Egg-binding Xenopus laevis sperm protein immunologically homologous with boar spermadhesins

0463078 - BTÚ 2017 NL eng C - Conference Paper (international conference)
Maňásková-Postlerová, Pavla - Jonáková, Věra
Egg-binding Xenopus laevis sperm protein immunologically homologous with boar spermadhesins.
Abstract book of 18th International Congress on animal reproduction. Amsterdam: Elsevier, 2016, s. 343-343.
[The 18th International Congress on Animal reproduction. Tours (FR), 26.06.2016-30.06.2016]
R&D Projects: GA ČR GA14-05547S; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:86652036
Keywords : sperm-egg interaction * Xenopus laevis * spermadhesins * enzymatic activity * glycoproteins
Subject RIV: EB - Genetics ; Molecular Biology

Sperm-egg recognition and interaction is a crucial step of fertilization in all animals. Mammalian oocyte is surrounded by the glycoprotein network called zona pellucida coded by 3-4 genes. In Xenopus laevis, the oocyte is surrounded by a vitelline envelope, which is composed of a similar glycoprotein as in mammals. In the present study, we extracted sperm proteins of X. laevis from the surface using a kit for isolation of cell surface proteins, and isolated sperm proteins by acidic extraction. We looked for the similarity of Xenopus proteins to boar sperm proteins, which are able to interact with the egg envelope. Xenopus sperm proteins were characterized by SDS-electrophoresis, and substrate-zymographic methods were used for detection of the enzymatic activity (proteolytic and hyaluronidase) in both sperm extracts. Antigenic determinants of Xenopus sperm proteins similar to mammalian sperm proteins were studied using polyclonal antibodies prepared against boar seminal plasma and sperm proteins. Polyclonal antibodies raised against boar AQN and AWN spermadhesins showed to react with a protein band of 14 kDa of the X. laevis sperm extract on the blot. This protein also interacted with biotin-labeled glycoproteins of the Xenopus vitelline envelope. Indirect immunofluorescence revealed similar antigen determinant to spermadhesins on the Xenopus sperm surface in the acrosomal cap area. This suggests that similarly as in mammals, there is a protein in X. laevis spermatozoa that is homologous to AQN and AWN spermadhesins of the pig and that also displays the binding activity for porcine zona pellucida.
Permanent Link: http://hdl.handle.net/11104/0270348