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Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm

BACKGROUND: In order to improve ICSI, appropiate sperm selection and oocyte activation is necessary. The objective of the present study was to determine the efficiency of fertilization using ICSI with chemically activated ovine oocytes and sperm selected by swim up (SU) or swim up + zona pellucida (...

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Autores principales: Hernández-Pichardo, J. E., Ducolomb, Y., Romo, S., Kjelland, M. E., Fierro, R., Casillas, F., Betancourt, M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100180/
https://www.ncbi.nlm.nih.gov/pubmed/27826442
http://dx.doi.org/10.1186/s40104-016-0124-6
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author Hernández-Pichardo, J. E.
Ducolomb, Y.
Romo, S.
Kjelland, M. E.
Fierro, R.
Casillas, F.
Betancourt, M.
author_facet Hernández-Pichardo, J. E.
Ducolomb, Y.
Romo, S.
Kjelland, M. E.
Fierro, R.
Casillas, F.
Betancourt, M.
author_sort Hernández-Pichardo, J. E.
collection PubMed
description BACKGROUND: In order to improve ICSI, appropiate sperm selection and oocyte activation is necessary. The objective of the present study was to determine the efficiency of fertilization using ICSI with chemically activated ovine oocytes and sperm selected by swim up (SU) or swim up + zona pellucida (SU + ZP) binding. RESULTS: Experiment 1, 4–20 replicates with total 821 in vitro matured oocytes were chemically activated with ethanol, calcium ionophore or ionomycin, to determine oocyte activation (precense of one PN). Treatments showed similar results (54, 47, 42 %, respectively) but statistically differents (P < 0.05) than mechanical activated oocytes in sham, ICSI and sham injection (13, 25, 32 %, respectively) (10–17 replicates; n = 429). Experiment 2: Twelve ejaculates and 28 straws of semen were used (11–19 replicates). Sperm were selected by SU in BSA-TCM 199-H medium. A total of 2,294 fresh sperm and 2,760 from frozen-thawed semen were analyzed after SU or SU + ZP binding. Fresh sperm selected by SU showed acrosome reaction (AR) of 59 %, the sperm selected by SU + ZP binding increased AR to 91 %. In comparison, the AR of frozen-thawed sperm using SU or SU + ZP binding was 77 and 86 %, respectively (P < 0.05). Experiment 3: fertilization in 200 mechanical activativated oocytes (17 replicates) was 4 %, but fertilization increased in ethanol activated oocytes after ICSI (12-28 %) (5–6 replicates). When fresh sperm only selected by SU were injected to 123 oocytes, a fertilization rate (28 %) was achieved; in sperm selected by SU + ZP was 25 % (73 oocytes). In comparison, in frozen-thawed sperm selected by SU, fertilization was 13 % (70 oocytes), whereas sperm from SU + ZP binding displayed 12 % (51 oocytes) (P > 0.05). CONCLUSIONS: Chemical activation induces higher ovine oocyte activation than mechanical activation. Ethanol slightly displays higher oocyte activation than calcium ionophore and ionomicine. Sperm selection with SU + ZP increased AR/A and AR/D rates in comparison with SU in fresh and frozen-thawed sperm. According to this, in terms of fertilization rates, chemical activation after ICSI increased oocyte PN formation compared to mechanical activation. Also, fresh sperm treated with SU and SU + ZP were significantly different than frozen-thawed sperm, but between sperm treatments no significant differences were obtained.
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spelling pubmed-51001802016-11-08 Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm Hernández-Pichardo, J. E. Ducolomb, Y. Romo, S. Kjelland, M. E. Fierro, R. Casillas, F. Betancourt, M. J Anim Sci Biotechnol Research BACKGROUND: In order to improve ICSI, appropiate sperm selection and oocyte activation is necessary. The objective of the present study was to determine the efficiency of fertilization using ICSI with chemically activated ovine oocytes and sperm selected by swim up (SU) or swim up + zona pellucida (SU + ZP) binding. RESULTS: Experiment 1, 4–20 replicates with total 821 in vitro matured oocytes were chemically activated with ethanol, calcium ionophore or ionomycin, to determine oocyte activation (precense of one PN). Treatments showed similar results (54, 47, 42 %, respectively) but statistically differents (P < 0.05) than mechanical activated oocytes in sham, ICSI and sham injection (13, 25, 32 %, respectively) (10–17 replicates; n = 429). Experiment 2: Twelve ejaculates and 28 straws of semen were used (11–19 replicates). Sperm were selected by SU in BSA-TCM 199-H medium. A total of 2,294 fresh sperm and 2,760 from frozen-thawed semen were analyzed after SU or SU + ZP binding. Fresh sperm selected by SU showed acrosome reaction (AR) of 59 %, the sperm selected by SU + ZP binding increased AR to 91 %. In comparison, the AR of frozen-thawed sperm using SU or SU + ZP binding was 77 and 86 %, respectively (P < 0.05). Experiment 3: fertilization in 200 mechanical activativated oocytes (17 replicates) was 4 %, but fertilization increased in ethanol activated oocytes after ICSI (12-28 %) (5–6 replicates). When fresh sperm only selected by SU were injected to 123 oocytes, a fertilization rate (28 %) was achieved; in sperm selected by SU + ZP was 25 % (73 oocytes). In comparison, in frozen-thawed sperm selected by SU, fertilization was 13 % (70 oocytes), whereas sperm from SU + ZP binding displayed 12 % (51 oocytes) (P > 0.05). CONCLUSIONS: Chemical activation induces higher ovine oocyte activation than mechanical activation. Ethanol slightly displays higher oocyte activation than calcium ionophore and ionomicine. Sperm selection with SU + ZP increased AR/A and AR/D rates in comparison with SU in fresh and frozen-thawed sperm. According to this, in terms of fertilization rates, chemical activation after ICSI increased oocyte PN formation compared to mechanical activation. Also, fresh sperm treated with SU and SU + ZP were significantly different than frozen-thawed sperm, but between sperm treatments no significant differences were obtained. BioMed Central 2016-11-08 /pmc/articles/PMC5100180/ /pubmed/27826442 http://dx.doi.org/10.1186/s40104-016-0124-6 Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Hernández-Pichardo, J. E.
Ducolomb, Y.
Romo, S.
Kjelland, M. E.
Fierro, R.
Casillas, F.
Betancourt, M.
Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm
title Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm
title_full Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm
title_fullStr Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm
title_full_unstemmed Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm
title_short Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm
title_sort pronuclear formation by icsi using chemically activated ovine oocytes and zona pellucida bound sperm
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100180/
https://www.ncbi.nlm.nih.gov/pubmed/27826442
http://dx.doi.org/10.1186/s40104-016-0124-6
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