Laboratory of Fish Biotechnology, National Center for Research and Conservation of Continental Fish, Chico Mendes Institute of Biodiversity Conservation, Pirassununga, São Paulo, Brazil.
Institute of Bioscience, São Paulo State University, Botucatu, Brazil.
Int J Dev Biol. 2023;67(2):39-48. doi: 10.1387/ijdb.230009gc.
Biotechniques, including surrogate propagation derived from primordial germ cell (PGC) transplantation, are valuable tools for the reconstitution of endangered fish species. Although promising, there are no previous studies reporting such approaches using neotropical fish species. The aim of this study was to establish germline chimeras in neotropical fish by using the yellowtail tetra as a model species of the order Characiformes. Germline chimeras were obtained after transplantation of PGCs cultivated under different conditions: saline medium and supplemented with DMEM, amino acids, vitamins, glutamine, pyruvate, and fetal bovine serum, and subsequently transplanted into triploids and triploid hybrids from the cross between (♀) and (♂). The results indicate ectopic migration in host embryos after transplantation of PGCs cultivated in saline medium. However, PGCs cultivated in supplemented medium migrated to the region of the gonadal ridge in 4.5% of triploid and 19.3% in triploid hybrid. In addition, the higher expression of and genes was found in PGCs cultivated in supplemented culture medium. This indicates that the culture medium influences the maintenance and development of the cultivated cells. The expression levels of and (related to the differentiation and migration of PGCs) were decreased in PGCs from the supplemented culture medium, supporting the results of ectopic migration. This is the first study to report the transplantation of PGCs to obtain germline chimera in neotropical species. The establishment of micromanipulation procedures in a model neotropical species will open new insights for the conservation of endangered species.
生物技术，包括源自原始生殖细胞（PGC）移植的代孕繁殖，是重建濒危鱼类物种的有价值的工具。尽管有前途，但以前没有使用新热带鱼类物种的此类方法的研究报告。本研究的目的是通过使用黄尾鲷作为鲇形目鱼类的模式物种来建立新热带鱼类的生殖系嵌合体。在不同条件下培养的 PGC 移植后获得了生殖系嵌合体：盐介质和补充有 DMEM、氨基酸、维生素、谷氨酰胺、丙酮酸盐和胎牛血清，随后将其移植到（♀）和（♂）杂交产生的三倍体和三倍体杂种中。结果表明，在盐介质中培养的 PGC 移植后，在宿主胚胎中发生异位迁移。然而，在补充培养基中培养的 PGC 迁移到了性腺嵴区域，在三倍体中为 4.5%，在三倍体杂种中为 19.3%。此外，在补充培养的培养基中培养的 PGC 中发现了 和 基因的高表达。这表明培养基影响培养细胞的维持和发育。在补充培养基中培养的 PGCs 的 和 （与 PGCs 的分化和迁移有关）的表达水平降低，支持了异位迁移的结果。这是首次报道在新热带物种中移植 PGC 以获得生殖系嵌合体的研究。在新热带模式物种中建立微操作程序将为濒危物种的保护开辟新的视野。
