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PubMed1985

CA++ in fertilization and mitosis: the phosphatidylinositol cycle in sea urchin gametes and zygotes is involved in control of fertilization and mitosis.

We determined that the phosphatidylinositol (PI) cycles in both sea urchin sperm and eggs are necessary for normal fertilization, and that the PI cycle in sea urchin zygotes is involved in control of mitosis. The PI cycle is involved in Ca++ homeostasis so our data are direct evidence that Ca++ is involved with control of mitosis and fertilization. We implicated the PI cycle by adding Li+ to sea urchin eggs, sperm, or zygotes: those effects of Li+ due to effects on the PI cycle were overcome by myo-inositol but not by its optical isomer, scyllitol, and not by mannitol.

Animals↗
PubMed1998

Cleavage, development and competence of sheep embryos fertilized by intracytoplasmic sperm injection and in vitro fertilization.

More abnormal fertilization has been found in sheep oocytes after intracytoplasmic sperm injection (ICSI) than after in vitro fertilization (IVF). Although the birth of a normal lamb has been reported, the efficiency of blastocyst production is low. We therefore evaluated the cleavage, development and viability of sheep embryos obtained from ICSI, IVF and sham injection. In vitro matured oocytes either injected or inseminated with spermatozoa were assessed for cleavage 1 and 4 d after injection or insemination, and for development to blastocyst after 7 d of culture. A total of 699 oocytes was injected (ICSI); 198 (30.6%) were activated and 55 (8.5%) developed to the blastocyst stage. Of the 17 recipient ewes with 1, 2, 3 or 4 embryos, 15 (88.2%) were pregnant on Day 18; of these 17 recipients, 7 (41.1%) and 6 (35.2%) ewes remained pregnant on Days 45 and 110, respectively. Two normal lambs were born, one ewe died on Day 110 with 2 normal male fetuses, another ewe aborted on Day 90 and 4 pregnancies were maintained. A total of 517 oocytes was inseminated (IVF); 296 (62%) were activated and 90 (18.8%) reached the blastocyst stage. A total of 19 ewes received 1, 2, 3 or 4 embryos; of these, 13 (68.4%) were pregnant on Day 18, 8 (42.1%) ewes remained pregnant on each of Days 45 and 110. Three ewes delivered 5 lambs. Five pregnancies were maintained. A total of 156 oocytes was sham injected, 38 (24.3%) were activated and no blatocysts were obtained after culture. The results of this study showed that blastocysts obtained after ICSI are potentially viable and are not a result of parthenogenesis.

Animals↗
PubMed1995

Coculture of ovine zygotes fertilized in vivo or in vitro and positive effect of CZB medium on the development of in vitro fertilized zygotes.

The development and quality of ovine zygotes derived from in vivo (IVOF) or in vitro fertilization (IVF) were compared after coculture on sheep oviductal cells. The same criteria were used to evaluate the coculture of IVF zygotes in CZB medium for 2 d followed by 199 medium for 6 d (CZB-199 coculture) or in 199 medium for 8 d (199 coculture). A higher overall developmental rate to blastocyst stages was obtained with IVOF (65.7%) than with IVF (23.2%) zygotes. More IVF zygotes reached blastocyst stages in CZB-199 (36.7%) than in 199 coculture (22.9%). The morphological aspect did not differ significantly between IVOF and IVF or between 199 and CZB-199 blastocysts. Histological examination revealed no significant difference in the pyknotic and mitotic indices and mean number of cells in the trophoblast and in the inner cell mass of hatched blastocysts between IVOF and IVF or between CZB-199 and 199 cocultures. According to criteria used in this study, the quality of blastocysts was equivalent, independently of fertilization or coculture systems. The use of CZB medium during the first cleavages increases the proportion of blastocysts.

Animals↗
PubMed1985

Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization.

Microtubules forming within the mouse egg during fertilization are required for the movements leading to the union of the sperm and egg nuclei (male and female pronuclei, respectively). In the unfertilized oocyte, microtubules are predominantly found in the arrested meiotic spindle. At the time for sperm incorporation, a dozen cytoplasmic asters assemble, often associated with the pronuclei. As the pronuclei move to the egg center, these asters enlarge into a dense array. At the end of first interphase, the dense array disassembles and is replaced by sheaths of microtubules surrounding the adjacent pronuclei. Syngamy (pronuclear fusion) is not observed; rather the adjacent paternal and maternal chromosome sets first meet at metaphase. The mitotic apparatus emerges from these perinuclear microtubules and is barrel-shaped and anastral, reminiscent of plant cell spindles; the sperm centriole does not nucleate mitotic microtubules. After cleavage, monasters extend from each blastomere nucleus. The second division mitotic spindles also have broad poles, though by third and later divisions the spindles are typical for higher animals, with narrow mitotic poles and fusiform shapes. Colcemid, griseofulvin, and nocodazole inhibit the microtubule formation and prevent the movements leading to pronuclear union; the meiotic spindle is disassembled, and the maternal chromosomes are scattered throughout the oocyte cortex. These results indicate that microtubules forming within fertilized mouse oocytes are required for the union of the sperm and egg nuclei and raise questions about the paternal inheritance of centrioles in mammals.

Animals↗
PubMed1995

A precise structural analysis of a fertilization-associated carbohydrate-rich glycopeptide isolated from the fertilized eggs of euryhaline killi fish (Fundulus heteroclitus). Novel penta-antennary N-glycan chains with a bisecting N-acetylglucosaminyl residue.

A novel carbohydrate-rich sialoglycopeptide of apparent molecular mass approximately 6 kDa was isolated from the fertilized eggs of Fundulus heteroclitus (euryhaline killi fish). This glycopeptide is a member of the L-hyosophorin family, characterized by its high content of carbohydrate (80-90% by weight) and formed by depolymerization of the precursor glycopoly-protein (H-hyosophorin) upon fertilization. The structures of the N-glycan chains were unambiguously established by a combination of compositional analysis, methylation analysis, selective chemical degradation (periodate oxidation-Smith degradation and hydrazinolysis-nitrous acid deamination), enzymatic (peptide:N-glycosidase F, several beta-galactosidases, beta-hexosaminidase and alpha-galactosidase) digestions and instrumental analyses (1H-NMR and fast atom bombardment mass spectrometry) to have the novel and unique carbohydrate sequences, Gal alpha 1-->3(Gal beta 1-->4)Gal beta 1-->4GlcNAc beta 1--> and Gal alpha 1-->3(+/- GalNAc beta 1-->4GlcNAc beta 1-->3Gal beta 1-->4)Gal beta 1-->4GlcNAc beta 1-->. This study represents the first detailed investigation of the nature of bulky complex asparagine-linked penta-antennary glycans with a bisecting GlcNAc residue in glycoproteins. Expression of such bulky multiantennary glycan units on proteins may be essential during early embryogenesis.

Acetylglucosamine↗
PubMed1996

Mimicking the high levels of activity of a large in-vitro fertilization unit leads to early success at the commencement of an in-vitro fertilization and embryo transfer programme.

Infertile couples seeking treatment will endeavour to obtain the highest standards of care at a conveniently located centre. The success rated of a centre would also be an important concern. The busier in-vitro fertilization (IVF) units (> 400 cycles per year) have substantially higher pregnancy rates than those carrying out < 300 cycles per year. We report our experiences at the IVF centre, Maternity Hospital, Kuwait of mimicking the high level of activity of a large IVF unit. Infertile couples were treated in groups of 30-35 every 4-6 weeks. Patients in each group had menstrual cycles synchronized to their luteal phase using progestogens. Thereafter they were down-regulated with gonadotrophin-releasing hormone agonist (long protocol). Oocyte retrieval and embryology activities were concentrated and completed in 7-10 days in each group. This episodic high level of activity was aimed at mimicking that of a busy large IVF centre, although continuous in the latter. The overall results in terms of fertilization cleavage, implantation and clinical pregnancy rates were similar to those obtained in large IVF units. The clinical pregnancy rate per cycle was 25.8-39.1% in IVF and 46.1% in intracytoplasmic sperm injection.

Adult↗
PubMed1988

Studies on psychosomatic implications of infertility--effects of emotional stress on fertilization and implantation in in-vitro fertilization.

A case report shows the long and stony path of a couple who had to go through eight attempts at IVF in 4 years until the desired child was achieved. The psychosomatic implications of the various 'failures' of treatment become clearer when compared with the results of a psycho-endocrinological questionnaire completed by 551 patients and 115 controls since 1982. This questionnaire contains among others the Giessen personality test for the image of self, mother and father, a somatic complaints score to evaluate the severity of autonomic disorders, items for the presence and severity of possible gynaeco-endocrinological symptoms and items to evaluate the reference persons and their changes during childhood. The following results could be obtained. In a comparison of the means of 58 different somatic complaints, patients rated significantly higher than controls (P = 0.002). Women with menstrual disturbances, menstrual pains, acne or unclear skin or dry skin also suffered more from other complaints. These symptoms were also correlated with a higher score for depressive mood. Those women who were not brought up by both parents during their childhood also scored significantly higher (P less than 0.0001) in the somatic complaints score. In the Giessen personality test patients appeared significantly more compulsive (P less than 0.002) and depressive (P = 0.003) than the controls and identified themselves more with their mothers. From 60 patients in our IVF programme, 19 had no fertilization of oocytes and scored significantly higher (P = 0.008) on the social potency scale than did the patients with fertilized oocytes. Women with unsuccessful treatments in terms of pregnancy scored significantly higher in the social resonance and social potency tests. We interpret this exaggerated positive attitude as an attempt to overcome inner fears, doubts and ambivalences. These results support the importance of psychosomatic aspects in infertility and reassure us of the necessity of including counselling in any infertility treatment, including IVF.

Adult↗
PubMed1989

Immunoreactive renin variations during fertile and infertile hyperstimulated cycles with in-vitro fertilization and embryo transfer or gamete intra-fallopian transfer.

The concentrations of prorenin (PR) and active renin (AR) were determined sequentially during ovarian hyperstimulated cycles for in-vitro fertilization-embryo transfer or gamete intra-Fallopian transfer. AR and PR increased during the early luteal phase and PR was highly correlated with the progesterone concentration. In fertile cycles, AR increased more rapidly after HCG injection and AR and PR levels were significantly higher during the late luteal phase. These results confirm the role of the renin angiotensin system in ovarian physiology.

Adult↗
PubMed1993

Partial zona dissection enhances the fertilization rate in cases of sperm defect only when total failure has occurred in previous in-vitro fertilization attempts.

The value of partial zona dissection (PZD) was assessed in a randomized protocol of in-vitro fertilization (IVF) including 30 couples (44 cycles) with at least two sperm defects. Couples were divided into three groups according to previous IVF status, i.e. group I: total failure of previous IVF; group II: marginal success of previous IVF, and group III: no previous IVF. Both PZD and untreated oocytes were inseminated using the swim-across method. Although PZD did not enhance the overall proportion of fertilized eggs resulting in diploid cleaving embryos per retrieved oocyte (14.7 versus 17.1%), this rate was higher with PZD than with IVF when previous IVF attempts had failed (group I: 16.1 versus 4.5%, P < 0.01). The rate was similar for PZD and IVF in group II, and lower for PZD than IVF in group III (15.1 versus 30.8%, P < 0.01). The higher polyspermy rate associated with PZD in groups in which classical IVF worked, could partially explain these results. The role of a possible deleterious effect of PZD on the oocytes is discussed. Out of the 44 cycles, 20 reached the embryo transfer stage and four pregnancies were obtained. In our opinion PZD should be carried out in cases of sperm defects only when previous IVF attempts have failed.

Embryo Transfer↗
PubMed1993

Subzonal sperm insertion with aged human oocytes from failed in-vitro fertilization attempts: fertilization results and some applications.

This study was undertaken to assess the potential of aged human oocytes from failed in-vitro fertilization attempts as a model for the study of fertilization events after subzonal sperm insertion (SUZI). Criteria of aged oocyte suitability for this purpose were (i) the absence of nuclei, (ii) the presence of a polar body, (iii) the absence of cell division or fragmentation, (iv) marked ooplasmic contraction in hyperosmotic medium, and (v) rapid ooplasmic relaxation after returning into normo-osmotic medium following the micromanipulation. Micro-injection techniques were essentially the same as for SUZI with fresh oocytes. Oocytes that fused with the micro-injected spermatozoa developed pronuclei of typical internal structure. However, the number of pronuclei was often higher than that theoretically expected if each sperm nucleus incorporated into the oocyte gave rise to a single pronucleus. Thus, the use of aged oocytes implies the need for a specific method to assess the frequency of fusion in sperm samples examined. The results suggest that the method described here can be applied in a preliminary diagnostic test before a therapeutic SUZI attempt and in studies aimed at the optimization of sperm treatment protocols to increase the fusion capacity of subzonally inserted spermatozoa.

Acrosome↗
PubMed2003

Production of piglets derived from in vitro-produced blastocysts fertilized and cultured in chemically defined media: effects of theophylline, adenosine, and cysteine during in vitro fertilization.

To further develop defined conditions for in vitro fertilization (IVF) and in vitro culture (IVC) of in vitro-matured porcine oocytes, we evaluated the effects of theophylline, adenosine, and cysteine in a chemically defined medium during IVF. Viability to full term of in vitro-produced blastocysts after IVF and IVC in chemically defined medium was also investigated by embryo transfer to recipients. A chemically defined medium, porcine gamate medium (PGM), was modified from porcine zygote medium (PZM-4), which was previously established. PGM was used as a basal medium for IVF and PZM-4 was for the culture of presumptive zygotes. Addition of 2.5 mM theophylline to PGM significantly increased the percentage of male pronuclear formation compared with controls (no addition). Addition of 1 microM adenosine to PGM supplemented either with or without 2.5 mM theophylline significantly reduced the number of penetrated spermatozoa compared with controls (no addition of adenosine). Supplementation with 0.2 microM cysteine in PGM containing both 2.5 mM theophylline and 1 microM adenosine further increased the percentage of development to the blastocyst stage, compared with no supplementation of cysteine, but there was no difference in fertilization parameters, such as monospermy and pronuclear formation, regardless of presence or absence of theophylline and adenosine. When Day 5 blastocysts were transferred into four recipients (20-25 blastocysts per recipient), all recipients became pregnant and farrowed a total of 21 live piglets. The present results clearly demonstrate that porcine blastocysts can be produced by IVF and IVC in chemically defined media and that they can develop to full term after embryo transfer.

Adenosine↗
PubMed1984

Effect of experimentally induced renal failure upon the fertility in rats. Fertility in uremic rats.

Two groups of rats, one with surgically induced chronic renal failure and a sham-operated group were used. 10 weeks after surgery the animals were individually mated for 15 days according to 4 different schemes: uremic couple, control couple, a couple with a uremic male, and a couple with a uremic female. A second mating was allowed 18 weeks after surgery. There the uremic rats and nonfertile controls were matched against fertile controls. The experimental group showed a higher percentage of nonfertile rats, their offspring had fewer newborns and the uremic mothers had litters that weighed less at birth. In uremic males low plasma testosterone levels were detected. The analysis of plasma luteinizing hormone in the female and the delayed fertilization of uremic mothers suggest the presence of irregularities in the estrous cycle of uremic females.

Animals↗
PubMed1983

In vitro fertilization and embryo transfer: fertile areas for litigation.

Is the family primarily a biologic unit composed of a fertile male, a fertile female and children who are genetically theirs, or is the family an essentially consensual unit wherein a man and a woman who are married to each other agree to have and raise children, to regard themselves and the children as a family, and to give each other the comforts of material and emotional support, regardless of any genetic nexus?

Ethics, Medical↗
PubMed2001

Chromosomal analysis of "failed-fertilized" human oocytes resulting from in-vitro fertilization and intracytoplasmic sperm injection.

To evaluate the incidence of chromosomal abnormalities in "failed-fertilized" oocytes derived from in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) procedure, a cytogenetic analysis was performed on 164 IVF and 64 ICSI oocytes. One hundred and eleven (67.7%) of the IVF and 56 (87.5%) of the ICSI oocytes were successfully karyotyped. Of 111 IVF oocytes, 73 (65.8%) exhibited normal haploid and 38 (34.2%) were abnormal. The abnormalities included 25 aneuploid (22.5%) (7 hyperhaploid and 18 hypohaploid), 11 diploid (9.9%) and 2 structural anomalies (1.8%). Of 56 ICSI oocytes, 52 (92.8%) were normal haploid and only 4 (7.2%) were aneuploid, with 2 hyperhaploid and 2 hypohaploid. The sperm nuclei were observed in 43 IVF oocytes (38.7%), composed of 38 (34.2%) premature chromosome condensation (PCC) and 5 (4.5%) decondensed sperm heads. Evidence of successful sperm delivery was found in all 56 ICSI oocytes; 25.0 per cent (14/56) showed PCC, 17.9 per cent (10/56) showed decondensed sperm heads, and 57.1 per cent (32/56) showed intact sperm heads. This study suggested that about one-third of unfertilized oocytes exhibited chromosomal abnormalities. The difference of aneuploidy between IVF and ICSI oocytes needs further studies analysing a larger number of oocytes.

Adult↗
PubMed1992

[What results can be expected from in vitro fertilization and other technics of assisted fertilization?].

Results of assisted reproductive techniques (IV, GIFT and ZIFT) have improved progressively during the last years but they are still not satisfactory specially when they are analyzed in terms of cost efficacy. During 1990, in the world literature, the following successful results were reported: pregnancy rates of 20% and delivery rates of 15% for IVF-ET, 29% and 22% for GIFT and 21% and 16% for ZIFT. In spite of the small number of cases reported in Chile, results are similar to those of USA and Europe. Results of assisted fertilization techniques have improved with the use of Gn-RH agonists and are worse in patients older than 35 years of age than in younger ones. Pregnancies rates are higher when the number of transferred ovocytes increases although transfer of more than 3 or 4 ovocytes or embryos is contraindicated because of the risks of multiple pregnancies. When more than 3 or 4 embryos are obtained those not transferred are usually cryopreserved. IVF-ET is the first choice of treatment in cases of infertility due to severe tubal damage and/or extensive and dense pelvic adhesions. In cases of infertility with normal tubes, intrauterine inseminations in 3 to 4 controlled hyperstimulated cycles are recommended before indicating an assisted fertilization procedure. The most risky complication of assisted reproduction, besides multiple pregnancy, is severe ovarian hyperstimulation syndrome (SOHS). This syndrome is prevented by not injecting HCG or by not transferring ovocytes or embryos in cases of high estradiol levels after HMG and/or FSH administration.

Adult↗
PubMed1988

Fertilization in vitro of bovine oocytes: analysis of some factors affecting the fertilization rates.

Ovarian follicles were aspirated from superovulated heifers at different periods from the beginning of standing heat. The follicular fluid of these follicles was analyzed for progesterone, estradiol, testosterone and androstenedione concentrations and compared to the maturation stages of the oocytes and their ability to undergo fertilization in vitro. The results obtained suggest that high concentrations of progesterone, testosterone and androstenedione compared to a lower concentration of estradiol correspond to a high proportion of M-II oocytes and a high in vitro fertilization rate.

Animals↗
PubMed1994

Structural studies of a novel type of pentaantennary large glycan unit in the fertilization-associated carbohydrate-rich glycopeptide isolated from the fertilized eggs of Oryzias latipes.

In a previous report (Kitajima, K., Inoue, S., and Inoue, Y. (1989) Dev. Biol. 132, 544-553), we found the presence of a heavily glycosylated polyprotein, "H-hyosophorin," isolated from the unfertilized eggs of Oryzias latipes. We now report our detailed analysis of the structure of the N-glycan chain in L-hyosophorin, the smallest repeating unit of H-hyosophorin, which was isolated from the fertilized eggs of O. latipes and formed from H-hyosophorin upon fertilization. The N-glycan structures were defined by a combination of compositional analysis, methylation analysis, selective chemical degradation (i.e. mild methanolysis, periodate-Smith degradation, and hydrazinolysis-nitrous acid deamination), enzymatic (endo-beta-galactosidase, peptide:N-glycanase, and Newcastle disease virus sialidase) digestion, and instrumental analyses (one- and two-dimensional proton nuclear magnetic resonance spectroscopy and fast atom bombardment mass spectrometry) which revealed novel and unique features: (a) the presence of highly branched poly-N-acetylactosamino pentaantennary structures; (b) the presence of a beta-galactosylated Lewis X antigenic epitope, Gal beta 1-->4 Gal beta 1-->4 (Fuc alpha 1-->3) GlcNAc beta 1-->; (c) the presence of a beta-galactosylated sialyl Lewis X structure, Gal beta 1-->4 (Neu5Ac alpha 2-->3) Gal beta 1-->4(Fuc alpha 1-->3) GlcNAc beta 1-->; (d) the presence of Gal beta 1-->4 Gal beta 1--> and Gal beta 1--> 4Gal beta 1-->4Gal beta 1--> as the major and minor groupings, respectively; and (e) the presence of the branched Gal residues, -->4GlcNAc beta 1-->3(Gal beta 1-->4) Gal beta 1-->. This study represents the first detailed investigation regarding the nature of highly branched complex asparagine-linked pentaantennary glycans in glycoproteins. The unique expression of such bulky multiantennary glycan units on proteins could be essential during early embryogenesis.

Animals↗