Early development of primordial germ cells in Pacific bluefin tuna Thunnus orientalis

Expression of the dead end gene during embryogenesis and its role in primordial germ cell development in Pacific bluefin tuna, Thunnus orientalis (Temminck et Schlegel)

“…Previously, it has been reported that the deduced amino acid sequence of PBTdnd possesses an RNA recognition motif (Yazawa et al, 2013). In fact, in PBT embryos, an artificial mRNA‐coding zebrafish nanos3 3’UTR was stabilized in PGCs, as shown in this study and in the study by Higuchi, Goto, et al (2019). Although further study is required, these results suggest that PBTdnd contributes to the migration and survival of PGCs via the stabilization of germline RNAs in PBT, as reported in zebrafish (Weidinger et al, 2003).…”

“…For PGC visualization, capped sense RNA coding zebrafish nanos3 3’UTR fused with green fluorescent protein (Köprunner et al, 2001) was co‐injected (300 ng/μl) with MO. Synthesis of capped sense RNA, microinjections, culture of embryos, and rearing of hatched larvae were conducted as described by Higuchi, Goto, et al (2019).…”

First Data on Sexual Dimorphic Growth of Cultured Pacific Bluefin Tuna, Thunnus orientalis (Temminck et Schlegel), and Its Sex Manipulation by Treatment with an Aromatase Inhibitor

“…Although the mechanisms underlying sex diferentiation have been well studied in various aquaculture target fshes [23,24], information on this phenomenon in tuna species is scarce, owing to the difculty in obtaining artifcial seedlings that can be used for experiments. To gain a deeper understanding of reproductive biology in tuna species, we have recently studied crucial reproductive aspects of PBT, as a model of tuna species, particularly during the early life stages, based on our closed-cycle aquaculture technique [30,59,60]. Among them, we succeeded in revealing the timing and morphological characteristics of gonadal sex diferentiation in PBT [30].…”

Targeted mutagenesis of the ryanodine receptor by Platinum TALENs causes slow swimming behaviour in Pacific bluefin tuna (Thunnus orientalis)

“…The microinjection of ryr1b -TALENs was performed using a needle (G-1, Narishige, Tokyo, Japan) with an added constriction (3‒5 μm inner diameter) in the upper part of the tip in order to control the pressure required for injecting the RNA solution into the embryo and to prevent backflow of cytoplasm from the embryo as described previously 39–41 . Embryos in the 1‒4 cell stage were placed on a 1% agar-coated Petri dish filled with 50% sterilised seawater, and then microinjected with TALEN mRNA.…”