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Plasma membrane and acrosome loss before ICSI is required for sheep embryonic development

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

This study aims to determine if the integrity of the sperm plasma membrane and acrosome vesicle could be limiting factors in sheep intracytoplasmic sperm injection (ICSI).

Methods

Prior to in vitro fertilization (IVF) or ICSI, the oocytes were subjected to in vitro maturation (IVM) for 24 h. First, to evaluate the need of artificial activation for ovine ICSI, 226 oocytes were injected with intact spermatozoa (IS), from which 125 were activated by incubation in ionomycin and 101 were cultured without activation. Next, spermatozoa were mechanically (by piezo-electrical pulses) and/or chemically (by ionomycin/Triton X-100) treated to break membranes and acrosomes and were injected into oocytes, grouped as follows: (i) piezo-pulsed spermatozoa (PPS), (ii) PPS pre-treated with ionomycin (PPS-I), (iii) PPS pre-treated with Triton X-100 (PPS-T), and (iv) intact and untreated spermatozoa as a control (CTR-IS).

Results

No differences were observed in the zygote/cleavage/blastocyst rate between chemically activated and non-activated oocytes (50 vs. 45 %, 11.6 vs. 10.1 %; 1.8 vs. 1.1 %, respectively), after ICSI with CTR-IS. Injection of PPS compared to CTR-IS increased the proportion of zygotes and blastocysts (84.6 vs. 45 %, p < 0.01; 15.5 vs. 1.1 %, p < 0.0001, respectively). Moreover, the percentage of PPS-derived blastocysts was not significantly different from that obtained by conventional IVF (15.5 vs. 20.2 %). The ICSI blastocysts’ development was also improved with PPS pre-treated with ionomycin (15.6 %), but was completely impeded with PPS pre-treated with Triton X-100 (0 %).

Conclusion

Our findings confirm that ICSI with spermatozoa whose plasma membrane and acrosome have been mechanically damaged substantially improves embryonic development until the blastocyst stage.

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Acknowledgments

This work was supported by Program FIRB GA B81J12002520001 (GenHome) to PL, Program OVI-TE-CA 2011 (financing-Fondazione Tercas) to GP, and Program P.O. FSE Abruzzo “007-2013” to DAA. The authors are participating in the COST action FA 1201 “Epiconcept” Epigenetic and Peri-conception Environment. A warm acknowledgement goes to Dr. Nicola Ferri, Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Caporale” Campo Boario (IZSAM), Teramo, Italy, for his technical and scientific support.

Authors’ contributions

DAA, DI, and PL conceived the experiments. DAA and DI performed all experiments. DAA, DI, and PL wrote the manuscript. MC discussed the results and edited the manuscript. GP supervised the embryological work.

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Author notes

  1. Grazyna Ptak

    Present address: National Research Institute of Animal Production, 1, Krakowska Street, 32-083, Balice n/Krakow, Poland

Authors and Affiliations

  1. Faculty of Veterinary Medicine, University of Teramo, R. Balzarini Street 1, Campus Coste Sant’Agostino, 64100, Teramo, Italy

    Debora A. Anzalone, Domenico Iuso, Marta Czernik, Grazyna Ptak & Pasqualino Loi

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  1. Debora A. Anzalone
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  2. Domenico Iuso
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  3. Marta Czernik
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Corresponding author

Correspondence to Pasqualino Loi.

Additional information

Capsule Intracytoplasmic sperm injection (ICSI) with spermatozoa whose plasma membrane and acrosome have been mechanically damaged substantially improves sheep embryonic development until the blastocyst stage.

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Anzalone, D.A., Iuso, D., Czernik, M. et al. Plasma membrane and acrosome loss before ICSI is required for sheep embryonic development. J Assist Reprod Genet 33, 757–763 (2016). https://doi.org/10.1007/s10815-016-0709-1

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  • DOI: https://doi.org/10.1007/s10815-016-0709-1

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