Marta Blanes-García, Zoran Marinović, Juan Germán Herranz-Jusdado, Xuan Xie, Leonor Ferrão, Victor Gallego, Luz Pérez, Abdul Rasheed Baloch, Ákos Horváth, Martin Pšenička, Juan F. Asturiano, Marina Morini
Abstract
Identification of specific molecular markers for spermatogonial stem cells in teleost, is crucial for enhancing the efficacy of reproductive biotechnologies in aquaculture, such as transplantation and surrogate production in fishes. Since it is not yet possible to distinguish spermatogonial stem cells of European eel (Anguilla anguilla) using specific molecular markers, we isolated spermatogonial cells from immature European eels to find these potential markers. We attempted this by studying three candidate genes: vasa, nanos2 and dnd1. Two vasa (vasa1 and vasa2) genes, nanos2 and dnd1, were identified, characterized, and studied in muscle, testis and isolated spermatogonia. Our results showed that vasa1 and vasa2 had the highest levels of expression when measured by qPCR. In situ hybridization and immunochemistry assays showed that the four genes were localized explicitly in type A spermatogonia. However, vasa1 and vasa2 exhibited stronger signals in the immature testicular tissue than the other two potential markers. According to this, vasa1 and vasa2 were found to be the most effective markers for spermatogonial cells in European eel.
