Doctoral Degree Program in Marine Biotechnology, National Taiwan Ocean University, Keelung, Taiwan.
Doctoral Degree Program in Marine Biotechnology, Academia Sinica, Taipei, Taiwan.
Mol Reprod Dev. 2019 Jul;86(7):798-811. doi: 10.1002/mrd.23157. Epub 2019 May 6.
In a variety of organisms, adult gonads contain several specialized somatic cells that regulate and support the development of germline cells. In stony corals, the characteristics and functions of gonadal somatic cells remain largely unknown. No molecular markers are currently available that allow for the identification and enrichment of gonadal somatic cells in corals. Here, we showed that the testicular somatic cells of a stony coral, Euphyllia ancora, express an endogenous green fluorescent protein (GFP). Fluorescence microscopy showed that, in contrast to the endogenous expression of the red fluorescent protein of E. ancora ovaries that we have previously reported, the testes displayed a distinct green fluorescence. Molecular identification and spectrum characterization demonstrated that E. ancora testes expressed a GFP (named EaGFP) that is a homolog of the GFP from the jellyfish Aequorea victoria and that possesses an excitation maximum of 506 nm and an emission maximum of 514 nm. Immunohistochemical analyses revealed that the testicular somatic cells, but not the germ cells, expressed EaGFP. EaGFP was enclosed within one or a few granules in the cytoplasm of testicular somatic cells, and the granule number decreased as spermatogenesis proceeded. We also showed that testicular somatic cells could be enriched by using endogenous GFP as an indicator. The present study not only revealed one of the unique cellular characteristics of coral testicular cells but also established a technical basis for more in-depth investigations of the function of testicular somatic cells in spermatogenesis in future studies.
在多种生物体中，成年性腺包含几种专门的体细胞，这些体细胞调节和支持生殖细胞的发育。在石珊瑚中，性腺体细胞的特征和功能在很大程度上仍不清楚。目前没有分子标记物可以用于鉴定和富集珊瑚中的性腺体细胞。在这里，我们表明，一种石珊瑚(Euphyllia ancora)的睾丸体细胞表达内源性绿色荧光蛋白(GFP)。荧光显微镜显示，与我们之前报道的 E. ancora 卵巢中红色荧光蛋白的内源性表达相比，睾丸显示出明显的绿色荧光。分子鉴定和光谱特征表明，E. ancora 睾丸表达了一种 GFP(EaGFP)，它是水母 Aequorea victoria 的 GFP 的同源物，具有 506nm 的最大激发波长和 514nm 的最大发射波长。免疫组织化学分析显示，睾丸体细胞而不是生殖细胞表达 EaGFP。EaGFP 被包裹在睾丸体细胞细胞质中的一个或几个颗粒内，随着精子发生的进行，颗粒数量减少。我们还表明，可以使用内源性 GFP 作为指标来富集睾丸体细胞。本研究不仅揭示了珊瑚睾丸细胞的独特细胞特征之一，还为未来研究中更深入研究睾丸体细胞在精子发生中的功能奠定了技术基础。
