Biomedical subjects

J Ten

Publications and source records attributed to J Ten.

6 recordsLinked to original sources

PubMed1999

Effect of intracellular Ca2+ chelation with the acetoxymethyl ester-derived form of bis(o-aminophenoxy)ethane-N,N,N,N',N'-tetraacetic acid on meiotic division and chromosomal segregation in mouse oocytes.

PURPOSE: Our purpose was to ascertain the effect of intracellular Ca2+ chelation on the chromosomal distribution and segregation of mouse oocytes during maturation in vitro. METHODS: Germinal vesicle oocytes were loaded with the acetoxymethyl ester-derived form of bis(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM). Chromosomal distribution and segregation of control and BAPTA-AM-treated metaphase II (MII) oocytes were evaluated at 16 hr, and intracellular ATP content at 0, 1, and 16 hr after BAPTA-AM loading. RESULTS: BAPTA-AM treatment decreased (P < or = 0.05) the potential for in vitro maturation, increased (P < or = 0.0001) the percentage of oocytes displaying an abnormal distribution of metaphase II chromosomes in the meiosis II spindle and aneuploidy, and decreased (P < or = 0.005) the ATP content at 0, 1, and 16 hr of culture compared to the control groups. CONCLUSIONS: These findings raise some concern about any other condition/drug that may directly or indirectly decrease the intracellular Ca2+ concentration in human oocytes.

Adenosine Triphosphate↗

PubMed1998

Dithiothreitol prevents age-associated decrease in oocyte/conceptus viability in vitro.

The present study was designed to ascertain whether the negative effects on reproductive potential of post-ovulatory ageing in vitro of oocytes can be prevented by antioxidant therapy. Mouse metaphase II (MII) oocytes were aged in vitro for 12 h prior to insemination in the presence of varying concentrations of L-ascorbic acid, 6-methoxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox), L-cystine dihydrochloride, ethylenediaminetetraacetic acid (EDTA), beta-mercaptoethanol and DL-dithiothreitol (DTT). In-vitro ageing of oocytes was associated with lower fertilization rate, higher proportion of concepti exhibiting cellular fragmentation at 24 h post-insemination and lower percentage of concepti reaching the blastocyst stage. Ascorbic acid, Trolox and EDTA had no effect on cellular fragmentation or potential of oocytes for development. However, the probability of an oocyte reaching the blastocyst stage was decreased (P < or = or = 0.05) in oocytes incubated in the presence of L-cystine (50 and 500 microM) and beta-mercaptoethanol (5, 50 and 500 microM) when compared to control aged oocytes. Age-associated cellular fragmentation at 24 h post-insemination was partially prevented (P < or = 0.05) by incubating oocytes in the presence of beta-mercaptoethanol (500 microM). DTT (50 and 500 microM) increased (P < or = 0.05) fertilization rate and number of cells at 81 h post-insemination to levels similar to those exhibited by control oocytes. Furthermore, both age-associated fragmentation at 24 h post-insemination (P < or = 0.05) and decreased potential of oocytes for development to the blastocyst stage (P < or = 0.05) were prevented, at least in part, by culturing oocytes in the presence of DTT (50 microM). Although the mechanism by which DTT exerts its beneficial effects on aged oocytes remains to be elucidated, it may protect oocytes by preventing oxidation of free thiol groups and/or altering a redox-independent signalling pathway that mediates cellular fragmentation and death.

Animals↗

PubMed1998

Antioxidant therapy counteracts the disturbing effects of diamide and maternal ageing on meiotic division and chromosomal segregation in mouse oocytes.

This study aims (i) to ascertain whether oxidative-stress-induced disturbances in chromosomal distribution in the metaphase-II spindle of mouse oocytes can be counteracted by supplementing culture medium with antioxidants; and (ii) to determine whether supplemental intake of antioxidants neutralizes the disturbing effects of maternal ageing on segregation of chromosomes during the first meiotic division and distribution of chromosomes in the metaphase-II spindle. (i): Germinal vesicle oocytes from unstimulated 10-12 week old mice were matured in vitro in the presence or absence of diamide and/or dithiothreitol. Metaphase-II oocytes were fixed and stained with 4',6-diamidino-2-phenylindole (DAPI) to detect abnormalities in chromosomal distribution. The percentage of oocytes arrested in metaphase I (12.9% vs 28.4%; P < or = 0.05) or with a telophase-I chromosome configuration (0.0% vs 8.2%; P < or = 0.0005) was decreased in diamide-DTT-treated oocytes when compared to diamide-treated oocytes. (ii): Mice were fed, from the first day of weaning until their death, a diet supplemented or not with an antioxidant mixture of vitamin C and vitamin E. Ovulated oocytes were fixed and stained with DAPI or C-banded for chromosome analysis. The percentage of abnormal (chromosome scattering and nulloploidy) or asynchronous (anaphase I or telophase I) oocytes was 2.7-fold higher in controls than in females fed an antioxidant diet (24.4% vs 8.9%, P < or = 0.05). Furthermore, the percentage of aneuploidy (2.2% vs 0.0%; P < or = 0.01) and diploidy (5.8% vs 1.7%; P < or = 0.05) was significantly higher in controls than in females fed an antioxidant diet. These findings support Tarin's oxidative stress hypothesis of aneuploidy and have clinical implications for preventing both laboratory-induced and maternal-age-associated aneuploidy in human beings.

Aging↗

PubMed1996

The oxidizing agent tertiary butyl hydroperoxide induces disturbances in spindle organization, c-meiosis, and aneuploidy in mouse oocytes.

It has been recently proposed that a concomitant generation of oxidative stress of oocytes with increasing maternal age may be a major factor responsible for the age-related increase in aneuploid conceptions. As a preliminary step in the testing of this hypothesis, we need to confirm that oxidative stress in itself can induce errors in chromosome segregation. In order to achieve this goal, germinal vesicle (GV)-stage mouse oocytes from unstimulated ICR and (C57BL x CBA) F1 hybrid female mice were matured in vitro for 9 h for metaphase I (MI) oocytes or 16 h for metaphase II (MII) oocytes in the presence of varying concentrations of the oxidizing agent tertiary-butyl hydroperoxide (tBH). MII oocytes from (C57BL x CBA) F1 hybrid mice were fixed and C-banded for karyotyping analysis. MI and MII oocytes from ICR mice were fixed and stained with the DNA-fluorescent probe 4',6-diamidino-2-phenylindole (DAPI) to detect abnormalities in chromosomal distribution. Meiosis I and meiosis II spindles from ICR mice were visualized by confocal immunofluorescence microscopy. Data from these experiments demonstrate that in-vitro exposure of mouse oocytes to tBH during meiosis I reduces the length (pole-to-pole distance) and width (diameter at the equator of the spindle) of meiosis I and meiosis II spindles. This reduction is associated with an increase in the percentage of oocytes showing chromosome scattering and clumping on the MII plate, and of aneuploidy (hyperhaploidy) in MII oocytes. However, tBH at the concentrations used in the present study has only a minimal negative effect on the frequency of meiotic maturation. These results suggest that oxidative stress during meiotic maturation in vitro may induce chromosomal errors that are undetectable in the living oocyte and whose developmental consequences may become manifest after fertilization.

Aneuploidy↗

PubMedDate not supplied

Dietary antioxidant supplementation did not affect declining sperm function with age in the mouse but did increase head abnormalities and reduced sperm production.

The present study aims to ascertain whether dietary administration of a mixture of vitamins C and E may prevent the negative effects of paternal age on male fertility variables in the mouse. Experimental males were fed a standard diet supplemented with either a low or high dose of vitamins C and E. Oocytes enclosed in cumulus masses were inseminated using a 2 x 2 factorial design established according to whether males were young (3-4 months of age) or aged (22-24 months of age) and whether they were fed a control or antioxidant diet. Aged males showed a significant decrease in number of spermatozoa/mg epididymis when compared to young males. Dietary supplementation with low doses of vitamins C and E did not have any effect on sperm quality, fertilization and conceptus development in vitro. However, high doses of dietary vitamins C and E decreased the number of spermatozoa/mg epididymis, and increased the percentage of spermatozoa with misshapen heads distended in the distal part. These findings suggest that dietary supplementation with pharmacological doses of vitamins C and E may disturb spermatogenesis. The mechanism involved may be associated with either the antioxidant properties of vitamins C and E or their inhibitory action on steroidogenesis by Leydig and/or Sertoli cells.

Aging↗

PubMedDate not supplied

Effects of maternal ageing and dietary antioxidant supplementation on ovulation, fertilisation and embryo development in vitro in the mouse.

The present study aims to ascertain whether dietary supplementation with a mixture of vitamins C and E may prevent the maternal-age-associated decrease in both the number of ovulated oocytes after exogenous ovarian stimulation and embryo development in vitro in the mouse. Experimental females were fed a standard diet supplemented with i) high doses of vitamins C and E from the first day of weaning until 12 or 40 weeks of age; or ii) moderate doses of vitamins C and E from the first day of weaning until 12 weeks of age or from 22 to 33 weeks of age. The age-related reduction in ovulation rate was partially prevented by supplementing diet with high doses of vitamins C and E from the first day of weaning. Shorter periods of treatment and lower doses of vitamins C and E were also efficient in preventing the maternal-age-associated reduction in ovulation rate after exogenous ovarian stimulation. No effect of maternal diet on fertilisation and embryo development was observed until the blastocyst stage. Although any extrapolation to human fertility should be made with caution, these findings may have direct implications for preventing or delaying maternal-age-associated infertility in humans.

Aging↗