Department of Biological, Geological and Environmental Sciences, BiGeA, University of Bologna, Bologna, Italy.
Histochem Cell Biol. 2021 Jul;156(1):19-34. doi: 10.1007/s00418-021-01983-0. Epub 2021 Mar 26.
The germline is a key feature of sexual animals and the ways in which it separates from the soma differ widely across Metazoa. However, at least at some point during germline differentiation, some cytoplasmic supramolecular structures (collectively called germ plasm-related structures) are present and involved in its specification and/or differentiation. The factors involved in the assembly of these granular structures are various and non-ubiquitous among animals, even if some functional patterns and the presence of certain domains appear to be shared among some. For instance, the LOTUS domain is shared by Oskar, the Holometabola germ plasm master regulator, and some Tudor-family proteins assessed as being involved in the proper assembly of germ granules of different animals. Here, we looked for the presence of LOTUS-containing proteins in the transcriptome of Ruditapes philippinarum (Bivalvia). Such species is of particular interest because it displays annual renewal of gonads, sided by the renewal of germline differentiation pathways. Moreover, previous works have identified in its early germ cells cytoplasmic granules containing germline determinants. We selected the orthologue of TDRD7 as a candidate involved in the early steps of germline differentiation through bioinformatic predictions and immunohistological patterning (immunohistochemistry and immunofluorescence). We observed the expression of the protein in putative precursors of germline cells, upstream to the germline marker Vasa. This, added to the fact that orthologues of this protein are involved in the assembly of germ granules in mouse, zebrafish, and fly, makes it a worthy study unit for investigations on the formation of such structures in bivalves.
生殖系是有性动物的一个关键特征,其与体细胞分离的方式在后生动物中差异很大。然而,至少在生殖系分化的某个阶段,一些细胞质超分子结构(统称为生殖质相关结构)存在并参与其特化和/或分化。参与这些颗粒状结构组装的因素多种多样,在动物中并不普遍存在,即使某些功能模式和某些结构域的存在似乎在某些动物中共享。例如,LOTUS 结构域存在于 Oskar(完全变态类的生殖质主调控因子)和一些 Tudor 家族蛋白中,这些蛋白被评估为参与不同动物生殖颗粒的正确组装。在这里,我们在菲律宾蛤仔(双壳纲)的转录组中寻找含有 LOTUS 蛋白的存在。这种物种特别有趣,因为它的性腺每年都会更新,同时生殖系分化途径也会更新。此外,之前的工作已经在其早期生殖细胞中鉴定到含有生殖系决定因素的细胞质颗粒。我们通过生物信息学预测和免疫组织化学模式(免疫组织化学和免疫荧光)选择 TDRD7 的同源物作为参与生殖系分化早期步骤的候选物。我们观察到该蛋白在生殖系标记物 Vasa 之前的生殖系细胞前体中表达。此外,该蛋白的同源物参与了小鼠、斑马鱼和果蝇中生殖颗粒的组装,这使得它成为研究双壳类动物中这些结构形成的一个有价值的研究单元。