Kentaro Higuchi

Development of Spermatogonial Cell Transplantation in Nibe Croaker, Nibea mitsukurii (Perciformes, Sciaenidae)

In a recently established system for intraperitoneal spermatogonial cell transplantation in salmonids, donor type A spermatogonia (type A SG) were microinjected into the peritoneal cavity of newly hatched larvae. Compared with salmonids, the larvae of marine teleosts are small and vulnerable to physiological and physical stresses, making it difficult to use them for cell manipulation. Herein, we developed type A SG cell transplantation in Nibe croaker (Nibea mitsukurii) by optimizing 1) the developmental stage of the donor testes used to prepare type A SG-enriched cell suspensions and 2) the timing and location of intraperitoneal cell transplantations to recipient larvae. Donor cells labeled with PKH26 fluorescent dye were transplanted into the peritoneal cavity of 3-, 4-, 5-, and 6-mm larvae using glass micropipettes. Consequently, 20.6% of the 4-mm larvae recipients survived for 3 wk, and 36.3% of the survivors had donor-derived cells in their gonads. The incorporated donor cells were identified as germ cells by germ cell-specific nuclear morphology and expression of a germ cell marker. In contrast, no donor type A SG were incorporated into the gonads of 6-mm recipient larvae. These data indicate that there is a distinct narrow window in the developmental stages of recipient larvae when exogenous type A SG can be incorporated into the gonads. The establishment of this system in pelagic egg-spawning marine teleosts would allow the creation of a new broodstock system in which a target species with a large body size and long generation time could be produced from related species with a small body size and short generation time.

Chub Mackerel Gonads Support Colonization, Survival, and Proliferation of Intraperitoneally Transplanted Xenogenic Germ Cells

The production of xenogenic gametes from large-bodied, commercially important marine fish species in closely related smaller host fish species with short generation times may enable rapid and simple seed production of the target species. As a first step toward this goal, we assessed the suitability of chub mackerel, Scomber japonicus, as a small-bodied recipient species for xenogenic spermatogonial transplantation. Histological observation of the early gonadal development of chub mackerel larvae and transplantation of fluorescent-labeled spermatogonia from Nibe croaker, Nibea mitsukurii, revealed that 5.3-mm chub mackerel larvae were suitable recipients for successful transplantation. Intraperitoneally transplanted xenogenic spermatogonia efficiently colonized the gonads of these recipient larvae, and donor-derived Nibe croaker germ cells proliferated rapidly soon after colonization. Moreover, gonadal soma-derived growth factor (gsdf) mRNA, a gonadal somatic cell marker, was expressed in recipient-derived cells surrounding the incorporated donor-derived germ cells, suggesting that donor-derived germ cells had settled at an appropriate location in the recipient gonad. Our data show that xenogenic spermatogonial transplantation was successful in chub mackerel and that the somatic microenvironment of the chub mackerel gonad can support the colonization, survival, and proliferation of intraperitoneally transplanted xenogenic germ cells derived from a donor species of a different taxonomic family.

Differential growth rates related to initiation of piscivory by hatchery-reared larval Pacific bluefin tuna Thunnus orientalis

Fast growth plays an important role in survival processes during the early life stages of both field-captured and hatchery-reared Pacific bluefin tuna Thunnus orientalis. Marked growth variations in hatchery-reared tuna larvae are frequently observed even for the same age and within the same rearing tank after the onset of the piscivory. We hypothesized that these small growth variations in the growth of tuna larvae at the onset of piscivory lead subsequently to large growth variations and tested the hypothesis using three size groups (large, intermediate and small) of hatchery-reared fish by nitrogen stable isotope and otolith analyses. Stable isotope analysis revealed that the large group rapidly utilized prey fish larvae, but the smaller groups depended more on rotifers as the main prey item relative to the large group. The otolith radius from the core to the increment corresponding to the first feeding on yolk-sac larvae was compared among the three size groups. The results revealed that the large group had larger otolith radii than the small and intermediate groups. Our findings suggest that small growth variations apparent during the early larval stage of tuna could induce further large growth variations in the late-larval and juvenile stages through differences in the initial ability to utilize piscivory.