genetic merit for milk production.

    Graphical Abstract Fullsize Image
    Download PDF (799K)
  • Natsumi ENDO, Larasati Puji RAHAYU, Takashi YAMAMURA, Hitoshi TANAKA, ...
    Article ID: 2019-151
    Published: 2020
    [Advance publication] Released: June 09, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    The objective of the present study was to develop and evaluate a sustained release vaginal progesterone (P4) capsule containing a mixture of mucoadhesive polymer and silicone fluid. Goats were administered a gelatin capsule containing 0.4 g of P4 mixed in silicone fluid and either a hydroxypropylmethylcellulose (HM) or polyaclil starch (PA) base. The mean plasma P4 concentrations at 2 and 12 h after administration were significantly higher in goats treated with PA capsules than in those with HM capsules. The plasma P4 concentrations in goats treated with HM capsules increased and remained above 1.0 ng/ml for 96 h after administration, whereas the plasma P4 concentrations in goats treated with PA capsules remained above 1.0 ng/ml for only 24 h after administration. In the next experiment, an HM capsule was attached to a silicone device and inserted in the vagina for 10 days. The plasma P4 concentration remained similar to that of the natural luteal phase for 9 days. These results suggest that a mixture of mucoadhesive polymer and silicone fluid has the potential to be applied clinically as a sustained release base for estrus synchronization or hormonal therapy.

    Graphical Abstract Fullsize Image
    Download PDF (725K)
  • Rei INOUE, Kana HARADA, Sayaka WAKAYAMA, Masatoshi OOGA, Teruhiko WAKA ...
    Article ID: 2020-059
    Published: 2020
    [Advance publication] Released: June 06, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Mouse oocytes are generally collected after euthanasia. However, if oocytes were collected without euthanasia, then mice could be used to collect oocytes again after recovery. This condition is especially useful for mice that are genotypically rare. In this study, we examined the reusability of mice after collecting oocytes via a surgical operation. When oocytes were collected using medetomidine/midazolam/butorphanol combination anesthesia and examined for the quality of oocytes after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), they could develop to full term at the same rate as controls. When oocytes were collected from those mice a second time, the average number of oocytes was reduced by nearly 1/3. However, the blastocyst and offspring rates of those oocytes after IVF or ICSI were the same as those of the control regardless of the recovery day period. Although GV oocytes can be collected from all reused mice, the final number of offspring did not increase. Interestingly, when oocytes were collected from the front position of the ampulla, 76% of the oviducts possessed oocytes after reuse, and the average number of oocytes significantly increased to a level comparable to that of the control. Finally, we examined whether reused mice can be used as recipient females, and then healthy offspring were obtained similarly as the control recipients. In conclusion, we provide a new method to collect a sufficient number of oocytes from reused mice without concern.

    Graphical Abstract Fullsize Image
    Download PDF (917K)
  • Shota MAYAMA, Nobuhiko HAMAZAKI, Yuki MARUYAMA, Shin MATSUBARA, Atsush ...
    Article ID: 2019-161
    Published: 2020
    [Advance publication] Released: June 05, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Specific gene expression in granulosa cells is key for the function of ovary, but the molecular mechanism of transcriptional activation is not well studied. Here we investigated the regulatory mechanism of the mouse Scd2 gene encoding an enzyme for lipid metabolism. Northern blot and in situ hybridization indicated that the mouse Scd2 mRNA was highly expressed in ovarian granulosa cells. We found four conserved noncoding sequences (CNSs) and two long noncoding RNAs (lncRNAs) transcribed from regions upstream of the Scd2 gene as candidates of regulatory elements/factors. These lncRNAs were predominantly transcribed in the opposite direction to Scd2 and localized in nuclei and showed the correlation with Scd2 expression, raising the possibility of their transcriptional regulatory roles. Indeed, knockdown of both lncRNAs, lncRNA-sc1 and lncRNA-sc2, significantly decreased the Scd2 mRNA level in primary granulosa cells. Then, we investigated the histone modification pattern at this locus by a chromatin immunoprecipitation assay, and two CNSs, CNS1 and CNS2, were found to be marked with high levels of histone H3K9/K27 acetylation in primary granulosa cells. By a reporter gene assay, both CNS1 and CNS2 interdependently exhibited enhancer activity for the Scd2 promoter in primary granulosa cells. These data suggest that the mouse Scd2 gene is activated by two lncRNAs and interdependent enhancers in ovarian granulosa cells, which provides a new insight into transcriptional activation in granulosa cells.

    Graphical Abstract Fullsize Image
    Download PDF (1659K)
  • Yuki HIRADATE, Kenshiro HARA, Kentaro TANEMURA
    Article ID: 2020-002
    Published: 2020
    [Advance publication] Released: June 02, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Previously, we revealed that neurotensin (NTS) derived from the oviduct and uterus can function during fertilization. However, little is known about NTS actions on the pre-implantation embryo after fertilization. Here, we found that pro-Nts mRNA is expressed in the oviduct and uterus during when preimplantation embryos develop and an increase in mRNA level in the uterus is induced by human chorionic gonadotropin (hCG) treatment. Expression of mRNA for two NTS receptors, Ntr1 and Ntr3, was found throughout these stages, whereas Ntr2 mRNA was not detected, suggesting that NTS signaling occurred through NTR1 and NTR3. Supplementation of 1, 10, 100 or 1000 nM NTS to embryo culture medium after fertilization showed that 100 nM NTS significantly improved the blastocyst formation. In comparison, the total number of cells and inner cell mass ratio of blastocysts was not significant different between the 0 nM and 100 nM NTS treatment groups. These results indicate that NTS has a positive effect upon preimplantation embryo development in vitro.

    Download PDF (481K)
  • Irina GARCIA-ISPIERTO, Fernando LÓPEZ-GATIUS
    Article ID: 2020-027
    Published: 2020
    [Advance publication] Released: May 30, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    We examined the effects of a single or 2.5-fold dose of dephereline [a gonadotropin-releasing hormone (GnRH) analogue] as well as the drainage of the smaller follicle at the time of insemination on ovulation in dairy cows with two follicles of pre-ovulatory size in the same ovary. The three study groups included 220 monovular cows (control), 110 non-drained cows with two follicles, and 110 cows with two follicles, of which one was drained. In each group, cows treated with a single dose or 2.5-fold dose of dephereline showed similar results following treatment. Ovulation failure of the non-drained follicle occurred in 29.1% of the drained cows, whereas ovulation occurred in 96% of the non-drained and control cows. Twin pregnancy was recorded in 19.4% of the pregnant non-drained cows with two follicles. In conclusion, the increased dephereline dose did not improve the ovulation rate in any group. Follicular drainage, however, prevented twin pregnancy in cows with two follicles, but also resulted in an increase in the non-drained follicle’s rate of ovulation failure.

    Graphical Abstract Fullsize Image
    Download PDF (982K)
  • Maria SORRENTI, Francesca Gioia KLINGER, Sara IONA, Valerio ROSSI, Ser ...
    Article ID: 2019-141
    Published: 2020
    [Advance publication] Released: May 16, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    In the present work, we described the expression and activity of extracellular signal-related kinases 1-2 (ERK1-2) in mouse primordial germ cells (PGCs) from 8.5–14.5 days post coitum (dpc) and investigated whether these kinases play a role in regulating the various processes of PGC development. Using immunofluorescence and immunoblotting to detect the active phosphorylated form of ERK1-2 (p-ERK1-2), we found that the kinases were present in most proliferating 8.5–10.5 dpc PGCs, low in 11.5 dpc PGCs, and progressively increasing between 12.5–14.5 dpc both in female and male PGCs. In vitro culture experiments showed that inhibiting activation of ERK1-2 with the MEK-specific inhibitor U0126 significantly reduced the growth of 8.5 dpc PGCs in culture but had little effect on 11.5–12.5 dpc PGCs. Moreover, we found that the inhibitor did not affect the adhesion of 11.5 dpc PGCs, but it significantly reduced their motility features onto a cell monolayer. Further, while the ability of female PGCs to begin meiosis was not significantly affected by U0126, their progression through meiotic prophase I was slowed down. Notably, the activity of ERK1-2 was necessary for maintaining the correct expression of oocyte-specific genes crucial for germ cells survival and the formation of primordial follicles.

    Graphical Abstract Fullsize Image
    Download PDF (1797K)
  • Daiki SHIKATA, Takuto YAMAMOTO, Shinnosuke HONDA, Shuntaro IKEDA, Naoj ...
    Article ID: 2020-036
    Published: 2020
    [Advance publication] Released: May 06, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Maintaining genomic integrity in mammalian early embryos, which are deficient in DNA damage repair, is critical for normal preimplantation and subsequent development. Abnormalities in DNA damage repair in preimplantation embryos can cause not only developmental arrest, but also diseases such as congenital disorders and cancers. Histone H4 lysine 20 monomethylation (H4K20me1) is involved in DNA damage repair and regulation of gene expression. However, little is known about the role of H4K20me1 during mouse preimplantation development. In this study, we revealed that H4K20me1 mediated by SETD8 is involved in maintaining genomic integrity. H4K20me1 was present throughout preimplantation development. In addition, reduction in the level of H4K20me1 by inhibition of SETD8 activity or a dominant-negative mutant of histone H4 resulted in developmental arrest at the S/G2 phase and excessive accumulation of DNA double-strand breaks. Together, our results suggest that H4K20me1, a type of epigenetic modification, is associated with the maintenance of genomic integrity and is essential for preimplantation development. A better understanding of the mechanisms involved in maintaining genome integrity during preimplantation development could contribute to advances in reproductive medicine and technology.

    Graphical Abstract Fullsize Image
    Download PDF (1086K)
  • Sevastiani ANTONOULI, Maria Grazia PALMERINI, Serena BIANCHI, Gianna R ...
    Article ID: 2019-147
    Published: 2020
    [Advance publication] Released: April 28, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Controlled ovarian hyperstimulation (COH) is routinary used in assisted reproductive technologies (ARTs) to increase the yields of mature oocytes. The possibility that patients with a history of failures or poor-responders may develop side-effects following these treatments is still debated. Epidemiological studies reported controversial results about pregnancy outcome and the risk of developing gynecological cancers. By using a mouse model, here we compared the ultrastructural features of Fallopian tubes (FTs) obtained from mice undergoing or not (control, CTR) four (4R) and eight (8R) rounds of gonadotropin stimulation. Although the morphological characteristics of oviductal layers seemed unaffected by repeated treatments, dose-response ultrastructural alterations in the ampulla appeared in the 4R group and even more in the 8R group. The targets were oviductal ciliated (CCs) and non-ciliated (NCCs) cells, which showed damaged mitochondria and glycogen accumulations in the cytoplasm. The drastic reduction of CCs, evident after 4R, was supported by the absence of cilia. After 8R, glycogen granules were significantly reduced and massive degeneration of mitochondria, which appeared swollen and/or vacuolated, occurred in NCCs. Moreover, disintegrated mitochondria were found at the periphery of mitophagic vacuoles with evident signs of cristolysis. The morphometric analysis evidenced a significant increase in the density and frequency of damaged mitochondria after 4R and 8R. The absence of cilia, necessary to sustain oviductal transport of oocytes, spermatozoa and embryos, may originate from either mitochondrial dysfunction or glycogen consumption. These results suggest that repeated COH treatments could induce alterations impairing fertilization and embryo transport toward the uterus.

    Graphical Abstract Fullsize Image
    Download PDF (1992K)
  • Shiori MINABE, Sho NAKAMURA, Eri FUKUSHIMA, Marimo SATO, Kana IKEGAMI, ...
    Article ID: 2019-164
    Published: 2020
    [Advance publication] Released: April 26, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Accumulating evidence suggests that kisspeptin-GPR54 signaling is indispensable for gonadotropin-releasing hormone (GnRH)/gonadotropin secretion and consequent reproductive functions in mammals. Conventional Kiss1 knockout (KO) mice and rats are reported to be infertile. To date, however, no study has investigated the effect of inducible central Kiss1 KO/knockdown on pulsatile gonadotropin release in male mammals. Here we report an in vivo analysis of inducible conditional Kiss1 knockdown male mice. The mice were generated by a bilateral injections of either adeno-associated virus (AAV) vectors driving Cre recombinase (AAV-Cre) or AAV vectors driving GFP (AAV-GFP, control) into the hypothalamic arcuate nucleus (ARC) of Kiss1-floxed male mice, in which exon 3 of the Kiss1 gene were floxed with loxP sites. Four weeks after the AAV-Cre injection, the mice showed a profound decrease in the both number of ARC Kiss1-expressing cells and the luteinizing hormone (LH) pulse frequency. Interestingly, pulsatile LH secretion was apparent 8 weeks after the AAV-Cre injection despite the suppression of ARC Kiss1 expression. The control Kiss1-floxed mice infected with AAV-GFP showed apparent LH pulses and Kiss1 expression in the ARC at both 4 and 8 weeks after the AAV-GFP injection. These results with an inducible conditional Kiss1 knockdown in the ARC of male mice suggest that ARC kisspeptin neurons are responsible for pulsatile LH secretion in male mice, and indicate the possibility of a compensatory mechanism that restores GnRH/LH pulse generation.

    Graphical Abstract Fullsize Image
    Download PDF (4625K)
  • Ho-Geun JEGAL, Hyo-Jin PARK, Jin-Woo KIM, Seul-Gi YANG, Min-Ji KIM, De ...
    Article ID: 2020-013
    Published: 2020
    [Advance publication] Released: April 22, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Ruthenium red (RR) inhibits calcium (Ca2+) entry from the cytoplasm to the mitochondria, and is involved in maintenance of Ca2+ homeostasis in mammalian cells. Ca2+ homeostasis is very important for further embryonic development of fertilized oocytes. However, the effect of RR on mitochondria-Ca2+ (mito-Ca2+) levels during in vitro fertilization (IVF) on subsequent blastocyst developmental capacity in porcine is unclear. The present study explored the regulation of mito-Ca2+ levels using RR and/or histamine in fertilized oocytes and their influence on blastocyst developmental capacity in pigs. Red fluorescence intensity by the mito-Ca2+ detection dye Rhod-2 was significantly increased (P < 0.05) in zygotes 6 h after IVF compared to mature oocytes. Based on these results, we investigated the changes in mito-Ca2+ by RR (10 and 20 μM) in presumptive zygotes using Rhod-2 staining and mito-Ca2+ uptake 1 (MICU1) protein levels as an indicator of mito-Ca2+ uptake using western blot analysis. As expected, RR-treated zygotes displayed decreased protein levels of MICU1 and Rhod-2 red fluorescence intensity compared to non-treated zygotes 6 h after IVF. Blastocyst development rate of 20 μM RR-treated zygotes was significantly increased 6 h after IVF (P < 0.05) due to improved mitochondrial functions. Conversely, the blastocyst development rate was significantly decreased in histamine (mito-Ca2+ inhibitor, 100 nM) treated zygotes (P < 0.05). The collective results demonstrate that RR improves blastocyst development in porcine embryos by regulating mito-Ca2+ and MICU1 expression following IVF.

    Graphical Abstract Fullsize Image
    Download PDF (2232K)
  • Kana IKEGAMI, Teppei GOTO, Sho NAKAMURA, Youki WATANABE, Arisa SUGIMOT ...
    Article ID: 2020-026
    Published: 2020
    [Advance publication] Released: April 20, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    The present study aimed to evaluate whether novel conditional kisspeptin neuron-specific Kiss1 knockout (KO) mice utilizing the Cre-loxP system could recapitulate the infertility of global Kiss1 KO models, thereby providing further evidence for the fundamental role of hypothalamic kisspeptin neurons in regulating mammalian reproduction. We generated Kiss1-floxed mice and hypothalamic kisspeptin neuron-specific Cre-expressing transgenic mice and then crossed these two lines. The conditional Kiss1 KO mice showed pubertal failure along with a suppression of gonadotropin secretion and ovarian atrophy. These results indicate that newly-created hypothalamic Kiss1 KO mice obtained by the Cre-loxP system recapitulated the infertility of global Kiss1 KO models, suggesting that hypothalamic kisspeptin, but not peripheral kisspeptin, is critical for reproduction. Importantly, these Kiss1-floxed mice are now available and will be a valuable tool for detailed analyses of roles of each population of kisspeptin neurons in the brain and peripheral kisspeptin-producing cells by the spatiotemporal-specific manipulation of Cre expression.

    Graphical Abstract Fullsize Image
    Graphical Abstract
    Download PDF (3640K)
  • Keiji MOCHIDA
    Article ID: 2020-033
    Published: 2020
    [Advance publication] Released: April 19, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Assisted reproductive technologies (ARTs) are widely used in the animal industry, human clinics, and for basic research. In small laboratory animal species such as mice, ARTs are essential for the production of animals for experiments, the preservation of genetic resources, and for the generation of new strains of genetically modified animals. The RIKEN BioResource Research Center (BRC) is one of the largest repositories of such animal bioresources, and maintains approximately 9,500 strains of mice with a variety of genetic backgrounds. We have sought to devise ARTs specific to the reproductive and physiological characteristics of each strain. Such ARTs include superovulation, in vitro fertilization (IVF), the cryopreservation of embryos and spermatozoa, transportation of cryopreserved materials and embryo transfer (ET). Of these, superovulation likely has the most influence on animal production because it determines the quantity of starting material for other ARTs. Superovulation using anti-inhibin serum combined with estrous synchronization has resulted in approximately a three-fold increase in production efficiency with IVF–ET in the C57BL/6J strain. Wild-derived strains are important as genetically diverse resources for murine rodents (Genus Mus), and many are unique to the BRC. We have also successfully developed ARTs for more than 50 wild-derived strains, which have been cryopreserved for future use. Our work to improve and develop ARTs for mice and other small laboratory species will contribute to the cost-effectiveness of routine operations at repository centers, and to the provision of high quality animals for research use.

    Graphical Abstract Fullsize Image
    Download PDF (673K)
  • Takuya SASAKI, Tomoya SONODA, Ryoki TATEBAYASHI, Yuri KITAGAWA, Shinya ...
    Article ID: 2019-145
    Published: 2020
    [Advance publication] Released: April 12, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Accumulating evidence suggests that KNDy neurons located in the hypothalamic arcuate nucleus (ARC), which are reported to express kisspeptin, neurokinin B, and dynorphin A, are indispensable for the gonadotropin-releasing hormone (GnRH) pulse generation that results in rhythmic GnRH secretion. The aims of the present study were to investigate the effects of peripheral administration of the neurokinin 3 receptor (NK3R/TACR3, a receptor for neurokinin B) antagonist, SB223412, on GnRH pulse-generating activity and pulsatile luteinizing hormone (LH) secretion in ovariectomized Shiba goats treated with luteal phase levels of estrogen. The NK3R antagonist was infused intravenously for 4 h {0.16 or 1.6 mg/(kg body weight [BW]·4 h)} during which multiple unit activity (MUA) in the ARC was recorded, an electrophysiological technique commonly employed to monitor GnRH pulse generator activity. In a separate experiment, the NK3R antagonist (40 or 200 mg/[kg BW·day]) was administered orally for 7 days to determine whether the NK3R antagonist could modulate pulsatile LH secretion when administered via the oral route. Intravenous infusion of the NK3R antagonist significantly increased the interval of episodic bursts of MUA compared with that of the controls. Oral administration of the antagonist for 7 days also significantly prolonged the interpulse interval of LH pulses. The results of this study demonstrate that peripheral administration of an NK3R antagonist suppresses pulsatile LH secretion by acting on the GnRH pulse generator, suggesting that NK3R antagonist administration could be used to modulate reproductive functions in ruminants.

    Graphical Abstract Fullsize Image
    Download PDF (1545K)
  • Mingming LEI, Rong CHEN, Qingming QING, Huanxi ZHU, Zhendan SHI
    Article ID: 2019-110
    Published: 2020
    [Advance publication] Released: April 11, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Magang geese exhibit a unique characteristic of follicular development, with eight largest orderly arranged pre-ovulatory follicles in the abdominal cavity. However, little is known about the mechanisms underlying these this follicular development. This study aimed to compare gene expression profiles of granulosa cells (GCs) at different stages of follicular development and provide comprehensive insights into follicle selection and the mechanisms underlying the well-defined follicle hierarchy in Magang geese. GCs of large white follicles (LWFs), small yellow follicles (SYFs), F8, F4, and F1 were used for RNA-seq analysis; 374, 1117, 791, and 593 genes were differentially expressed in stages LWFs to SYFs, SYFs to F8, F8 to F4, and F4 to F1, respectively, suggesting that these genes contribute to follicle selection and development. Reliability of sequencing data was verified through qPCR analysis of 24 genes. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways revealed a complex mechanism that remodels the extracellular matrix and turnover of extracellular matrix components in follicular development and ovulation and involves multiple pathway, such as focal adhesion, adherens junction, and extracellular matrix–receptor interaction. Some unique characteristics were observed during the different follicular development stages. For instance, some differentially expressed genes were enriched in progesterone-mediated oocyte maturation and steroid biosynthesis from stage SYFs to F8, whereas others were enriched in actin cytoskeleton regulation and vascular smooth muscle contraction from stage F4 to F1. These findings enhance our current understanding of GC function and ovarian follicles during the key stages of follicular development.

    Graphical Abstract Fullsize Image
    Download PDF (1023K)
  • Fumie MAGATA
    Article ID: 2020-021
    Published: 2020
    [Advance publication] Released: April 11, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Uterine inflammatory diseases commonly occur in postpartum dairy cows, resulting in reduced reproductive performance due to aberrant uterine and ovarian activity. Infection of the uterus with gram-negative bacteria results in the detection of lipopolysaccharide (LPS) in the plasma and follicular fluid of cows along with uterine inflammation. LPS acts on follicular components such as theca cells, granulosa cells, and follicle-enclosed oocytes, leading to impaired follicular activity. Follicles with a high LPS environment exhibit reduced follicular steroidogenesis due to the inhibition of steroidogenic enzyme transcription. Primary cell cultures of bovine granulosa and theca cells have shown that LPS acts on follicular cells to impair steroid production, which may disturb follicle growth and/or reduce their ability to ovulate. Even if ovulation occurs, cows with uterine inflammation are less likely to conceive because in addition to uterine damage, LPS also impairs the developmental competence of oocytes. LPS perturbs the nuclear and cytoplasmic maturation of bovine oocytes. Moreover, oocytes matured in an LPS treatment are less likely to develop into the blastocyst stage. Such oocytes also have a reduced number of trophoblast cells in blastocysts. Therefore, the detrimental effects of LPS on ovarian activity may be partly responsible for infertility in cows with uterine inflammation. Novel treatment and prevention strategies for uterine inflammatory diseases can be developed by advancing our knowledge of the pathophysiology underlying ovarian dysfunction, and this can only be achieved by further research. The present review outlines the molecular pathogenesis of LPS-induced ovarian dysfunction.

    Graphical Abstract Fullsize Image
    Download PDF (659K)
  • Fernando LÓPEZ-GATIUS, Irina GARCIA-ISPIERTO
    Article ID: 2020-014
    Published: 2020
    [Advance publication] Released: April 07, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    In this study, we present two proposed approaches to prevent twin pregnancies in dairy cattle: 1) single, in vitro-produced embryonic transfer into a recipient cow or 2) subordinate follicle drainage at the time of insemination. Both procedures lead to improved embryonic survival. As the use of sexed semen generates herd replacements and additional heifers, we propose the transfer of a single female cattle embryo into cows that are not suitable for producing replacements, and follicular drainage in lactating cows with genetic merit. This should eliminate economic losses associated with twin pregnancies and increase cattle output of the herd.

    Download PDF (469K)
  • Hiroya KADOKAWA
    Article ID: 2020-012
    Published: 2020
    [Advance publication] Released: April 06, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Previous studies in the 1960s and 1970s have reported that both gonadotropin-releasing hormone (GnRH) and estradiol-activated nuclear estrogen receptors regulate gonadotropin secretion in women. However, I had previously reported that gonadotroph function is regulated by complex crosstalk between several membrane receptors. RNA-seq had previously revealed 259 different receptor genes expressed in the anterior pituitary of heifers. However, the biological roles of most of these receptors remain unknown. I identified four new receptors of interest: G protein-coupled receptor 30 (GPR30), anti-Mullerian hormone (AMH) receptor type 2 (AMHR2), and G protein-coupled receptors 61 and 153 (GPR61 and GPR153). GPR30 rapidly (within a few minutes) mediates picomolar, but not nanomolar, levels of estradiol to suppress GnRH-induced luteinizing hormone (LH) secretion from bovine gonadotrophs, without decreasing mRNA expressions of the LHα, LHβ, or follicle-stimulating hormone (FSH) β subunits. GPR30 is activated by other endogenous estrogens, estrone and estriol. Moreover, GPR30 activation by zearalenone, a nonsteroidal mycoestrogen, suppresses LH secretion. AMHR2, activated by AMH, stimulates LH and FSH secretion, thus regulating gonadotrophs, where other TGF-β family members, including inhibin and activin, potentially affect FSH secretion. I also show that GPR61, activated by its ligand (recently discovered) significantly alters LH and FSH secretion. GPR61, GPR153, and AMHR2 co-localize with the GnRH receptor in unevenly dispersed areas of the bovine gonadotroph cell surface, probably lipid rafts. The findings summarized in this review reveal a new paradigm regarding the mechanisms regulating reproduction via novel receptors expressed on bovine gonadotrophs.

    Graphical Abstract Fullsize Image
    Download PDF (3035K)
  • Ryo NISHIMURA, Kiyoshi OKUDA
    Article ID: 2020-018
    Published: 2020
    [Advance publication] Released: April 04, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    There has been increasing interest in the role of hypoxia in the microenvironment of organs, because of the discovery of hypoxia-inducible factor-1 (HIF1), which acts as a transcription factor for many genes activated specifically under hypoxic conditions. The ovary changes day by day during the estrous cycle as it goes through phases of follicular growth, ovulation, and formation and regression of the corpus luteum (CL). These phenomena are regulated by hypothalamic and pituitary hormones, sex steroids, peptides and cytokines, as well as oxygen conditions. Hypoxia strongly induces angiogenesis via transcription of a potent angiogenic factor, vascular endothelial growth factor (VEGF), that is regulated by HIF1. A CL forms with a rapid increase of angiogenesis that is mainly induced by HIF1-VEGF signaling. Hypoxia also contributes to luteolysis by down-regulating progesterone synthesis and by up-regulating apoptosis of luteal cells. This review focuses on recent studies on the roles of hypoxia- and HIF1-regulated genes in the regulation of bovine CL function.

    Graphical Abstract Fullsize Image
    Download PDF (315K)
  • Mito KANATSU-SHINOHARA, Guiying CHEN, Hiroko MORIMOTO, Takashi SHINOHA ...
    Article ID: 2020-019
    Published: 2020
    [Advance publication] Released: March 26, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    The spermatogonial stem cell (SSC) population in testis is small, and the lack of SSC markers has severely handicapped research on these cells. During our attempt to identify genes involved in SSC aging, we found that CD2 is expressed in cultured SSCs. Flow cytometric analysis and spermatogonial transplantation experiments showed that CD2 is expressed in SSCs from mature adult mouse testes. Cultured SSCs transfected with short hairpin RNAs (shRNAs) against CD2 proliferated poorly and showed an increased frequency of apoptosis. Moreover, functional analysis of transfected cells revealed impairment of SSC activity. Fluorescence activated cell sorting and spermatogonial transplantation experiments showed that CD2 is expressed not only in mouse but also in rat SSCs. The results indicate that CD2 is a novel SSC surface marker conserved between mouse and rat SSCs.

    Graphical Abstract Fullsize Image
    Download PDF (13919K)
  • Felix R. GRAUBNER, Miguel Tavares PEREIRA, Alois BOOS, Mariusz P. KOWA ...
    Article ID: 2019-157
    Published: 2020
    [Advance publication] Released: March 20, 2020
    JOURNALS OPEN ACCESS ADVANCE PUBLICATION

    Recently, we established an in vitro model with immortalized dog uterine stromal (DUS) cells for investigations into canine-specific decidualization. Their capability to decidualize was assessed with cAMP- and PGE2. Here, we show that the effects of PGE2 are mediated through both of the cAMP-mediating PGE2 receptors (PTGER2/4). Their functional inhibition suppressed gene expression of PRLR and PGR in DUS cells. We also assessed the effects of cAMP and PGE2 on selected extracellular matrix components and CX43, and showed that cAMP, but not PGE2, increases COL4, ECM1 and CX43 protein levels during in vitro decidualization, indicating a mesenchymal-epithelial decidual transformation in these cells. Thus, although PGE2 is involved in decidualization, it does not appear to regulate extracellular matrix. Further, the role of progesterone (P4) during in vitro decidualization was addressed. P4 upregulated PRLR and PGR in DUS cells, but these effects were not influenced by PGE2; both P4 and PGE2 hormones appeared to act independently. P4 did not affect IGF1 expression, which was upregulated by PGE2, however, it suppressed expression of IGF2, also in the presence of PGE2. Similarly, P4 did not affect PGE2 synthase (PTGES), but in the presence of PGE2 it increased PTGER2 levels and, regardless of the presence of PGE2, suppressed expression of PTGER4. Our results indicate a reciprocal regulatory loop between PGE2 and P4 during canine in vitro decidualization: whereas P4 may be involved in regulating PGE2-mediated decidualization by regulating the availability of its receptors, PGE2 regulates PGR levels in a manner dependent on PTGER2 and -4.

    Graphical Abstract Fullsize Image
    Graphical Abstract
    Download PDF (1241K)
  • Maajid Hassan BHAT, Syed Hilal YAQOOB, Firdous Ahmad KHAN, Hilal Musad ...
    Article ID: 2018-126e
    Published: 2019
    [Advance publication] Released: May 15, 2019
    JOURNALS FREE ACCESS ADVANCE PUBLICATION

    This article released online on March 5, 2019 as advance publication was withdrawn from consideration for publication in Journal of Reproduction and Development at author’s request.

    Download PDF (78K)
  • Adeleh ZABIHI, Hamed Karami SHABANKAREH, Hadi HAJARIAN, Saheb FOROUTAN ...
    Article ID: 2018-102e
    Published: 2019
    [Advance publication] Released: March 28, 2019
    JOURNALS FREE ACCESS ADVANCE PUBLICATION

    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.

    Download PDF (86K)
  • Maajid Hassan BHAT, Syed Hilal YAQOOB, Firdous Ahmad KHAN, Hilal Musad ...
    Article ID: 2018-126
    Published: 2019
    [Advance publication] Released: March 05, 2019
    JOURNALS FREE ACCESS ADVANCE PUBLICATION
    This article released online on March 5, 2019 as advance publication was withdrawn from consideration for publication in Journal of Reproduction and Development at author’s request.
    Download PDF (356K)
  • Adeleh ZABIHI, Hamed Karami SHABANKAREH, Hadi HAJARIAN, Saheb FOROUTAN ...
    Article ID: 2018-102
    Published: 2019
    [Advance publication] Released: January 18, 2019
    JOURNALS FREE ACCESS ADVANCE PUBLICATION
    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.
    Download PDF (622K)
  • Charlie HUVENEERS, Nicholas M. OTWAY, Megan T. STORRIE, Robert G. HARC ...
    Article ID: 20144
    Published: 2009
    [Advance publication] Released: February 23, 2009
    JOURNALS FREE ACCESS ADVANCE PUBLICATION
    This article was retracted. See the Notification.
    Download PDF (661K)
feedback
Top