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Preservation of connexin 43 and transzonal projections in isolated bovine pre-antral follicles before and following vitrification

  • Fertility Preservation
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A Correction to this article was published on 17 December 2020

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Abstract

Purpose

Gap junctions and transzonal projections play a crucial role in intercellular communication between different follicular components and are necessary for follicle development. We aimed to demonstrate gap junction protein connexin 43 (Cx43) and transzonal projections (TZPs) in viable, category 1, isolated bovine pre-antral follicles (PAFs) during short-term culture and after vitrification and warming.

Methods

This study involved four experimental groups: fresh control, 2-day culture, 4-day culture, and vitrified secondary PAFs. Isolated PAFs were vitrified using a simple and efficient cryopreservation method by means of mini cell strainers.

Results

Cx43 and TZPs were detected in pre-antral follicles of all stages, as well as in every experimental group. The group fresh follicles showed a higher percentage of follicles that were positive for Cx43 (91.7%) than the follicles that were vitrified (77.4%). All follicles that were cultured for 2 days were Cx43-positive (100%). Follicles cultured for 4 days (65.8%) (P = 0.002) showed the lowest percentage of follicles that were Cx43-positive. The percentages of the presence or (partial) absence of the TZP network were shown to be very heterogeneous between follicles in different treatment groups.

Conclusions

These results suggest the maintenance of communication between the oocyte and the somatic companion cells after vitrification and warming. The varying percentages of the expression of the TZP network within groups suggests that it will be of interest to investigate whether this is truly due to variability in TZP integrity and follicle quality or due to methodological limitations.

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Acknowledgments

The authors thank Silke Andries and Els Merckx for their excellent technical assistance and the local slaughterhouses for their cooperation in sample collection. The authors thank Dr. Bronwen Martin for reading and editing the article. The Leica SP8 (Hercules grant AUHA.15.12) confocal microscope was funded by the Hercules Foundation of the Flemish Government.

Funding

All (co-)authors state that the funding of this research was provided by the independent Operational Costs of the University of Antwerp.

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Authors and Affiliations

  1. Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Laboratory of Veterinary Physiology and Biochemistry, Gamete Research Centre, University of Antwerp, Universiteitsplein 1, U building, 2610, Wilrijk, Belgium

    Anniek Bus, Jo L. M. R. Leroy & Peter E. J. Bols

  2. Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, 310 Cedar Street, New Haven, CT, 06510, USA

    Katarzyna Szymanska

  3. Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Laboratory of Cell Biology and Histology, University of Antwerp, Universiteitsplein 1, T building, 2610, Wilrijk, Belgium

    Isabel Pintelon

  4. Faculty of Medicine and Health Sciences, Department of Basic and Applied Medical Sciences (BAMS), Physiology group, Ghent University, C. Heymanslaan 10, B-9000, Ghent, Belgium

    Katarzyna Szymanska & Luc Leybaert

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  1. Anniek Bus
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  2. Katarzyna Szymanska
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  3. Isabel Pintelon
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  4. Jo L. M. R. Leroy
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Correspondence to Anniek Bus.

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Bus, A., Szymanska, K., Pintelon, I. et al. Preservation of connexin 43 and transzonal projections in isolated bovine pre-antral follicles before and following vitrification. J Assist Reprod Genet 38, 479–492 (2021). https://doi.org/10.1007/s10815-020-01993-2

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  • DOI: https://doi.org/10.1007/s10815-020-01993-2

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