Zhao Namula, Takeshige Otoi, Theerawat Tharasanit, Kaywalee Chatdarong, Megumi Nagahara, Setyo Oky Widodo, Aya Nakai, Thi Suong Nguyen, Yuichiro Nakayama, Maki Hirata and Fuminori Tanihara : Establishment of a Single-Oocyte Culture System for Pigs and Its Validation Using Curcumin as a Model Antioxidant for Oocyte Maturation, Animals : An Open Access Journal from MDPI, 15, 22, 2025.
(Keyword)
curcumin / meiotic competence / microdroplet / porcine oocyte / single culture system
T. Suong Nguyen, Takeshige Otoi, Zhao Namula, Setyo Oky Widodo, Theerawat Tharasanit, Kaywalee Chatdarong, Yuichiro Nakayama, Megumi Nagahara, Aya Nakai, Maki Hirata and Fuminori Tanihara : Mitochondrial-Targeted Protective Potential of Elamipretide for the In Vitro Production of Porcine Embryos, Animals : An Open Access Journal from MDPI, 15, 17, 2497, 2025.
Bin Liu, Takeshige Otoi, Zhao Namula, Setyo Oky Widodo, Maki Hirata, Aya Nakai, Qingyi Lin, Yuichiro Nakayama, Megumi Nagahara and Fuminori Tanihara : Oxidative stress index of porcine follicular fluid influences meiotic maturation and embryo development during in vitro culture, Veterinary World, 18, 7, 2078-2086, 2025.
(Keyword)
embryo development / glutathione / in vitro maturation / oxidative stress index / porcine follicular fluid, reactive oxygen species
Aya Nakai, Shiho Fukushima, Kazuaki Mawatari, Akira Takahashi, Takeshi Nikawa, Shintaro Inoue, Takahito Watanabe, Taro Mito and Katsuyuki Miyawaki : Differential Expression of Key Isoflavone Synthesis Genes in Soybean Sprouts under Two LED Treatments, ACS Agricultural Science & Technology, 2025.
(Summary)
Soybean seeds and sprouts contain isoflavones, including aglycones, glucosides, and malonylated glucosides, and their contents vary across soybean varieties. During germination and photomorphogenesis, blue light-emitting diode (LED) irradiation suppressed hypocotyl growth and enhanced the expression levels of chalcone synthase 7, isoflavone synthase (IFS), and isoflavone reductase (IFR). Conversely, red LED irradiation enhanced hypocotyl growth and decreased the expression levels of IFS and IFR. The contents of some glycosides or malonylated glycosides in sprouts and young green soybeans were significantly higher under blue LED irradiation than under red LED irradiation. These results imply that blue light more effectively increased the concentrations of some isoflavones during early germination and/or hydroponics cultivation.
Nanaka Torigoe, Qingyi Lin, Bin Liu, Yuichiro Nakayama, Aya Nakai, Megumi Nagahara, Fuminori Tanihara, Maki Hirata and Takeshige Otoi : Effects of Electroporation Timing and Cumulus Cell Attachment on In Vitro Development and Genome Editing of Porcine Embryos., Reproduction in Domestic Animals = Zuchthygiene, 60, 2, e70011, 2025.
(Summary)
Pig genome editing using the oviductal nucleic acid delivery (GONAD) method with electroporation would allow the efficient obtention of genetically modified pigs. However, oocytes and zygotes at various stages after ovulation must be targeted, and cumulus cell attachment and mosaic mutations are major obstacles. Therefore, we investigated whether two parameters (electroporation timing and the cumulus cell attachment) influence the effectiveness of multiplex genome editing by electroporation in porcine oocytes or zygotes. Three gRNAs targeting either GGTA1, CMAH or B4GALNT2 were introduced individually into oocytes and zygotes with and without cumulus cells at three different time points, 0 h before in vitro fertilisation (IVF) and 5 h and 10 h after IVF initiation. The introduction of gRNAs into oocytes and zygotes did not significantly affect the rates of blastocyst formation and total mutation of the resulting blastocysts irrespective of cumulus cell attachment and electroporation timing. In conclusion, the electroporation timing and the cumulus cell attachment did not interfere with the efficient delivery of the CRISPR/Cas9 system to the oocytes/zygotes, indicating that porcine genome editing in the oviduct using GONAD method may be possible.
(Keyword)
Animals / Cumulus Cells / Electroporation / Gene Editing / Female / Oocytes / CRISPR-Cas Systems / Fertilization in Vitro / Embryonic Development / Swine / Zygote / RNA, Guide, CRISPR-Cas Systems / Blastocyst / Galactosyltransferases / Embryo Culture Techniques
Supitcha Kaewma, Takeshige Otoi, Setyo Oky Widodo, Megumi Nagahara, Aya Nakai, T Suong Nguyen, Yuichiro Nakayama, Theerawat Tharasanit, Maki Hirata and Fuminori Tanihara : Zinc Chloride Supplementation During In Vitro Fertilization Reduces Polyspermic Fertilization of Porcine Oocytes, Animal Science Journal, 96, 1, e70095, 2025.
(Summary)
As zinc is important for the fertilization competency of sperms and oocytes, we investigated the effects of zinc chloride (ZnCl2) supplementation during in vitro fertilization (IVF) on porcine oocyte fertilization and development. We evaluated the effects of ZnCl2 concentration (0, 1, 10, and 20 μg/mL) on the quality of frozen-thawed boar spermatozoa cultured for 5 h and on oocyte fertilization and embryo development after IVF. Spermatozoa from three different boars were additionally tested. ZnCl2 supplementation effects along with metal chelators, calcium ethylenediaminetetraacetic acid (Ca-EDTA) and zinc EDTA (Zn-EDTA), were also examined. ZnCl2 supplementation did not affect the quality of frozen-thawed spermatozoa after culture for 5 h. Supplementation with 1-μg/mL ZnCl2 decreased the percentage of polyspermic fertilization compared to that at 0- and 20-μg/mL ZnCl2. Moreover, it increased blastocyst formation rate compared to that in the other supplementation groups. In different boar spermatozoa, ZnCl2 supplementation (1 μg/mL) decreased polyspermic fertilization but did not improve embryo development. Co-incubation with Ca-EDTA did not reduce polyspermic fertilization, but Zn-EDTA co-incubation reduced polyspermic fertilization similar to that with ZnCl2 alone. In conclusion, 1-μg/mL ZnCl2 supplementation during IVF reduces polyspermic fertilization but may not improve embryo development.
(Keyword)
embryonal development / in vitro fertilization / monospermy / pig / zinc
Qingyi Lin, Nanaka Torigoe, Bin Liu, Yuichiro Nakayama, Aya Nakai, Zhao Namula, Megumi Nagahara, Fuminori Tanihara, Maki Hirata and Takeshige Otoi : Efficient gene editing of pig embryos by combining electroporation and lipofection., Veterinary World, 17, 11, 2701-2707, 2024.
(Summary)
Electroporation combined with lipofection using gRNAs with low mutation rates may improve gene editing efficiency in pig embryos. However, the effects may vary based on the timing of gene editing.
Koki Takebayashi, Manita Wittayarat, Maki Hirata, Qingyi Lin, Zhao Namula, Nanaka Torigoe, Bin Liu, Megumi Nagahara, Aya Nakai, Takeshige Otoi and Fuminori Tanihara : Optimization of embryonic stage for aggregation to generate chimeric pigs using gene-edited blastomeres., In Vitro Cellular & Developmental Biology. Animal, 2024.
Aya Nakai, Akihito Tanaka, Hitoshi Yoshihara, Koji Murai, Takahito Watanabe and Katsuyuki Miyawaki : Blue LED light promotes indican accumulation and flowering in indigo plant, Polygonum tinctorium, Industrial Crops and Products, 155, 112774, 2020.
(Keyword)
Polygonum tinctorium / Plant factory / Blue LED / Photoperiod / Flowering / Indican