López-Galindo Laura, Juárez Oscar E, Larios-Soriano Ernesto, Del Vecchio Giulia, Ventura-López Claudia, Lago-Lestón Asunción, Galindo-Sánchez Clara
Laboratorio de Genómica Funcional, Departamento de Biotecnología Marina, Centro de Investigación Científica y de Educación Superior de Ensenada, Ensenada, Mexico.
Laboratorio de Fisiología Integrativa de Organismos Marinos, Departamento de Biotecnología Marina, Centro de Investigación Científica y de Educación Superior de Ensenada, Ensenada, Mexico.
Front Physiol. 2019 Jan 15;9:1920. doi: 10.3389/fphys.2018.01920. eCollection 2018.
endemic to the Yucatan Peninsula, Mexico, is an ectotherm organism particularly temperature-sensitive. Studies in females show that temperatures above 27°C reduce the number of eggs per spawn, fertilization rate and the viability of embryos. High temperatures also reduce the male reproductive performance and success. However, the molecular mechanisms are still unknown. The transcriptomic profiles of testes from thermally stressed (30°C) and not stressed (24°C) adult male octopuses were compared, before and after mating to understand the molecular bases involved in the low reproductive performance at high temperature. The testis paired-end cDNA libraries were sequenced using the Illumina MiSeq platform. Then, the transcriptome was assembled using Trinity software. A total of 53,214,611 high-quality paired reads were used to reconstruct 85,249 transcripts and 77,661 unigenes with an N50 of 889 bp length. Later, 13,154 transcripts were annotated implementing Blastx searches in the UniProt database. Differential expression analysis revealed 1,881 transcripts with significant difference among treatments. Functional annotation and pathway mapping of differential expressed transcripts revealed significant enrichment for biological processes involved in spermatogenesis, gamete generation, germ cell development, spermatid development and differentiation, response to stress, inflammatory response and apoptosis. Remarkably, the transcripts encoding genes such as ZMYND15, KLHL10, TDRD1, TSSK2 and DNAJB13, which are linked to male infertility in other species, were differentially expressed among the treatments. The expression levels of these key genes, involved in sperm motility and spermatogenesis were validated by quantitative real-time PCR. The results suggest that the reduction in male fertility at high temperature can be related to alterations in spermatozoa development and motility.
墨西哥尤卡坦半岛特有的一种变温生物对温度特别敏感。对雌性的研究表明,高于27°C的温度会减少每次产卵的数量、受精率和胚胎的活力。高温还会降低雄性的生殖性能和成功率。然而,分子机制仍然未知。比较了热应激(30°C)和未应激(24°C)成年雄性章鱼交配前后睾丸的转录组图谱,以了解高温下生殖性能低下所涉及的分子基础。使用Illumina MiSeq平台对睾丸双末端cDNA文库进行测序。然后,使用Trinity软件组装转录组。总共53,214,611条高质量双末端读段用于重建85,249个转录本和77,661个单基因,N50长度为889 bp。随后,通过在UniProt数据库中进行Blastx搜索对13,154个转录本进行注释。差异表达分析显示,各处理之间有1,881个转录本存在显著差异。差异表达转录本的功能注释和通路映射显示,参与精子发生、配子生成、生殖细胞发育、精子细胞发育和分化、应激反应、炎症反应和细胞凋亡的生物学过程显著富集。值得注意的是,在其他物种中与男性不育相关的ZMYND15、KLHL10、TDRD1、TSSK2和DNAJB13等基因的转录本在各处理之间差异表达。通过定量实时PCR验证了这些参与精子活力和精子发生的关键基因的表达水平。结果表明,高温下雄性生育力的降低可能与精子发育和活力的改变有关。