Fluorescent analysis of boar sperm capacitation process in vitro

0555425 - BTÚ 2022 RIV GB eng J - Journal Article
Děd, Lukáš - Dostálová, Pavla - Žatecká, Eva - Dorosh, Andriy - Komrsková, Kateřina - Pěknicová, Jana
Fluorescent analysis of boar sperm capacitation process in vitro.
Reproductive Biology and Endocrinology. Roč. 17, č. 1 (2019), č. článku 109. E-ISSN 1477-7827
R&D Projects: GA ČR(CZ) GA18-11275S; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:86652036
Keywords : Acrosome reaction * Fluorescent microscopy * Flow cytometry
OECD category: Reproductive biology (medical aspects to be 3)
Impact factor: 3.235, year: 2019
Method of publishing: Open access
https://rbej.biomedcentral.com/articles/10.1186/s12958-019-0554-z

Background Capacitation involves physiological changes that spermatozoa must undergo in the female reproductive tract or in vitro to obtain the ability to bind, penetrate and fertilize the egg. Up to date, several methods have been developed to characterize this complex biological process. The goal of the presented study is to mutually compare several fluorescent techniques, check their ability to detect changes in molecular processes during the capacitation progress and determine their ability to predict the percentage of acrosome reacted (AR) sperm after the exposure to solubilized zona pellucida (ZP). The capacitation process was analyzed using four fluorescent techniques: 1. chlortetracycline (CTC) staining, 2. anti-acrosin antibody (ACR.2) assay, 3. anti-phosphotyrosine (pY) antibody assay, 4. fluorescein isothiocyanate-conjugated phalloidin (FITC-phall) assay. All these methods were tested using fluorescent microscopy and flow cytometry. Results All selected methods are capable to detect the capacitation progress of boar sperm in vitro, but there are significant differences in their outcome when using fluorescent microscopy or flow cytometry experimental arrangements and subsequent statistical analysis (KW-ANOVA). Also, the ability to predict the absolute numbers of sperm which will undergo ZP-induced AR differ significantly (CTC and ACR.2 gave the best predictions). Conclusions Our study compared four largely used methods used to characterize capacitation process, highlighted their differences and showed that all are able to detect capacitation progress, CTC and ACR.2 are furthermore able to accurately predict the percentage of AR sperm after ZP-induced AR.
Permanent Link: http://hdl.handle.net/11104/0329944