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Fluorescent analysis of boar sperm capacitation process in vitro
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SYSNO ASEP 0555425 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Fluorescent analysis of boar sperm capacitation process in vitro Tvůrce(i) Děd, Lukáš (BTO-N) RID
Dostálová, Pavla (BTO-N)
Žatecká, Eva (BTO-N)
Dorosh, Andriy (BTO-N) RID
Komrsková, Kateřina (BTO-N) ORCID
Pěknicová, Jana (BTO-N) RIDCelkový počet autorů 6 Číslo článku 109 Zdroj.dok. Reproductive Biology and Endocrinology. - : BioMed Central
Roč. 17, č. 1 (2019)Poč.str. 11 s. Jazyk dok. eng - angličtina Země vyd. GB - Velká Británie Klíč. slova Acrosome reaction ; Fluorescent microscopy ; Flow cytometry Vědní obor RIV EB - Genetika a molekulární biologie Obor OECD Reproductive biology (medical aspects to be 3) CEP GA18-11275S GA ČR - Grantová agentura ČR ED1.1.00/02.0109 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Způsob publikování Open access Institucionální podpora BTO-N - RVO:86652036 UT WOS 000512094000001 EID SCOPUS 85077085464 DOI 10.1186/s12958-019-0554-z Anotace 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. Pracoviště Biotechnologický ústav Kontakt Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Rok sběru 2022 Elektronická adresa https://rbej.biomedcentral.com/articles/10.1186/s12958-019-0554-z
Počet záznamů: 1