Abstract：
Primordial germ cells (PGCs) are embryonic pluripotent cells that can differentiate into spermatogonia and oogonia, and therefore, PGCs are a genetic source for germplasm conservation through cryobanking and the generation of germline chimeras. The knowledge of PGC migration routes is essential for transplantation studies. In this work, the mRNA synthesized from the ddx4 3\'UTR sequence of Pseudopimelodus mangurus, in fusion with gfp or dsred, was microinjected into zygotes of three neotropical species (P. mangurus, Astyanax altiparanae, and Prochilodus lineatus) for PGC labeling. Visualization of labeled PGCs was achieved by fluorescence microscopy during embryonic development. In addition, ddx4 and dnd1 expressions were evaluated during embryonic development, larvae, and adult tissues of P. mangurus, to validate their use as a PGC marker. As a result, the effective identification of presumptive PGCs was obtained. DsRed-positive PGC of P. mangurus was observed in the hatching stage, GFP-positive PGC of A. altiparanae in the gastrula stage, and GFP-positive PGCs from P. lineatus were identified at the segmentation stage, with representative labeling percentages of 29% and 16% in A. altiparanae and P. lineatus, respectively. The expression of ddx4 and dnd1 of P. mangurus confirmed the specificity of these genes in germ cells. These results point to the functionality of the P. mangurus ddx4 3\'UTR sequence as a PGC marker, demonstrating that PGC labeling was more efficient in A. altiparanae and P. lineatus. The procedures used to identify PGCs in P. mangurus consolidate the first step for generating germinal chimeras as a conservation action of P. mangurus.
摘要：
原始生殖细胞（PGCs）是胚胎多能细胞，可以分化为精原细胞和卵原细胞，因此,PGCs是通过冷冻保存和产生种系嵌合体进行种质保存的遗传来源。PGC迁移途径的知识对于移植研究至关重要。在这项工作中,从假山楂的ddx43UTR序列合成的mRNA，与gfp或dsred融合，被显微注射到三个新热带物种的受精卵中(P.芒果,Astyanaxaltiparanae，和Prochiloduslineatus）用于PGC标记。在胚胎发育期间通过荧光显微镜实现标记的PGCs的可视化。此外,在胚胎发育过程中评估ddx4和dnd1的表达，幼虫,和成虫组织，以验证它们作为PGC标记的用途。因此,获得了推定PGCs的有效鉴定。在孵化阶段观察到了DsRed阳性的PGC。原肠胚阶段的高原虫GFP阳性PGC，并且在分割阶段鉴定了来自直线疟原虫的GFP阳性PGCs，具有代表性的标签百分比分别为29%和16%。分别。mangurus的ddx4和dnd1的表达证实了这些基因在生殖细胞中的特异性。这些结果指出了作为PGC标记的马虎假单胞菌ddx43'UTR序列的功能，证明PGC标记在Altiparanae和Lineatus中更有效。用于鉴定马urus中的PGCs的程序巩固了产生生发嵌合体作为马urus的保护作用的第一步。
