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The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization

In a previous study, we reported that porcine sperm cysteine-rich secretory protein 2 (CRISP2) is localized in the post-acrosomal sheath-perinuclear theca (PT) as reduction-sensitive oligomers. In the current study, the decondensation and removal of CRISP2 was investigated during in vitro sperm capa...

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Autores principales: Zhang, Min, Bromfield, Elizabeth G, Helms, J Bernd, Gadella, Bart M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9663942/
https://www.ncbi.nlm.nih.gov/pubmed/36054334
http://dx.doi.org/10.1093/biolre/ioac169
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author Zhang, Min
Bromfield, Elizabeth G
Helms, J Bernd
Gadella, Bart M
author_facet Zhang, Min
Bromfield, Elizabeth G
Helms, J Bernd
Gadella, Bart M
author_sort Zhang, Min
collection PubMed
description In a previous study, we reported that porcine sperm cysteine-rich secretory protein 2 (CRISP2) is localized in the post-acrosomal sheath-perinuclear theca (PT) as reduction-sensitive oligomers. In the current study, the decondensation and removal of CRISP2 was investigated during in vitro sperm capacitation, after both the induction of the acrosome reaction and in vitro fertilization. Confocal immunofluorescent imaging revealed that additional CRISP2 fluorescence appeared on the apical ridge and on the equatorial segment (EqS) of the sperm head following capacitation, likely due to cholesterol removal. After an ionophore A23187-induced acrosome reaction, CRISP2 immunofluorescence disappeared from the apical ridge and the EqS area partly not only owing to the removal of the acrosomal shroud vesicles, but to its presence in a subdomain of EqS. The fate of sperm head CRISP2 was further examined post-fertilization. In vitro matured porcine oocytes were co-incubated with boar sperm cells for 6–8 h and the zygotes were processed for CRISP2 immunofluorescent staining. Notably, decondensation of CRISP2, and thus of the sperm PT, occurred while the sperm nucleus was still fully condensed. CRISP2 was no longer detectable in fertilized oocytes in which sperm nuclear decondensation and paternal pronucleus formation were apparent. This rapid dispersal of CRISP2 in the PT is likely regulated by redox reactions for which its cysteine-rich domain is sensitive. Reduction of disulfide bridges within CRISP2 oligomers may be instrumental for PT dispersal and elimination.
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spelling pubmed-96639422022-11-14 The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization Zhang, Min Bromfield, Elizabeth G Helms, J Bernd Gadella, Bart M Biol Reprod Research Article In a previous study, we reported that porcine sperm cysteine-rich secretory protein 2 (CRISP2) is localized in the post-acrosomal sheath-perinuclear theca (PT) as reduction-sensitive oligomers. In the current study, the decondensation and removal of CRISP2 was investigated during in vitro sperm capacitation, after both the induction of the acrosome reaction and in vitro fertilization. Confocal immunofluorescent imaging revealed that additional CRISP2 fluorescence appeared on the apical ridge and on the equatorial segment (EqS) of the sperm head following capacitation, likely due to cholesterol removal. After an ionophore A23187-induced acrosome reaction, CRISP2 immunofluorescence disappeared from the apical ridge and the EqS area partly not only owing to the removal of the acrosomal shroud vesicles, but to its presence in a subdomain of EqS. The fate of sperm head CRISP2 was further examined post-fertilization. In vitro matured porcine oocytes were co-incubated with boar sperm cells for 6–8 h and the zygotes were processed for CRISP2 immunofluorescent staining. Notably, decondensation of CRISP2, and thus of the sperm PT, occurred while the sperm nucleus was still fully condensed. CRISP2 was no longer detectable in fertilized oocytes in which sperm nuclear decondensation and paternal pronucleus formation were apparent. This rapid dispersal of CRISP2 in the PT is likely regulated by redox reactions for which its cysteine-rich domain is sensitive. Reduction of disulfide bridges within CRISP2 oligomers may be instrumental for PT dispersal and elimination. Oxford University Press 2022-09-02 /pmc/articles/PMC9663942/ /pubmed/36054334 http://dx.doi.org/10.1093/biolre/ioac169 Text en © The Author(s) 2022. Published by Oxford University Press behalf of Society for the Study of Reproduction. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Zhang, Min
Bromfield, Elizabeth G
Helms, J Bernd
Gadella, Bart M
The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization
title The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization
title_full The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization
title_fullStr The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization
title_full_unstemmed The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization
title_short The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization
title_sort fate of porcine sperm crisp2 from the perinuclear theca before and after in vitro fertilization
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9663942/
https://www.ncbi.nlm.nih.gov/pubmed/36054334
http://dx.doi.org/10.1093/biolre/ioac169
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