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Protein-protein interactions in sperm and egg under the spotlight of super-resolution microscopy.
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SYSNO ASEP 0482835 Document Type K - Proceedings Paper (Czech conf.) R&D Document Type The record was not marked in the RIV Title Protein-protein interactions in sperm and egg under the spotlight of super-resolution microscopy. Author(s) Dvořáková-Hortová, Kateřina (BTO-N)
Děd, Lukáš (BTO-N) RID
Šebková, Nataša (BTO-N)
Frolíková, Michaela (BTO-N) ORCIDNumber of authors 4 Action Superresolution and Life cell imaging symposium 2017 Event date 05.04.2017 - 07.04.2017 VEvent location Vestec Country CZ - Czech Republic Language eng - English Keywords CD46 ; STED ; acrosome reaction Subject RIV BO - Biophysics OECD category Biophysics R&D Projects GA14-05547S GA ČR - Czech Science Foundation (CSF) ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Institutional support BTO-N - RVO:86652036 Annotation The acrosome reaction (AR) is a process of membrane fusion and lytic enzyme release, which enables sperm to penetrate the egg surroundings. It is widely recognized that specific sperm proteins form an active network prior to fertilization, and their dynamic relocation is crucial for the sperm-egg fusion. Protein CD46 is present on the acrosomal membrane of mouse sperm head and experiments with CD46-deficient mouse established the theory that CD46 participates in its stabilization. However, using super-resolution microscopy we demonstrated that CD46 is relocalizated during capacitation and acrosome reaction across the sperm head. This fact suggested that its role does not end by the loss of the acrosome and CD46 is probably a component of large multiprotein complex on sperm membrane that is responsible for sperm-egg interaction. The integrins are transmembrane proteins that participate in many cell-cell and cell-extracelular matrix interaction. The integrins are able to associate with other membrane receptors in multi-molecular complexes that participate in cell activation. Integrins are proteins consist of two different subunits α and β, when the β subunit is capable of binding to actin and controlling of its remodeling. The ability of β1 integrins to associate with CD46 is known from somatic cells. We showed that β1 integrins were present in acrosomal area of mouse sperm head and they were relocalized during capacitation and acrosome reaction similarly to CD46. Thanks to using STED and SIM super-resolution microscopy, we were able to distinguish mutual position of these protein in sperm head. This mutual position of CD46 and β1 integrins was investigated by colocalization analysis of super-resolution images and their association was confirmed by proximity ligation assay. Moreover, we presented that integrins α3 and α6 subunits were possibly pairing with β1, but occupying different compartments of the intact sperm head. Workplace Institute of Biotechnology Contact Monika Kopřivová, Monika.Koprivova@ibt.cas.cz, Tel.: 325 873 700 Year of Publishing 2018
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