Reproductive Immunology and Biology
Online ISSN : 1881-7211
Print ISSN : 1881-607X
ISSN-L : 1881-607X
Volume 22, Issue 1
Displaying 1-2 of 2 articles from this issue
  • Akiko HASEGAWA, Nozomi KANAZAWA, Koji KOYAMA
    2007 Volume 22 Issue 1 Pages 1-7
    Published: 2007
    Released on J-STAGE: September 01, 2009
    JOURNAL FREE ACCESS
    The zona pellucida (ZP) surrounding the mammalian oocyte plays an important roles during fertilization and pre-implantation embryo development. In this study, we examined whether the mouse ZP components of ZP1, ZP2 and ZP3 are involved in follicular development using RNA interference (RNAi) techniques. Premature follicles were collected from juvenile mouse ovaries and short-interfering RNAs (siRNAs) were injected into the oocytes through the granulosa cell layers. After 6-day culture, oocyte survival rates and diameters of surviving oocytes were measured. The effect of siRNA was confirmed by RT-PCR from single oocytes and immunofluorescent staining with specific antibodies. Inhibition of ZP1, ZP2 and ZP3 synthesis by the corresponding siRNAs resulted in the impairment of oocyte growth and follicular development as well as ZP formation. Among the three ZP proteins, ZP3 showed the strongest inhibition in this experiment. These results suggested that all the ZP proteins and particularly ZP3 were associated with oocyte growth and follicular development in the ovary.
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  • Yorino SATO, Kahei SATO
    2007 Volume 22 Issue 1 Pages 8-13
    Published: 2007
    Released on J-STAGE: September 01, 2009
    JOURNAL FREE ACCESS
    Recently, the spermatogenic cell has been focused in male sterility. There are some reports on fertilization and embryonic developmental ability of spermatid. Deficiency of fertilization or developmental arrest frequently appears due to the ability. Functional immaturity is also observed in spite of abnormality of karyotype in spermatids. However, a functional difference between spermatids and spermatozoa is not clarified. In this study, fertilization and developmental ability of the spermatid were examined by the microinjection using immature spermatid including the round spermatid, elongating spermatid, elongated spermatid and intratesticular spermatozoa derived from three different mice strains; ddY, ICR and B6D2F1. Mature female mice were superovulated with PMSG and hCG. Cumulus-oosytes complex were obtained from their oviducts, and cumulus cells were removed from MII oocytes by hyaluronidase and pippeting. Spermatids were collected from mature male testis, and were injected into MII oocytes using a micromanipulator. Manipulated embryos were cultured in CZB medium, and their ability for development was examined. Epididymal spermatozoa from the three strains were used as control group. The fertilization rate was correlated with the spermatogenic stage. In embryonic developmental rate, on the other hand, there were no significant difference between the control and experimental embryos. The data however showed that elongating, elongated and round spermatid slightly deteriorated the embryonic development. These results suggest that the ability of the embryonic activation depend on the spermatid metamorphose.
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