Evaluation of maturation competence of metaphase II oocytes in mice based on the distance between pericentriolar materials of meiotic spindle: distance of PCM during oocyte maturation.

Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
Journal of Assisted Reproduction and Genetics (Impact Factor: 1.82). 11/2010; 28(2):157-66. DOI: 10.1007/s10815-010-9496-2
Source: PubMed

ABSTRACT To ascertain whether metaphase II (MII) spindle shape influences oocyte competence, we examined the meiotic spindle organization in in vivo ovulated (IVO) oocytes and in spontaneously matured or follicle stimulating hormone (FSH)-induced oocytes.
FSH-induced oocytes matured in Waymouth's MB752/1 or human tubal fluid (HTF) media and oocytes matured spontaneously in the basal medium were obtained, and spindles were detected by immunofluorescence. To evaluate the fertilization-associated differences in spindle morphology, we performed in vitro fertilization and analysed integrin mRNA expression.
The distance between the pericentriolar materials (PCMs) in oocytes matured under all conditions was initially more, but it reduced gradually and increased again thereafter. Therefore, oocytes exhibiting a reduction in the distance between PCMs had the highest development rate to blastocyst in each condition.
These results indicate that the 'maturation competence' of MII oocytes can be evaluated on the basis of the distance between PCMs.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: 1. A definite chronological sequence of events occurs in the eggs and follicles of rabbits after mating or after the injection of ovulation-inducing substances. The follicle secretes secondary liquor folliculi, and there occurs a separation of the corona radiata from strands connecting it to the follicle cells. The ovum goes through nuclear maturation with as climax the production of the first polar body by the 8th hour after copulation. 2. Thyroxin injections cause indirectly the same effects as mating or pituitary injections but no ovulation occurs. The thyroxin effect occurs later than the pituitary effect and is due to an initiation of atresia in the follicles. 3. Explantation of ova results in typical maturation phenomena which are apparently unaffected by the presence of pituitary hormones or of thyroxin in the culture medium. 4. It is concluded that maturation of the ovum can be obtained simply by isolating it from the normal follicular environment. 5. Normal fertilization can be secured with eggs removed from the follicles.
    Journal of Experimental Medicine 10/1935; 62(5):665-675. · 13.91 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The possible mediatory role of cAMP in the induction of oocyte maturation by luteinizing hormone (LH) is not yet clear since evidence for both inhibitory and stimulatory actions of the nucleotide on the oocyte has been provided. To elucidate the role of cAMP in regulation of oocyte meiosis we tried in the present study to dissociate between the inhibitory and stimulatory action of this nucleotide on oocyte maturation. To induce maturation, oocytes enclosed by their follicles were transiently exposed to either dibutyryl cAMP (dbcAMP) or to the phosphodiesterase inhibitor methylisobutylxanthine (MIX). Inhibition of maturation was obtained by the addition of the above agents to either follicle-enclosed oocytes incubated in the presence of LH or isolated cumulus-free oocytes that mature spontaneously in vitro. We found that inhibition of oocyte maturation is obtained by a relatively low dose of either dbcAMP or MIX while higher concentrations of these agents are required to induce oocyte maturation. Coupling of the oocyte to the cumulus cells, as expressed by the fraction of labeled uridine transferred from the cumulus cells to the oocyte following exposure of the follicle-enclosed cumulus-oocyte complex to MIX, was also determined. We found that uncoupling of the oocyte from the cumulus cells corresponded with the induction, but not inhibition of oocyte maturation, both by its concentration dependence and time-course. We suggest that cAMP has a dual role in regulation of oocyte maturation. Lower levels of the nucleotide act to maintain meiotic arrest, while elevated levels of cAMP mediate LH action to induce meiosis resumption.
    Molecular and Cellular Endocrinology 04/1988; 56(1-2):115-21. · 4.24 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The intracellular levels of cAMP play a critical role in the meiotic arrest of mammalian oocytes. However, it is debated whether this second messenger is produced endogenously by the oocytes or is maintained at levels inhibitory to meiotic resumption via diffusion from somatic cells. Here, we demonstrate that adenylyl cyclase genes and corresponding proteins are expressed in rodent oocytes. The mRNA coding for the AC3 isoform of adenylyl cyclase was detected in rat and mouse oocytes by RT-PCR and by in situ hybridization. The expression of AC3 protein was confirmed by immunocytochemistry and immunofluorescence analysis in oocytes in situ. Cyclic AMP accumulation in denuded oocytes was increased by incubation with forskolin, and this stimulation was abolished by increasing intraoocyte Ca(2+) with the ionophore A23187. The Ca(2+) effects were reversed by an inhibitor of Ca(2+), calmodulin-dependent kinase II. These regulations of cAMP levels indicate that the major cyclase that produces cAMP in the rat oocyte has properties identical to those of recombinant or endogenous AC3 expressed in somatic cells. Furthermore, mouse oocytes deficient in AC3 show signs of a defect in meiotic arrest in vivo and accelerated spontaneous maturation in vitro. Collectively, these data provide evidence that an adenylyl cyclase is functional in rodent oocytes and that its activity is involved in the control of oocyte meiotic arrest.
    Developmental Biology 07/2003; 258(2):385-96. · 3.64 Impact Factor


Available from