Dipartimento di Scienze della Terra e del Mare, Viale delle Scienze Ed. 16, Palermo, Italy.
Dipartimento di Scienze e Tecnologie Biologiche, Chimiche e Farmaceutiche, Via Archirafi 18, Palermo, Italy.
Fish Shellfish Immunol. 2018 Jan;72:452-458. doi: 10.1016/j.fsi.2017.11.023. Epub 2017 Nov 13.
Riboprobes (in situ hybridization) and antibodies (immunohistochemistry), previously used to show the upregulation of Ciona intestinalis (Type A) galectins (CiLgals-a, CiLgals-b) and phenoloxidase (CinPO2) immune-related genes, were tested on histological sections of the ovary. The ovarian follicles are composed of oocytes encased by follicular cells (FCs) and test cells (TCs). Results show the transcription upregulation of both CiLgals and CinPO2 genes in the vitellogenic FCs, conversely distinct cytolocalization of the proteins are shown. At vitellogenic stage, the CiLgals are localized in the FCs, in the oocyte cytoplasm, and close to the germinal vesicle (GV), whereas the CinPO2 was never identified in the FCs. In a presumptive advanced phase and at the post-vitellogenic stage the TCs appear to be labelled by the CinPO2 riboprobe, and the protein identified by the antibody suggesting an mRNA transcytosis process from FCs. At post-vitellogenic stage the CiLgals mainly enrich the GV nucleoplasm, whereas the CinPO2 is contained in TCs and in the ooplasm but never found in the GV. This finding sheds new light on a former paper in which TCs were reported to be the only CinPO2-producing cells in the ovarian follicle. Finally, CiLgals and CinPO2 genes transcription and proteins production seem to be associated with accessory cells during their differentiation from vitellogenic to post-vitellogenic stage. The present findings promote further research on the early upregulation of immune-related genes, and the potential multifunctional role of the produced proteins. In addition further insight on the accessory cells involvement in ascidian oogenesis are reported.
先前使用核糖探针（原位杂交）和抗体（免疫组织化学）来显示文昌鱼（A 型）半乳糖凝集素（CiLgals-a、CiLgals-b）和酚氧化酶（CinPO2）免疫相关基因的上调，现已在卵巢组织切片上进行了测试。卵巢滤泡由被滤泡细胞（FCs）和滋养细胞（TCs）包围的卵母细胞组成。结果表明，在卵黄生成的 FCs 中，CiLgals 和 CinPO2 基因的转录均上调，而蛋白质的细胞定位则明显不同。在卵黄生成阶段，CiLgals 定位于 FCs、卵母细胞质和接近生殖泡（GV），而 CinPO2 从未在 FCs 中被鉴定出。在假定的晚期和卵黄生成后阶段，TCs 似乎被 CinPO2 核糖探针标记，并用抗体鉴定出的蛋白质表明存在从 FCs 中转录物胞内转运的过程。在卵黄生成后阶段，CiLgals 主要富集在 GV 核质中，而 CinPO2 则存在于 TCs 和卵质中，但从未在 GV 中发现。这一发现为以前的一篇论文提供了新的线索，该论文报道 TCs 是卵巢滤泡中唯一产生 CinPO2 的细胞。最后，CiLgals 和 CinPO2 基因的转录和蛋白质的产生似乎与附属细胞在从卵黄生成到卵黄生成后的分化过程中有关。本研究结果促进了对免疫相关基因早期上调以及产生的蛋白质的潜在多功能作用的进一步研究。此外，还报道了关于附属细胞参与海鞘卵子发生的更多见解。
