〈Original Papers〉卵細胞内精子注入法(ICSI)により作製した2細胞期胚の冷蔵輸送の検討

[Abstract] Previously, we have succeeded in transporting vitrified/warmed 2-cell mouse embryos while maintaining developmental ability at refrigerated temperatures for 50 hours. In this study, we examined the developmental ability of 2-cell stage embryos produced by Intracytoplasmic Sperm Injection (ICSI), which were then refrigerated and transported. Sampling tubes containing 2-cell stage embryos produced by ICSI were placed into embryo transport containers and transported from the Institute of Advanced Technology, Kinki University, Wakayama to the Center for Animal Resources and Development (CARD), Kumamoto University, Kumamoto. The time required for transport was around 2 days. During transport, the temperature of the tubes was kept between 2.5 and 5.0℃. At CARD, the embryos were recovered from the sampling tubes and transferred into the oviducts of pseudopregnant females on the day that a vaginal plug was found (Day 1 of pseudopregnancy). The rate of viable 2-cell stage embryos after transport was 100％ (122/122), of which 21％ developed to live young. [和文抄録] 超低温下(-196℃)における初期胚の凍結輸送に変わる手段として、より簡便な冷蔵下(4℃)における輸送方法について、高度な生殖補助技術を必要とする卵細胞内精子注入法(ICSI)により作成された初期胚を用いて検討した。C57BL/6Jを用いた卵細胞内精子注入法(ICSI)後の卵子は52％(205/392)が生存した。それらの受精率は73％(150/205)であり、その後の2細胞期胚への発生率は83％(124/150)であった。得られた2細胞期胚は少量のM2培地と共にテストチューブへ移し、冷蔵輸送は、クール宅急便にて近畿大学先端技術総合研究所(和歌山)から熊本大学・CARD(熊本)へ輸送した。輸送後の回収された生存胚は100％(122/122)であった。さらに偽妊娠を誘起したICR系マウスの卵管内に移植し発生能を検討したところ21％(26/122)が産子へ発生した。以上の結果より、ICSI由来胚の冷蔵輸送が可能であることを明らかにした

A ubiquitin ligase complex assembles linear polyubiquitin chains

The ubiquitin system plays important roles in the regulation of numerous cellular processes by conjugating ubiquitin to target proteins. In most cases, conjugation of polyubiquitin to target proteins regulates their function. In the polyubiquitin chains reported to date, ubiquitin monomers are linked via isopeptide bonds between an internal Lys and a C-terminal Gly. Here, we report that a protein complex consisting of two RING finger proteins, HOIL-1L and HOIP, exhibits ubiquitin polymerization activity by recognizing ubiquitin moieties of proteins. The polyubiquitin chain generated by the complex is not formed by Lys linkages, but by linkages between the C- and N-termini of ubiquitin, indicating that the ligase complex possesses a unique feature to assemble a novel head-to-tail linear polyubiquitin chain. Moreover, the complex regulates the stability of Ub-GFP (a GFP fusion protein with an N-terminal ubiquitin). The linear polyubiquitin chain generated post-translationally may function as a new modulator of proteins

Investigations of motility and fertilization potential in thawed cryopreserved mouse sperm from cold-stored epididymides

Cold transport of epididymides from genetically modified mice is an efficient alternative to the shipment of live animals between research facilities. Mouse sperm from epididymides cold-stored for short periods can maintain viability. We previously reported that cold storage of mouse epididymides in Lifor® perfusion medium prolonged sperm motility and fertilization potential and that the sperm efficiently fertilized oocytes when reduced glutathione was added to the fertilization medium. Cryopreservation usually results in decreased sperm viability; an optimized protocol for cold storage of epididymides plus sperm cryopreservation has yet to be established. Here, we examined the motility and fertilization potential of cryopreserved, thawed (frozen-thawed) sperm from previously cold-stored mouse epididymides. We also examined the protective effect of sphingosine-1-phosphate (S1P) on sperm viability when S1P was added to the preservation medium during cold storage. We assessed viability of frozen-thawed sperm from mouse epididymides that had been cold-transported domestically or internationally and investigated whether embryos fertilized in vitro with these sperm developed normally when implanted in pseudo-pregnant mice. Our results indicate that frozen-thawed sperm from epididymides cold-stored for up to 48 h maintained high fertilization potential. Fertilization potential was reduced after cold storage for 72 h, but not if S1P was included in the cold storage medium. Live pups were born normally to recipients after in vitro fertilization using frozen-thawed sperm from cold-transported epididymides. In summary, we demonstrate an improved protocol for cold-storage of epididymides that can facilitate transport of genetically engineered-mice and preserve sperm viability after cryopreservation

在宅痴呆性高齢者の住環境

When the family doing life together becomes dementia, confusion of family is thought that there isan immeasurable thing in the circumference. But, everyday life is no which it waited for, and it can\u27t always perplex it. A concrete countermeasure toward those conditions becomes necessary. However, how to catch it is vague, and serious dementia is being done as for the mild dementia , too. But, when the daily life of the dementia old man is observed firmly, it is understood that there is a difference in the way that a life difficulty appears. This time, life was done at home, and it the condition figure that it had the older adults with dementia using day care and dayservic seen at the life scene in five types. It is reported because a family, careworker, a occupational therapist etc did a discussion about the environment maintenance of the type with dementia and so on one rank respectively and a consideration was done