Yasushi Takai

Preimplantation exposure to bisphenol A advances postnatal development

Prenatal exposure to bisphenol A (BPA), an estrogenic compound, has been shown to alter postnatal development at an environmentally relevant exposure level. To elucidate these low dose effects of preimplatation exposure to BPA, two-cell mouse embryos were cultured with 1 nM BPA. More embryos exposed to 1 nM BPA than controls reached the blastocyst stage. When the blastocysts with or without BPA exposure were transferred to uterine horns of pseudopregnant recipient mice not treated with BPA, the number of pups per litter and body weight at birth did not differ. At weaning on postnatal day 21, however, pups treated with 1 nM BPA during the preimplantation period were significantly heavier than controls. These findings suggest that BPA may not only affect early embryonic development even at low, environmentally relevant doses, but also may exert late effects on postnatal development.

A central role for ceramide in the age-related acceleration of apoptosis in the female germline

An age‐dependent acceleration of apoptosis occurs in female germ cells (oocytes), and this requires communication between the oocyte and its surrounding somatic (cumulus) cells. Here we show in aged mice that ceramide is translocated from cumulus cells into the adjacent oocyte and induces germ cell apoptosis that can be prevented by sphingosine‐1‐phosphate. Trafficking of ceramide requires gap junction‐dependent communication between the cumulus cells and the oocyte as well as intact lipid rafts. Further, the occurrence of the elevated incidence of apoptosis in oocytes of aged females is concomitant with an enhanced sensitivity of the oocyte to a spike in cytosolic ceramide levels, as well as increased bax mRNA and Bax protein levels. Thus, the force driving the age‐related increase in female germ cell death is multifactorial, but changes in the intercellular trafficking of ceramide, along with hypersensitivity of oocytes to ceramide, are key factors in this process.

Phosphorylation by Cdk1 induces Plk1-mediated vimentin phosphorylation during mitosis

Several kinases phosphorylate vimentin, the most common intermediate filament protein, in mitosis. Aurora-B and Rho-kinase regulate vimentin filament separation through the cleavage furrow-specific vimentin phosphorylation. Cdk1 also phosphorylates vimentin from prometaphase to metaphase, but its significance has remained unknown. Here we demonstrated a direct interaction between Plk1 and vimentin-Ser55 phosphorylated by Cdk1, an event that led to Plk1 activation and further vimentin phosphorylation. Plk1 phosphorylated vimentin at ∼1 mol phosphate/mol substrate, which partly inhibited its filament forming ability, in vitro. Plk1 induced the phosphorylation of vimentin-Ser82, which was elevated from metaphase and maintained until the end of mitosis. This elevation followed the Cdk1-induced vimentin-Ser55 phosphorylation, and was impaired by Plk1 depletion. Mutational analyses revealed that Plk1-induced vimentin-Ser82 phosphorylation plays an important role in vimentin filaments segregation, coordinately with Rho-kinase and Aurora-B. Taken together, these results indicated a novel mechanism that Cdk1 regulated mitotic vimentin phosphorylation via not only a direct enzyme reaction but also Plk1 recruitment to vimentin.

Absence of the proapoptotic Bax protein extends fertility and alleviates age-related health complications in female mice

The menopausal transition in human females, which is driven by a loss of cyclic ovarian function, occurs around age 50 and is thought to underlie the emergence of an array of health problems in aging women. Although mice do not undergo a true menopause, female mice exhibit ovarian failure long before death because of chronological age and subsequently develop many of the same age-associated health complications observed in postmenopausal women. Here we show in mice that inactivation of the proapoptotic Bax gene, which sustains ovarian lifespan into advanced age, extends fertile potential and minimizes many age-related health problems, including bone and muscle loss, excess fat deposition, alopecia, cataracts, deafness, increased anxiety, and selective attention deficit. Further, ovariectomy studies show that the health benefits gained by aged females from Bax deficiency reflect a complex interplay between ovary-dependent and -independent pathways. Importantly, and contrary to popular belief, prolongation of ovarian function into advanced age by Bax deficiency did not lead to an increase in tumor incidence. Thus, the development of methods for postponing ovarian failure at menopause may represent an attractive option for improving the quality of life in aging females.

Stem cell factor (SCF) concentrations in peritoneal fluid of women with or without endometriosis.

PROBLEM: In the quest for possible involvement of stem cell factor (SCF), a cytokine known to have multiple effects, in the pathogenesis of endometriosis, we evaluated concentrations of SCF in peritoneal fluid (PF) of women with or without endometriosis.
 METHOD OF STUDY: SCF concentrations in PF collected from women undergoing laparoscopy were measured, using a specific enzyme‐linked immunosorbent assay (ELISA). Reverse transcription‐polymerase chain reaction (RT‐PCR) analysis to detect gene expression of c‐kit, the receptor for SCF, was performed using the endometriotic tissue and the eutopic endometrium collected during the operation.
 RESULTS: SCF concentrations in PF of women with endometriosis were significantly higher compared to women without endometriosis. Looking at SCF concentrations in PF of women with endometriosis stratified by disease stage, women with stage I and II exhibited relatively higher SCF levels in PF, whereas SCF levels in PF with stage III and IV were comparable with those without endometriosis. The expression of mRNA for c‐kit was detected in both the endometriotic tissue and the eutopic endometrium.
 CONCLUSION: We demonstrated an elevation in SCF levels in PF associated with endometriosis and the presence of its receptor in endometriotic tissues. Given the known pleiotropic properties of SCF, the present results suggest that SCF might play a role in the pathogenesis of endometriosis.

Caspase-12 compensates for lack of caspase-2 and caspase-3 in female germ cells

Previously, we analyzed mice lacking either caspase-2 or caspase-3 and documented a role for caspase-2 in developmental and chemotherapy-induced apoptosis of oocytes. Those data also revealed dispensability of caspase-3, although we found this caspase critical for ovarian granulosa cell death. Because of the mutual interdependence of germ cells and granulosa cells, herein we generated caspase-2 and -3 double-mutant (DKO) mice to evaluate how these two caspases functionally relate to each other in orchestrating oocyte apoptosis. No difference was observed in the rate of spontaneous oocyte apoptosis between DKO and wildtype (WT) females. In contrast, the oocytes from DKO females were more susceptible to apoptosis induced by DNA damaging agents, compared with oocytes from WT females. This increased sensitivity to death of DKO oocytes appears to be a specific response to DNA damage, and it was associated with a compensatory upregulation of caspase-12. Interestingly, DKO oocytes were more resistant to apoptosis induced by methotrexate (MTX) than WT oocytes. These results revealed that in female germ cells, insults that directly interfere with their metabolic status (e.g. MTX) require caspase-2 and caspase-3 as obligatory executioners of the ensuing cell death cascade. However, when DNA damage is involved, and in the absence of caspase-2 and -3, caspase-12 becomes upregulated and mediates apoptosis in oocytes.

Rapid Detection of Chromosome Aneuploidies by Prenatal Interphase FISH (Fluorescence in situ Hybridization) and Its Clinical Utility in Japan

Objective: The purpose of this study was to assess the accuracy, informative rate, detection rate, and clinical utility of prenatal interphase fluorescence in situ hybridization (FISH) analysis of amniotic fluid samples from Japanese women.

Breast-fed infants, possibly exposed to dioxins in milk, have unexpectedly lower incidence of endometriosis in adult life

Ž . 2,3,7,8-Tetrachlorodibenzo-p-dioxin TCDD is the most toxic congener among polychlorinated dibenzodioxins and polychlorinated dibenzofurans. These compounds are resistant to degradation and are ubiquitous environmental pollutants that become concentrated in animal tissues, climb the food chain, accumulate in adipose tissue and are excreted in breast milk in large amounts. The estimated daily dietary intake of these pollutants by breast-fed infants was reported to amount to 170 pgrkg per day at 2 months of age which is higher than the tolerable daily intake recomŽ mended by the WHO-10 most recently, 1]4 . pgrkg per day . In Japan, like other industrialized countries, dioxin contamination is a serious public problem. Recent government reports indicated that daily dioxin intake of average Japanese is

Quantitative assessment of human leukocyte antigen-G protein in amniotic fluid by a double-determinant enzyme-linked immunosorbent assay using anti-human leukocyte antigen-G-specific antibody '87G'.

PROBLEM: The objective of this study was to establish an enzyme‐linked immunosorbent assay (ELISA) system, in an attempt to quantify the amount of human leukocyte antigen (HLA)‐G protein in amniotic fluid.

Dimeric Inhibin A as a Fourth Marker for Down's Syndrome Maternal Serum Screening in Native Japanese Women

Objective: This study was conducted to assess the usefulness of dimeric inhibin A as a fourth marker for Downs syndrome screening in addition to AFP, hCG and uE3 markers for native Japanese women.