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Journal of Reproduction and Development

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https://www.readbyqxmd.com/read/27904015/deletion-of-conserved-sequences-in-ig-dmr-at-dlk1-gtl2-locus-suggests-their-involvement-in-expression-of-paternally-expressed-genes-in-mice
#1
Takeshi Saito, Satoshi Hara, Moe Tamano, Hiroshi Asahara, Shuji Takada
Expression regulation of the Dlk1-Dio3 imprinted domain by the intergenic differentially methylated region (IG-DMR) is essential for normal embryonic development in mammals. In this study, we investigated conserved IG-DMR genomic sequences in eutherians to elucidate their role in genomic imprinting of the Dlk1-Dio3 domain. Using a comparative genomics approach, we identified three highly conserved sequences in IG-DMR. To elucidate the functions of these sequences in vivo, we generated mutant mice lacking each of the identified highly conserved sequences using the CRISPR/Cas9 system...
December 1, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27904014/male-infertility-related-molecules-involved-in-sperm-oocyte-fusion
#2
Lisha Mou, Ni Xie
Male infertility has become a very serious problem in the human reproduction system, but the molecular mechanism of infertility remains largely unknown. Fertilization is the phenomenon in which a sperm and oocyte find each other, interact, and fuse. Sperm-oocyte fusion-related factors on the sperm side play crucial roles in male infertility. For example, IZUMO1 is well-known as a sperm protein essential for fusion of a sperm and oocyte, but its dysfunction or mutation can result in male infertility. Recent studies showed a novel sperm protein named Bactericidal/permeability-increasing protein (BPI), which takes part in the sperm-oocyte fusion process...
December 1, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27890874/development-of-a-lipopolysaccharide-lps-supplemented-adjuvant-and-its-effects-on-cell-mediated-and-humoral-immune-responses-in-male-rats-immunized-against-sperm
#3
Junko Noguchi, Shinya Watanabe, Thanh Q Dang Nguyen, Kazuhiro Kikuchi, Hiroyuki Kaneko
Supplementation with lipopolysaccharide (LPS) from non-pathogenic Escherichia coli was found to enhance the adjuvant effects of a veterinary vaccine adjuvant (ISA 71VG(®)). Sperm immunization using 71VG as an adjuvant in the immature period induced infertility in 25% of male rats, whereas this increased to 62.5% after immunization with 71VG + LPS or Freund's complete adjuvant (FCA). Mean testicular weight of non-sterile males in the 71VG + LPS group was significantly lower than that in the 71VG or FCA group...
November 28, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27890873/overabundance-of-sika-deer-and-immunocontraception
#4
Junko Noguchi
The impact of deer overabundance is a worldwide problem. Along with habitat expansion and population increase, damage by sika deer to the forest ecosystem and agriculture has become a serious issue in Japan. Deer also transmit a number of diseases and parasites to humans and livestock. The overabundance of deer is a result of their strong fecundity, and therefore the present situation should, in theory, be tackled by experts in reproductive biology.
November 28, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27867162/a-unique-mechanism-regulating-gene-expression-in-1-cell-embryos
#5
Ryoma Yamamoto, Fugaku Aoki
After fertilization, the genome of zygotes is transcriptionally silent. The timing of the initiation of transcription is species-specific and occurs at the mid-1-cell stage in mice. Recent analyses using high-throughput sequencing (HTS) have identified thousands of genes transcribed at the 1-cell stage, and the pattern of expression among these genes appears to be unique. In this article, we show the result of an additional analysis using HTS data from a previous study, and present the hypothesis that an extremely loose chromatin structure causes promiscuous gene expression in 1-cell embryos...
November 18, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27867161/departure-from-optimal-o2-level-for-mouse-trophoblast-stem-cell-proliferation-and-potency-leads-to-most-rapid-ampk-activation
#6
Yu Yang, Zhongliang Jiang, Alan Bolnick, Jing Dai, Elizabeth E Puscheck, Daniel A Rappolee
Previous studies showed that cultured mouse trophoblast stem cells (mTSCs) have the most rapid proliferation, normal maintenance of stemness/potency, the least spontaneous differentiation, and the lowest level of stress-activated protein kinase (SAPK) when incubated at 2% O2 rather than at the traditional 20% O2 or hypoxic (0.5% and 0% O2) conditions. Switching from 2% O2 induced fast SAPK responses. Here we tested the dose response of AMP-activated protein kinase (AMPK) in its active form (pAMPK Thr172P) at O2 levels from 20-0%, and also tested whether pAMPK levels show similar rapid changes when mTSC cultures were switched from the optimal 2% O2 to other O2 conditions...
November 18, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27853050/long-term-changes-in-plasma-anti-m%C3%A3-llerian-hormone-concentration-and-the-relationship-with-superovulatory-response-in-japanese-black-cattle
#7
Hiroki Hirayama, Akira Naito, Shigeo Fukuda, Takashi Fujii, Masatsugu Asada, Yasushi Inaba, Toshiro Takedomi, Masakazu Kawamata, Satoru Moriyasu, Soichi Kageyama
The concentration of circulating anti-Müllerian hormone (AMH) in cattle is a useful endocrine marker for ovarian response to superovulation. Although the AMH concentration undergoes little variation throughout the estrous cycle, its long-term changes remain incompletely understood. Here, we investigated the relationship between superovulation response and plasma AMH concentration in Japanese Black cattle and the long-term changes in plasma AMH concentration of embryo donor cows and heifers. The median, 25th percentile, and 75th percentile of AMH concentrations in 222 mature animals were 0...
November 17, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27840375/hypoxia-inducible-factor-1-mediates-hypoxia-enhanced-synthesis-of-progesterone-during-luteinization-of-granulosa-cells
#8
Fadhillah, Shin Yoshioka, Ryo Nishimura, Yuki Yamamoto, Koji Kimura, Kiyoshi Okuda
Hypoxia has been suggested to enhance progesterone (P4) synthesis in luteinizing granulosa cells (GCs), but the mechanism is unclear. The present study was designed to test the hypothesis that the hypoxia-induced increase in P4 synthesis during luteinization in bovine GCs is mediated by hypoxia-inducible factor 1 (HIF-1). GCs obtained from small antral follicles were cultured with 2 µg/ml insulin in combination with 10 µM forskolin for 24 h as a model of luteinizing GCs. To examine the influence of HIF-1 on P4 synthesis, we determined the effect of changes in protein expression of the α-subunit of HIF-1 (HIF1A) on P4 production and on the expression levels of StAR, P450scc, and 3β-HSD...
November 11, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27795465/improvement-of-the-developmental-competence-of-porcine-oocytes-collected-from-early-antral-follicles-by-cytoplast-fusion
#9
Thanh Quang Dang-Nguyen, Ruth Appeltant, Tamas Somfai, Shinya Ishihara, Nguyen Thi Men, Elisa Caroline da Silva Santos, Junko Noguchi, Hiroyuki Kaneko, Kazuhiro Kikuchi
In the present study, we propose an alternative technique called cytoplast fusion to improve the maturation rate and developmental competence of growing oocytes collected from early antral follicles in pigs. We examined whether the fusion of a growing oocyte with the cytoplast from a fully-grown oocyte (CFR group) could better promote maturation and developmental competence of the growing oocyte compared to germinal vesicle (GV) transfer (GVTR group). After 44 h of in vitro maturation (IVM), most growing oocytes (GR group) were still arrested at the GV stage (64...
October 29, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27773888/%C3%AE-aminobutyric-acid-suppresses-enhancement-of-hamster-sperm-hyperactivation-by-5-hydroxytryptamine
#10
Masakatsu Fujinoki, Gen L Takei
Sperm hyperactivation is regulated by hormones present in the oviduct. In hamsters, 5-hydroxytryptamine (5HT) enhances hyperactivation associated with the 5HT2 receptor and 5HT4 receptor, while 17β-estradiol (E2) and γ-aminobutyric acid (GABA) suppress the association of the estrogen receptor and GABAA receptor, respectively. In the present study, we examined the regulatory interactions among 5HT, GABA, and E2 in the regulation of hamster sperm hyperactivation. When sperm were exposed to E2 prior to 5HT exposure, E2 did not affect 5HT-enhanced hyperactivation...
October 24, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27773885/involvement-of-dna-methylation-in-regulating-rat-prop1-gene-expression-during-pituitary-organogenesis
#11
Hiroto Nishihara, Saishu Yoshida, Naoko Kanno, Naoto Nishimura, Hiroki Ueharu, Jun Ohgane, Takako Kato, Yukio Kato
PROP1 is a pituitary specific transcription factor that plays a crucial role in pituitary organogenesis. The Prop1 shows varied expression patterns that promptly emerge and then fade during the early embryonic period. However, the regulatory mechanisms governing Prop1 expression remain unclear. Here, we investigated whether Prop1 was under epigenetic regulation by DNA methylation. Bisulfite sequencing was performed on DNA obtained from the pituitary glands and livers of rats on embryonic days (E) 13.5 and E14...
October 21, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27773884/efficient-mutagenesis-by-crispr-cas-system-during-meiotic-maturation-of-porcine-oocytes
#12
Asuka Onuma, Wataru Fujii, Koji Sugiura, Kunihiko Naito
Genome editing using the CRISPR/Cas system can induce mutations with high efficiency, and allows easier production of genome-modified animals than that offered by the conventional method where embryonic stem cells are used. However, studies using CRISPR/Cas systems have been mostly limited to proliferating somatic cells and pronuclear-stage fertilized eggs. In contrast, the efficiency of a CRISPR/Cas system in immature and maturing oocytes progressing through meiosis has not yet been assessed. In the present study, we evaluated the genome-modification efficiency of the CRISPR/Cas system during meiotic maturation of porcine oocytes...
October 21, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27746409/an-immortalized-steroidogenic-goat-granulosa-cell-line-as-a-model-system-to-study-the-effect-of-the-endoplasmic-reticulum-er-stress-response-on-steroidogenesis
#13
Diqi Yang, Lei Wang, Pengfei Lin, Tingting Jiang, Nan Wang, Fan Zhao, Huatao Chen, Keqiong Tang, Dong Zhou, Aihua Wang, Yaping Jin
With granulosa and theca cells, the ovaries are responsible for producing oocytes and secreting sex steroids such as estrogen and progesterone. Endoplasmic reticulum stress (ERS) plays an important role in follicle atresia and embryo implantation. In this study, goat granulosa cells were isolated from medium-sized (4-6 mm) healthy follicles. Primary granulosa cells were immortalized by transfection with human telomerase reverse transcriptase (hTERT) to establish a goat granulosa cell line (hTERT-GGCs). These hTERT-GGCs expressed hTERT and had relatively long telomeres at passage 50...
October 15, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27725344/production-of-%C3%AE-1-3-galactosyltransferase-and-cytidine-monophosphate-n-acetylneuraminic-acid-hydroxylase-gene-double-deficient-pigs-by-crispr-cas9-and-handmade-cloning
#14
Hanchao Gao, Chengjiang Zhao, Xi Xiang, Yong Li, Yanli Zhao, Zesong Li, Dengke Pan, Yifan Dai, Hidetaka Hara, David K C Cooper, Zhiming Cai, Lisha Mou
Gene-knockout pigs hold great promise as a solution to the shortage of organs from donor animals for xenotransplantation. Several groups have generated gene-knockout pigs via clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) and somatic cell nuclear transfer (SCNT). Herein, we adopted a simple and micromanipulator-free method, handmade cloning (HMC) instead of SCNT, to generate double gene-knockout pigs. First, we applied the CRISPR/Cas9 system to target α1,3-galactosyltransferase (GGTA1) and cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes simultaneously in porcine fetal fibroblast cells (PFFs), which were derived from wild-type Chinese domestic miniature Wuzhishan pigs...
October 8, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27725347/lack-of-calcium-oscillation-causes-failure-of-oocyte-activation-after-intracytoplasmic-sperm-injection-in-pigs
#15
Michiko Nakai, Junya Ito, Shun-Ichi Suzuki, Dai-Ichiro Fuchimoto, Shoichiro Senbon, Misae Suzuki, Junko Noguchi, Hiroyuki Kaneko, Akira Onishi, Naomi Kashiwazaki, Kazuhiro Kikuchi
In pigs, the efficiency of embryo production after intracytoplasmic sperm injection (ICSI) is still low because of frequent failure of normal fertilization, which involves formation of two polar bodies and two pronuclei. To clarify the reasons for this, we hypothesized that ICSI does not properly trigger sperm-induced fertilization events, especially intracellular Ca(2+) signaling, also known as Ca(2+) oscillation. We also suspected that the use of in vitro-matured oocytes might negatively affect fertilization events and embryonic development of sperm-injected oocytes...
September 30, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27725346/efficient-pig-icsi-using-percoll-selected-spermatozoa-evidence-for-the-essential-role-of-phospholipase-c-%C3%AE-in-icsi-success
#16
Michiko Nakai, Shun-Ichi Suzuki, Junya Ito, Dai-Ichiro Fuchimoto, Shoichiro Sembon, Junko Noguchi, Akira Onishi, Naomi Kashiwazaki, Kazuhiro Kikuchi
In pigs, the damaged sperm membrane leads to leakage of phospholipase C-ζ (PLCζ), which has been identified as a sperm factor, and a reduction of oocyte-activating ability. In this study, we investigated whether sperm selected by Percoll gradient centrifugation (Percoll) have sufficient PLCζ, and whether the efficiency of fertilization and blastocyst formation after intracytoplasmic sperm injection (ICSI) using Percoll-selected sperm can be improved. Percoll-selected sperm (Percoll group) or sperm without Percoll selection (Control group) were used...
September 30, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27665783/meiotic-cohesin-subunits-rad21l-and-rec8-are-positioned-at-distinct-regions-between-lateral-elements-and-transverse-filaments-in-the-synaptonemal-complex-of-mouse-spermatocytes
#17
Mei Rong, Atsushi Matsuda, Yasushi Hiraoka, Jibak Lee
Cohesins containing a meiosis-specific α-kleisin subunit, RAD21L or REC8, play roles in diverse aspects of meiotic chromosome dynamics including formation of axial elements (AEs), assembly of the synaptonemal complex (SC), recombination of homologous chromosomes, and cohesion of sister chromatids. However, the exact functions of individual α-kleisins remain to be elucidated. Here, we examined the localization of RAD21L and REC8 within the SC by super-resolution microscopy, 3D-SIM. We found that both RAD21L and REC8 were localized at the connection sites between lateral elements (LEs) and transverse filaments (TFs) of pachynema with RAD21L locating interior to REC8 sites...
September 26, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27647534/adenylation-by-testis-specific-cytoplasmic-poly-a-polymerase-papolb-tpap-is-essential-for-spermatogenesis
#18
Shin-Ichi Kashiwabara, Satsuki Tsuruta, Keitaro Okada, Yutaro Yamaoka, Tadashi Baba
The testis-specific cytoplasmic poly(A) polymerase PAPOLB/TPAP is essential for spermatogenesis. Although this enzyme is responsible for poly(A) tail extension of a subset of mRNAs in round spermatids, the stability and translational efficiency of these mRNAs are unaffected by the absence of PAPOLB. To clarify the functional importance of this enzyme's adenylation activity, we produced PAPOLB-null mice expressing a polyadenylation-defective PAPOLB mutant (PAPOLB(D114A)), in which the catalytic Asp at residue 114 was mutated to Ala...
September 18, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27616283/combination-of-density-gradient-centrifugation-and-swim-up-methods-effectively-decreases-morphologically-abnormal-sperms
#19
Masaya Yamanaka, Kazuhisa Tomita, Shu Hashimoto, Hiroshi Matsumoto, Manabu Satoh, Hiromi Kato, Yoshihiko Hosoi, Masayasu Inoue, Yoshiharu Nakaoka, Yoshiharu Morimoto
Density gradient centrifugation (DGC) and swim-up techniques have been reported for semen preparation in assisted reproductive techniques in humans. We investigated whether semen preparation using a combination of DGC and swim-up techniques could effectively decrease morphologically abnormal human sperms at the ultrastructural level. Semen samples were obtained from 16 infertile males and fractionated by swim-up following DGC. Ultrastructural abnormalities of sperms obtained from original semen, lower layer of swim-up following DGC, and upper layer of swim-up following DGC were analyzed by transmission electron microscopy...
September 11, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27301424/sex-sorting-of-spermatozoa-affects-developmental-competence-of-in-vitro-fertilized-oocytes-in-a-bull-dependent-manner
#20
Yasushi Inaba, Reika Abe, Masaya Geshi, Satoko Matoba, Takashi Nagai, Tamás Somfai
The aim of the present study was to clarify if flow-cytometric sex-sorting of bovine sperm affected in vitro blastocyst production in different bulls, either in terms of its ability to fertilize the oocyte or by interfering with post-fertilization embryo development. We performed in vitro fertilization (IVF) using both commercially available frozen-thawed X-sorted and non-sorted sperm of 4 Holstein bulls at 3 concentrations (1 × 10(6), 2 × 10(6), and 5 × 10(6) sperm/ml). When fertilization rates were compared, a variation in fertilization rates among different sperm concentrations was detected in 2 bulls, with similar results for X-sorted and non-sorted sperm...
October 18, 2016: Journal of Reproduction and Development
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