Ricci Juliana M B, Martinez Emanuel R M, Butzge Arno J, Doretto Lucas B, Oliveira Marcos A, Bombardelli Robie Allan, Bogerd Jan, Nóbrega Rafael H
Reproductive and Molecular Biology Group, Department of Morphology, Institute of Bioscience of Botucatu, São Paulo State University, Botucatu, São Paulo, Brazil.
Center of Engineering and Exact Sciences, Universidade Estadual do Oeste do Paraná, Rua da Faculdade 645, 85903-000 Toledo, PR, Brazil.
Gene. 2018 May 15;654:116-126. doi: 10.1016/j.gene.2018.02.029. Epub 2018 Feb 14.
We have characterized the full-length vasa cDNA from Jundiá, Rhamdia quelen (Heptapteridae, Siluriformes). vasa encodes a member of the DEAD-box protein family of ATP-dependent RNA helicases. This protein is highly conserved among different organisms and its role is associated with RNA metabolism. In the majority of the investigated species, vasa is restricted to the germ cell lineage and its expression has been used to study germline development in many organisms, including fish. The deduced R. quelen vasa amino acid sequence displayed high similarity with Vasa protein sequences from other organisms, and did not cluster with PL10 or P68 DEAD-box protein subfamilies. We also reported that there is no other isoform for vasa mRNA in R. quelen gonads. Expression analysis by RT-PCR and qPCR showed vasa transcripts exclusively expressed in the germ cells of R. quelen gonads. R. quelen vasa mRNA was maternally inherited, and was detected in the migrating primordial germ cells (PGCs) until 264 h post-fertilization during embryonic and larval development. This work has characterized for the first time the full-length R. quelen vasa cDNA, and describes its expression patterns during R. quelen embryonic and larval development. Our results will contribute to the basic reproductive biology of this native species, and will support studies using vasa as a germ cell marker in different biotechnological studies, such as germ cell transplantation.
我们已经对来自胡鲶(Rhamdia quelen,海鲶科,鲶形目)的全长瓦萨(vasa)cDNA进行了特征分析。瓦萨编码一种依赖ATP的RNA解旋酶的DEAD盒蛋白家族成员。这种蛋白质在不同生物中高度保守,其作用与RNA代谢相关。在大多数被研究的物种中,瓦萨仅限于生殖细胞谱系,其表达已被用于研究包括鱼类在内的许多生物的生殖系发育。推导的胡鲶瓦萨氨基酸序列与其他生物的瓦萨蛋白序列显示出高度相似性,并且不与PL10或P68 DEAD盒蛋白亚家族聚类。我们还报道了在胡鲶性腺中瓦萨mRNA没有其他异构体。通过RT-PCR和qPCR进行的表达分析表明,瓦萨转录本仅在胡鲶性腺的生殖细胞中表达。胡鲶瓦萨mRNA是母系遗传的,并且在胚胎和幼体发育期间,在受精后264小时之前的迁移原始生殖细胞(PGC)中被检测到。这项工作首次对全长胡鲶瓦萨cDNA进行了特征分析,并描述了其在胡鲶胚胎和幼体发育期间的表达模式。我们的结果将有助于该本地物种的基础生殖生物学研究,并将支持在不同生物技术研究(如生殖细胞移植)中使用瓦萨作为生殖细胞标记的研究。