Ubiquitin-proteasome system participates in the de-aggregation of spermadhesins and DQH protein during boar sperm capacitation

0510859 - BTÚ 2020 RIV GB eng J - Článek v odborném periodiku
Zigo, M. - Jonáková, Věra - Maňásková-Postlerová, Pavla - Kerns, K. - Sutovsky, P.
Ubiquitin-proteasome system participates in the de-aggregation of spermadhesins and DQH protein during boar sperm capacitation.
Reproduction. Roč. 157, č. 3 (2019), s. 283-295. ISSN 1470-1626
Grant CEP: GA ČR(CZ) GA18-11275S; GA MŠMT(CZ) ED1.1.00/02.0109
Institucionální podpora: RVO:86652036
Klíčová slova: SEMINAL PLASMA-PROTEINS * SURFACE-PROTEINS * ZONA-PELLUCIDA
Obor OECD: Reproductive biology (medical aspects to be 3)
Impakt faktor: 3.206, rok: 2019
Způsob publikování: Open access
https://rep.bioscientifica.com/view/journals/rep/157/3/REP-18-0413.xml

We studied the participation of the ubiquitin-proteasome system (UPS) in spermadhesin release during in vitro capacitation (IVC) of domestic boar spermatozoa. At ejaculation, boar spermatozoa acquire low molecular weight (8-16 kDa) seminal plasma proteins, predominantly spermadhesins, aggregated on the sperm surface. Due to their arrangement, such aggregates are relatively inaccessible to antibody labeling. As a result of de-aggregation and release of the outer layers of spermadhesins from the sperm surface during IVC, antibody labeling becomes feasible in the capacitated spermatozoa. In vivo, the capacitation-induced shedding of spermadhesins from the sperm surface is associated with the release of spermatozoa from the oviductal sperm reservoir. We took advantage of this property to perform image-based flow cytometry to study de-aggregation and shedding of boar spermadhesins (AQN, AWN, PSP protein families) and boar DQH (BSP1) sperm surface protein which induces higher fluorescent intensity in capacitated vs ejaculated spermatozoa. Addition of a proteasomal inhibitor (100 pM MG132) during IVC significantly reduced fluorescence intensity of all studied proteins (P< 0.05) compared to vehicle control IVC. Western blot detection of spermadhesins did not support their retention during IVC with proteasomal inhibition (P> 0.99) but showed the accumulation of DQH (P=0.03) during IVC, compared to vehicle control IVC. Our results thus demonstrate that UPS participates in the de-aggregation of spermadhesins and DQH protein from the sperm surface during capacitation, with a possible involvement in sperm detachment from the oviductal sperm reservoir and/or sperm-zona pellucida interactions. The activity of sperm UPS modulates de-aggregation of boar spermadhesins and DQH sperm surface protein/binder of sperm1 (BSP1) during the sperm capacitation.
Trvalý link: http://hdl.handle.net/11104/0301244