A.R. Jimenez-Macedo

Institut Marqués, Spain, Barcelona, Barcino, Catalonia, Spain

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Publications (18)44.11 Total impact

  • C. Giménez · E. Garcia-Guixé · A. Jiménez-Macedo · C. Arjona · E. Balius · M. Sandalinas

    No preview · Article · May 2013 · Reproductive biomedicine online
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    Full-text · Article · Sep 2012 · Reproductive biomedicine online
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    ABSTRACT: Developmental competence of oocytes from prepubertal females is lower than those from adult females. Oocyte development competence is positively related to follicular diameter. Most of the follicles of prepubertal goat ovaries are smaller than 3 mm. The aim of this study was to compare oocytes of two follicle sizes (< 3 mm and ≥ 3 mm) from prepubertal goats with oocytes from adult goats in relation to their in vitro production and quality of blastocysts. Oocytes from prepubertal goats were obtained from slaughterhouse ovaries and selected according to the follicle diameter whereas oocytes from adult goats were recovered in vivo by LOPU technique without prior selection of follicle size. COCs were IVM for 27 h, IVF at the conventional conditions with fresh semen and presumptive zygotes were cultured in SOF medium for 8 days. Blastocysts obtained were vitrified and after warming their blastocoele re-expansion and the ploidy by FISH technique were assessed. We found significant differences between blastocysts yield of oocytes recovered from follicles smaller than 3 mm of prepubertal goats compared to those from adult goats (5.45% vs 20. 83%, respectively) however, these differences disappear if oocytes were recovered form large follicles (18.07%). A total of 28 blastocysts were analysed and 96.43% showed mixoploidy. Age did not affect the number of embryos with abnormal ploidy or blastocyst re-expansion after warming. Furthermore, the percentage of diploid blastomeres per embryo was similar in the 3 groups studied, adult, prepubertal from follicles ≥ 3 mm and < 3 mm (68.6%, 80.8% and 73.6%, respectively). In conclusion, IVP of blastocysts coming from follicles larger than 3 mm of goats 45 days old were not different to the blastocysts produced from adult goats, both in terms of quantity and quality.
    No preview · Article · Feb 2011 · Theriogenology
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    ABSTRACT: Introduction: Whether the ART procedure has any significant effect on gender is debatable. With the introduction of a new concept for high magnification sperm morphology examination, it is now possible to select normal spermatozoa in real time for use in an intracytoplasmic morphologically selected sperm injection (IMSI) cycle. Sperm selection performed by this methodology has been showing a significant and positive association with ART outcomes. Nonetheless, there is no available literature supporting sex ratio in embryos derived from this technique. Our study’s objective was to determine whether the use of IMSI in preimplantation genetic screening (PGS) cycles is associated with gender incidence. Material and Methods: This prospective study was conducted on couples who underwent their first IVF treatment in conjunction with PGS as a result of advanced maternal age. Couples were randomly allocated to receive one of two sperm selection procedures: ICSI (n = 60) or IMSI (n = 60). Patients’ randomization was performed by laboratory personnel just before oocyte injection using SAS System for Windows (PROC PLAN, seed = 1234). To minimize the influence of male factor infertility, all cases in which sperm concentration was less than 1×106 M/mL and sperm motility less than 20% were excluded from the study. Sperm selection in IMSI group was performed using an inverted Nikon Diaphot microscope (Eclipse TE 300; Nikon®, Tokyo, Japan) equipped with high-power differential interference contrast optics (DIC/Nomarski). The total calculated magnification was 6600x. The sperm cells exhibiting normally shaped nuclei ((i) smooth, (ii) symmetric and (iii) oval configuration) and (iv) normal nuclear chromatin content (if it contained no more than one vacuole, which occupies < 4% of the nuclear area) were selected for injection. Embryos reaching at least the 6-cell stage on day 3 of development were biopsied and a two-round FISH procedure was performed, which allowed for the detection of chromosomes X, Y, 13, 15, 16, 18, 21 and 22. Embryo transfer was performed on day 5. The influence of the sperm selection method on aneuploidy rates was assessed using logistic regression analysis. The data are presented as Odds Ratios (OR) with 95% confidence interval (95% CI ) and p value. The results were considered to be significant at the 5% critical level (p < 0.05). Results: No significant differences were observed between all groups in terms of patient demographics, stimulation and cycle characteristics. Clinical pregnancy (47.1% vs 53.8%, p = 0.5321) and implantation rates (40.9% vs 45.6%, p = 0.5911) were also not significantly different. There was no abortion occurrence in IMSI cycles and only one case described in ICSI cycles. A total of 153 successfully biopsied embryos derived from 60 ICSI-PGS and 240 successfully biopsied embryos derived from 60 IMSI-PGS cycles were characterized by gender. Considering all the biopsied embryos characterized as normal for sex chromosome, our results showed that IMSI approach results in a significantly higher incidence of female embryos when compared to ICSI (64.7% vs 53.8%, respectively, p = 0.0268). Analysing only euploid embryos for the 8 analysed chromosomes we also observed a significantly higher incidence of XX embryos derived from IMSI compared to ICSI cycles (66.7% vs 52.4%, respectively, p = 0.0326). This result was confirmed by logistic regression, which demonstrated a nearly two-fold increase in euploid XX embryos derived from sperm selected by high magnification (OR: 1.82, CI: 1.05–3.15, p = 0.033). Conclusion: To our knowledge this is the first study on the sex ratio of embryos derived from IMSI cycles. “Best looking” spermatozoa analyzed by high magnification approach seem to carry a higher proportion of X chromosome suggesting a significantly predictive value on the incidence of euploid XX embryos.
    Full-text · Article · Jan 2011 · Human Reproduction
  • R Romaguera · R Morató · A.R. Jiménez-Macedo · Catalá MG · M Roura · M.T. Paramio · M.J. Palomo · T Mogas · D Izquierdo
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    ABSTRACT: Oocytes secrete soluble paracrine factors called Oocyte Secreted Factors (OSFs) which regulate the cumulus cell phenotype. Follicle populations in ovaries from prepubertal females have smaller diameters than their adult counterparts. Oocytes from small follicles are less competent than those from large follicles. The aim of this study was to investigate, in prepubertal goats, the effect of OSFs secreted by denuded oocytes (DOs) from small (< 3 mm) (>= 3 mm) follicles during IVM on embryo development and the blastocyst quality of cumulus-oocyte complexes (COCs) from small follicles and to determine if GDF9 participates in this process. Treatment groups were: (A) COCs non selected by their follicle size (control group); (B) cumulus oocytes complexes from small follicles (SFCOCs),
    No preview · Article · Oct 2010 · Theriogenology
  • R Romaguera · A Casanovas · R Morató · D Izquierdo · M Catalá · A.R. Jimenez-Macedo · T Mogas · M T Paramio
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    ABSTRACT: The aim of this study was to assess the following parameters in prepubertal goat oocytes of different follicle diameter (> or =3 mm, <3 mm, control): oocyte diameter, early (Annexin-V) and late (TUNEL) apoptosis, embryo development and chromosomal ploidy of these blastocysts using Fluorescence In Situ Hybridization (FISH). Before in vitro maturation, oocytes were measured and stained with Annexin-V or TUNEL. The rest of the oocytes were matured, fertilized, and cultured in vitro for 8 days. Oocytes from follicles of > or =3 mm showed greater mean oocyte diameter (128.27 +/- 7.20 microm vs. 125.35 +/- 7.59 microm), higher percentages of TUNEL positive (42.86 vs. 24.23%), higher cleavage (47.85 +/- 3.98 vs. 23.07 +/- 2.44 %) and blastocyst rates (19.77 +/- 3.04 vs. 4.11 +/- 1.10 %) than oocytes from follicles of <3 mm.. Blastocyst mean cell numbers did not show differences between follicular groups (123.83 +/- 49.62 vs. 104.29 +/- 36.09 for follicles of > or =3 mm and <3 mm, respectively). A total of 54 blastocysts with 7084 nuclei were hybridized with specific probes to chromosomes X and Y. Ninety-eight percent (98%) of the embryos presented at least one cell carrying an abnormal number of chromosomes, but 78% of them presented less than 25% of chromosomal abnormal cells. No differences in the percentage of blastocysts with abnormal ploidy were found in embryos produced from oocytes of different follicle diameter.
    No preview · Article · Aug 2010 · Theriogenology

  • No preview · Article · Jan 2010 · Reproduction Fertility and Development
  • B Anguita · M.T. Paramio · R Morató · R Romaguera · A.R. Jiménez-Macedo · T Mogas · D Izquierdo
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    ABSTRACT: Oocyte quality is the main factor that determines blastocyst yield; any factor that could affect it, such as apoptosis, could impair subsequent embryonic development. Our aim was to investigate the incidence of apoptosis in prepubertal goat oocytes and cumulus cells, assessed by Annexin-V staining and TUNEL assay, and their effect on embryo development. Oocyte-cumulus complexes (COCs) from slaughtered females were collected and classified depending on COC morphology as: Healthy (H) and Early Atretic (EA). Each one of these groups was classified depending on oocyte diameter: A: 110-125microm, B: 125-135microm and C: >135microm. The COCs were IVM for 27h, IVF with fresh semen and IVC for 8 days after insemination. Apoptosis analyses were performed before and after maturation. Annexin-positive oocytes decreased with diameter in the EA class (immature oocytes: A: 42.6%; B: 30.3%; C: 21%; IVM-oocytes: A: 17.5%; B: 4.8%; C: 0%), while TUNEL assay showed a decrease of apoptosis in the largest oocytes before and after IVM only in Healthy oocytes (immature oocytes: A: 51.5%; B: 43.3%; C: 12.1%; IVM-oocytes: A: 31.7%; B: 12%; C: 0%). Blastocyst rate increased with increasing oocyte diameter, and it was higher in H than in EA oocytes (Healthy; A: 0%; B: 5.3%; C: 14.4%; Early atretic: A: 0.3%; B: 4.1%; C: 5.1%). Oocyte diameter and COC morphology had no effect on the percentage of apoptosis in blastocyst cells. In conclusion, oocyte developmental competence in prepubertal goats is influenced by oocyte diameter and COC morphology.
    No preview · Article · Nov 2009 · Animal reproduction science
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    M Sánchez-Castro · A.R. Jimenez-Macedo · M Sandalinas · J Blanco
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    ABSTRACT: Sperm aneuploidy screening is now widely used as a counseling tool before performing a PGD cycle in infertile patients. The goal of this study was to determine whether sperm aneuploidy has a direct influence on embryo chromosomal abnormalities. Two groups were set up: (i) study group consisting of 13 oocyte-donation PGD cycles from males with normal karyotype and a previous altered sperm fluorescence in situ hybridization (FISH) result and (ii) control group including nine cycles of PGD for X-linked disease with no fertility problems and maternal age <36 years. Sperm samples and Day 3 embryos were evaluated using FISH for chromosomes X, Y, 13, 18 and 21. A total of 179 embryos were analyzed: 91 embryos for the control group versus 88 for the study group. The study group presented more abnormal embryos than the control group (51.14% versus 35.16%; P = 0.0353). Patients from the study group were then classified according to sperm count. Oligozoospermic patients showed a much higher proportion of abnormal embryos compared with the control group (64.87% versus 35.16%; P = 0.0030). Sperm aneuploidy and diploidy screening seems to be an effective prognostic tool that would be useful in the reproductive genetic counseling of infertile couples, especially in oligozoospermic patients.
    Full-text · Article · Feb 2009 · Human Reproduction
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    ABSTRACT: The aim of this study was to examine the relationship between the developmental competence of oocytes and their total RNA and protein contents, and the level of Cyclin B1 transcription. Ovaries from prepubertal goats were collected from a slaughterhouse. Oocytes were recovered by slicing and those with two or more layers of cumulus cells and homogenous cytoplasm were matured in vitro (20-25 oocytes per drop) for 27 h. Both before and after IVM, samples of oocytes were denuded and categorised into four group treatments by diameter (<110 microm, 110-125 microm, 125-135 microm; >135 microm), separated into sub-groups of 10 oocytes per treatment-replicate and stored in liquid nitrogen until total RNA content analysis by spectophotometry, total protein content analysis by a colorimetric assay and Cyclin B1 transcription analysis by RT-PCR. For the study of developmental competence, the rest of the matured oocytes were fertilised in vitro in groups of 20-25 for 24 h. Presumptive zygotes were denuded, sorted into the four categories of diameter noted above, and placed into culture drops in groups of 18-25 for in vitro culture. Cleavage rate was evaluated at 48 hpi and embryo development at 8 d post-insemination. There were four replicates of each treatment for each assay or evaluation point of the experiment. There were no significant differences between the size categories of oocytes at collection in total RNA content, total protein content and Cyclin B1 mRNA. There were significant differences (P<0.05) in the expression of Cyclin B1 before IVM with oocytes in the >135 mm diameter category having the highest value for this variant. There were no significant differences in these characteristics between the categories of oocyte diameter after IVM except in respect of total RNA content, which was lower for the largest size of oocytes (>135 microm; mean+/-S.D.=12.3+/-1.84 ng/oocyte) than the other three size groups (19.2+/-1.38-22.1+/-4.44 ng/oocyte; P<0.05). Significant differences (P<0.05) in cleavage rate were observed between the different oocyte size categories (<110 microm, 3.0%; 110-125 microm, 32%; 125-135 microm, 50%; >135 microm, 73%). Only oocytes >125 microm diameter developed to the blastocyst stage (125-135 microm, 7%; >135 microm, 10%). This study showed that the RNA content and the Cyclin B1 RNA expression of prepubertal goat oocytes, and their development to embryos varied between the different size categories of the oocytes.
    No preview · Article · Feb 2008 · Animal Reproduction Science
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    ABSTRACT: The purpose of this study was to determine the efficacy of pre-treating mature bovine oocytes with Taxol before vitrification by the open pulled Straw method (OPS). We evaluated the effects of pre-treating the oocytes with 1 microM Taxol on chromosome organization, spindle morphology, cortical granule distribution and the ability of fertilized oocytes to develop to the blastocyst stage. After calf or cow oocyte vitrification without Taxol, significantly higher proportions of spindle abnormalities in the form of abnormal spindle structures or dispersed or decondensed chromosomes were observed compared to fresh control oocytes. In contrast, when we compared calf oocytes pre-treated with Taxol before vitrification with control calf oocytes, similar percentages of oocytes showing a normal spindle morphology were observed. The percentages of oocytes with a peripheral cortical granule (CG) distribution increased when the oocytes were pretreated with Taxol and vitrified, while oocytes vitrified without Taxol pre-treatment gave rise to higher cortical distribution percentages. Cleavage and blastocyst rates were significantly lower for vitrified versus untreated oocytes, both in cow and calf oocytes. Significantly higher cleavage rates were obtained when calf and cow oocytes were vitrified with Taxol. Pre-treatment with Taxol before cow oocyte vitrification yielded significantly higher blastocyst rates. Calf oocytes, however, were unable to develop to the blastocyst stage, irrespective of previous Taxol treatment. These results indicate that the pre-treatment of oocytes with Taxol before vitrification helps to reduce the damage induced by the cryopreservation process, and potentially improves the subsequent development of vitrified bovine oocytes. Summary sentence: Pre-treatment of oocytes with Taxol before vitrification helps to reduce the damage induced by vitrification and potentially improves the development of vitrified bovine oocytes.
    No preview · Article · Jan 2008 · Molecular Reproduction and Development
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    ABSTRACT: The aim of this study was to analyze the relationship between oocyte diameter, meiotic and embryo developmental competence and the expression of the catalytic subunit of MPF, the p34(cdc2), at mRNA, RNA and protein level, as well as its kinase activity, in prepubertal (1-2 months old) goat oocytes. MPF is the main meiotic regulator and a possible regulator of cytoplasmic maturation; therefore, it could be a key factor in understanding the differences between competent and incompetent oocytes. Oocytes were classified according to oocyte diameter in four categories: <110, 110-125, 125-135 and >135 microm and matured, fertilized and cultured in vitro. The p34(cdc2) was analyzed in oocytes at the time of collection (0 h) and after 27 h of IVM (27 h) in each of the oocyte diameter categories. The oocyte diameter was positively related to the percentage of oocytes at MII after IVM (0, 20.7, 58 and 78%, respectively) and the percentage of blastocysts obtained at 8 days postinsemination (0, 0, 1.95 and 12.5%, respectively). The expression of RNA and mRNA p34(cdc2) did not vary between oocyte diameters at 0 and 27h. Protein expression of p34(cdc2) increased in each oocyte category after 27 h of maturation. MPF activity among diameter groups did not vary at 0h but after IVM there was a clear and statistically significant increase of MPF activity in the biggest oocytes.
    No preview · Article · Feb 2007 · Theriogenology
  • B. Anguita · M. T. Paramio · A. R. Jimenez-Macedo · R. Romaguera · R. Morato · T. Mogas · D. Izquierdo

    No preview · Article · Jan 2007 · Reproduction Fertility and Development
  • A. R. Jimenez-Macedo · B. Anguita · D. Izquierdo · R. Romaguera · R. Morato · T. Mogas · M. T. Paramio

    No preview · Article · Jan 2007 · Reproduction Fertility and Development
  • Ana Raquel Jimenez-Macedo · Dolors Izquierdo · Aixa Urdaneta · Begoña Anguita · Maria-Teresa Paramio
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    ABSTRACT: The low number of embryos obtained from IVM-IVF-IVC of prepubertal goat oocytes could be due to an incomplete cytoplasmic maturation. Roscovitine (ROS) inhibits MPF and MAP kinase activity and maintains the oocyte at Germinal Vesicle (GV) stage. The aim of this study was to determine if meiotic activity is arrested in prepubertal goat oocytes cultured with 0, 12.5, 25, 50 and 100 microM of ROS for 24 h. A group of oocytes from adult goats was cultured with 25 microM of ROS to compare the effect of ROS on prepubertal and adult goat oocytes. A sample of oocytes was stained to evaluate the nuclear stage at oocyte collection time and after ROS incubation. IVM-oocytes not exposed to ROS formed the control group. Prepubertal goat IVM-oocytes were inseminated and cultured for 8 days. The percentage of oocytes at GV stage, after exposition to ROS was significantly higher in adult goat oocytes (64.5%) than in prepubertal goat oocytes. No differences were found among 25, 50 and 100 microM ROS concentrations (29, 23 and 26%, oocytes at GV stage, respectively). After 8 days of culture, no differences in total embryos were observed between control oocytes and oocytes treated with 12.5 and 25 microM (45.2, 36.1 and 39.4%, respectively), however the percentage of blastocysts was higher in the control group. Western blot for the MAPK and p34(cdc2) showed that both enzymes were active in prepubertal goat oocytes after 24h of ROS exposition. In conclusion, a low percentage of prepubertal goat oocytes reached GV stage after ROS incubation; possibly because most of them had reinitiated the meiosis inside the follicle. ROS did not affect fertilization or total embryos but ROS showed a negative effect on blastocyst development.
    No preview · Article · Jul 2006 · Theriogenology
  • B. Anguita · A. R. Jimenez-Macedo · D. Izquierdo · M. T. Paramio

    No preview · Article · Jan 2006 · Reproduction Fertility and Development
  • B. Anguita · A.R. Jimenez-Macedo · D. Izquierdo · M.T. Paramio

    No preview · Article · Dec 2004 · Reproduction Fertility and Development
  • A.R. Jimenez-Macedo · B. Anguita · D. Izquierdo · M.T. Paramio

    No preview · Article · Dec 2004 · Reproduction Fertility and Development

Publication Stats

160 Citations
44.11 Total Impact Points

Institutions

  • 2009-2013
    • Institut Marqués, Spain, Barcelona
      Barcino, Catalonia, Spain
  • 2006-2011
    • Autonomous University of Barcelona
      • • Faculty of Veterinary
      • • Department of Animal and Food Science
      Cerdanyola del Vallès, Catalonia, Spain