de Souza Aline F, de Ramos Eloise C, Cury Fabio S, Pieri Naira Caroline G, Martins Daniele S
Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, Pirassununga, Brazil.
Department of Reproduction, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo, Brazil.
Reprod Domest Anim. 2019 Jul;54(7):964-971. doi: 10.1111/rda.13444. Epub 2019 May 18.
During the sex differentiation, the primordial germ cells (PGCs) pass through a differentiation, becoming spermatogonial cells in males and oocytes in females. In this phase, there is difference in gene expression and differentiation potency between males and females. Specific cell markers have been essential in the PGC meiosis beginning and become oocyte cells. However, there are few studies about germline in domestic animals. The domestic dog (Canis lupus familiaris) is an interesting animal model to be used in the investigation about the mammal development because it has several biochemical and physiological similarities to humans. In addition, some additional investigations about dogs may contribute to a better understanding of the biology and genetic components, improving clinical veterinary and zoological sciences. Here, we elucidated by immunofluorescence and quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), the dynamics of the expression of pluripotent (POU5F1 and NANOG) and germline (DDX4, DAZL and DPPA3) markers that are very important in the development of female canine germ cells during 35-50 days post-fertilization (dpf). The female canine germ cells were positive for pluripotent markers during middle developmental period. The number of DDX4, DAZL and DPPA3 cells increased along the germ cell maturation from 45 to 50 dpf. We provided an expression analysis of the pluripotent and germline markers in paraffin sections using the middle and later periods in female canine germ cells. The results can contribute the understanding about the timeline of each marker along the maturation of female canine germ cells. These results have a great significance to demonstrate the germ cell profile changes because it may allow the development of protocols about in vitro germ cell derivation.
在性别分化过程中,原始生殖细胞(PGCs)经历分化,在雄性中成为精原细胞,在雌性中成为卵母细胞。在此阶段,雄性和雌性之间存在基因表达和分化潜能的差异。特定的细胞标志物对于PGC减数分裂的开始以及成为卵母细胞至关重要。然而,关于家畜生殖系的研究很少。家犬(Canis lupus familiaris)是一种有趣的动物模型,可用于哺乳动物发育的研究,因为它在生物化学和生理学方面与人类有一些相似之处。此外,对狗的一些额外研究可能有助于更好地理解生物学和遗传成分,改善临床兽医学和动物学科学。在这里,我们通过免疫荧光和定量逆转录聚合酶链反应(RT-qPCR),阐明了多能性(POU5F1和NANOG)和生殖系(DDX4、DAZL和DPPA3)标志物在受精后35-50天(dpf)雌性犬生殖细胞发育过程中的表达动态。雌性犬生殖细胞在发育中期对多能性标志物呈阳性。从45到50 dpf,随着生殖细胞成熟,DDX4、DAZL和DPPA3细胞的数量增加。我们对雌性犬生殖细胞中期和后期石蜡切片中的多能性和生殖系标志物进行了表达分析。这些结果有助于了解每个标志物在雌性犬生殖细胞成熟过程中的时间线。这些结果对于证明生殖细胞谱的变化具有重要意义,因为它可能有助于制定体外生殖细胞衍生的方案。