Genome Editing Innovation Center, Hiroshima University, 3-10-23 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-0046, Japan.
Graduate School of Integrated Sciences for Life, Hiroshima University, 1-4-4 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8528, Japan.
Sci Rep. 2022 Aug 17;12(1):13528. doi: 10.1038/s41598-022-17726-7.
In birds, sex is determined through cell-autonomous mechanisms and various factors, such as the dosage of DMRT1. While the sex-determination mechanism in gonads is well known, the mechanism in germ cells remains unclear. In this study, we explored the gene expression profiles of male and female primordial germ cells (PGCs) during embryogenesis in chickens to predict the mechanism underlying sex determination. Male and female PGCs were isolated from blood and gonads with a purity > 96% using flow cytometry and analyzed using RNA-seq. Prior to settlement in the gonads, female circulating PGCs (cPGCs) obtained from blood displayed sex-biased expression. Gonadal PGCs (gPGCs) also exhibited sex-biased expression, and the number of female-biased genes detected was higher than that of male-biased genes. The female-biased genes in gPGCs were enriched in some metabolic processes. To reveal the mechanisms underlying the transcriptional regulation of female-biased genes in gPGCs, we performed stimulation tests. Retinoic acid stimulation of cultured gPGCs derived from male embryos resulted in the upregulation of several female-biased genes. Overall, our results suggest that sex determination in avian PGCs involves aspects of both cell-autonomous and somatic-cell regulation. Moreover, it appears that sex determination occurs earlier in females than in males.
在鸟类中，性别是通过细胞自主机制和各种因素决定的，例如 DMRT1 的剂量。虽然性腺的性别决定机制已经很清楚，但生殖细胞中的机制仍不清楚。在这项研究中，我们通过流式细胞术从血液和性腺中分离出纯度大于 96%的雄性和雌性原始生殖细胞（PGC），并使用 RNA-seq 进行分析，以探索鸡胚胎发生过程中雄性和雌性 PGC 的基因表达谱，从而预测性别决定的机制。在定居在性腺之前，来自血液的雌性循环 PGC（cPGC）表现出性别偏倚表达。性腺 PGC（gPGC）也表现出性别偏倚表达，并且检测到的雌性偏倚基因数量高于雄性偏倚基因。gPGC 中的雌性偏倚基因富集在一些代谢过程中。为了揭示 gPGC 中雌性偏倚基因转录调控的机制，我们进行了刺激试验。对来自雄性胚胎的培养 gPGC 进行维甲酸刺激导致几个雌性偏倚基因的上调。总的来说，我们的结果表明，禽类 PGC 的性别决定涉及细胞自主和体细胞调节的方面。此外，雌性的性别决定似乎比雄性更早发生。
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