Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Aquaculture Genetics and Breeding Laboratory, Ave. Instituto Politécnico Nacional No. 195, Col. Playa Palo de Santa Rita, 23096 La Paz, Baja California Sur, Mexico.
Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Aquaculture Genetics and Breeding Laboratory, Ave. Instituto Politécnico Nacional No. 195, Col. Playa Palo de Santa Rita, 23096 La Paz, Baja California Sur, Mexico.
Gene. 2019 Jul 20;706:19-31. doi: 10.1016/j.gene.2019.04.066. Epub 2019 Apr 24.
The fem-1 gene in Caenorhabditis elegans is involved in sex differentiation; it is specifically required for all aspects of male development. In this study, the full-length cDNA of the fem-1 (Pvfem-1) gene was isolated from the Pacific whiteleg shrimp Penaeus vannamei. The Pvfem-1 transcript is 3778 nt long and encodes a putative protein (PvFEM-1) of 638 amino acids that presented eight ankyrin repeats. The translated protein showed a significant (P < 0.05) structural similitude by superposition with C. elegans FEM-1 protein. Pvfem-1 expression was evaluated by qPCR and in situ hybridization (ISH) during embryogenesis, larval development, and gonads of both genders in subadult and adult life stages. Pvfem-1 was found expressed in brain, intestine, hepatopancreas, and in the gonads of both genders in subadults and adults when quantified by RT-qPCR. A significant finding was the discovery of a natural antisense transcript (NAT) of Pvfem-1 by ISH. It was present in the oocyte nucleus of subadult female shrimp gonads but was not seen within oocytes from adult females, although it was detected in follicular cells, suggesting a possible post-transcriptional regulation of Pvfem-1 in female gonad. Conversely, in males, no NAT was observed, and Pvfem-1 was found expressed in spermatogonia of both, subadult and adult shrimps indicating a function in male sexual differentiation and gametes generation. This study represents the first step for future functional analysis that is expected to contribute to clarifying the role of Pvfem-1 in sex differentiation and determination.
秀丽隐杆线虫的 fem-1 基因参与性别分化；它是雄性发育所有方面所必需的。在这项研究中，从凡纳滨对虾（Penaeus vannamei）中分离出全长 fem-1（Pvfem-1）基因的 cDNA。Pvfem-1 转录本长 3778nt，编码一个假定的蛋白质（PvFEM-1），由 638 个氨基酸组成，具有 8 个锚蛋白重复序列。该翻译蛋白通过与秀丽隐杆线虫 FEM-1 蛋白的叠加显示出显著的（P < 0.05）结构相似性。通过 qPCR 和原位杂交（ISH）在胚胎发生、幼虫发育以及亚成体和成年阶段雌雄两性性腺中评估了 Pvfem-1 的表达。通过 RT-qPCR 定量发现，Pvfem-1 在亚成体和成年个体的大脑、肠道、肝胰腺以及两性性腺中表达。通过 ISH 发现了 Pvfem-1 的天然反义转录物（NAT），它存在于亚成体雌性虾性腺的卵母细胞核中，但在成年雌性的卵母细胞中未观察到，尽管在滤泡细胞中检测到，这表明 Pvfem-1 在雌性性腺中的转录后调控可能存在。相反，在雄性中未观察到 NAT，并且在亚成体和成年虾的精原细胞中发现 Pvfem-1 表达，表明它在雄性性分化和配子生成中具有功能。本研究代表了未来功能分析的第一步，有望有助于阐明 Pvfem-1 在性别分化和决定中的作用。
