Initial response of ovarian tissue transcriptome to vitrification or microwave-assisted dehydration in the domestic cat model.
Initial response of ovarian tissue transcriptome to vitrification or microwave-assisted dehydration in the domestic cat model.
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DOI：
10.1186/s12864-020-07236-z
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发表时间：
2020-11-25
期刊：
影响因子：
4.4
通讯作者：
Comizzoli P
中科院分区：
文献类型：
--
作者：
Amelkina O;Comizzoli P
卵巢组织的长期保存是人类生殖医学和珍稀动物基因型保护的重要手段。与单细胞保存相比，组织方案的优化是高度复杂的，因为细胞的多样性对非生理条件的反应不同。以青春期前家猫为模型，研究玻璃化冷冻或微波辅助脱水对卵巢皮质整体转录组动态的即时影响。对来自不同条件的卵巢组织（n = 6个个体）进行RNA测序：解剖后的新鲜组织（F）、玻璃化/温热组织（V）、脱水5 min（D5）或10 min（D10）然后再水化的组织。对比较对V vs. F、D10 vs. F、D5 vs. F和D10 vs. D5进行差异基因表达分析，并基于功能富集和计算机模拟蛋白质-蛋白质相互作用的结果构建网络。玻璃化冷冻方案的影响在升温后20分钟内已经可以测量，并且涉及与线粒体呼吸相关的7个线粒体DNA基因表达的上调。D10与F的分析显示，再水化后30分钟，基因表达的主要下调，Ras、Rap 1、PI 3 K-Akt和MAPK信号通路中的计算机相互作用基因富集。然而，D5与F的比较显示了具有两个富含Ras信号传导的基因的较短脱水方案的可忽略的影响。D10与D5的比较显示仅7个基因下调。玻璃化和脱水方案主要改变了不同基因和功能术语的表达，但一些差异表达的基因形成了一个主要的计算机蛋白质-蛋白质相互作用簇，富集线粒体呼吸和Ras/MAPK信号通路。我们的研究结果表明，第一次，不同的影响玻璃化和微波辅助脱水协议的全球转录组的卵巢皮质（使用家猫作为生物医学模型）。基于差异表达基因构建的数据和网络（1）可以帮助更好地理解对非生理应激的应激反应，（2）可以用作未来保存方案优化的方向。在线版本包含补充材料，可通过10.1186/s12864-020-07236-z获得。
Long term preservation of living ovarian tissues is a critical approach in human reproductive medicine as well as in the conservation of rare animal genotypes. Compared to single cell preservation, optimization of protocols for tissues is highly complex because of the diversity of cells responding differently to non-physiological conditions. Using the prepubertal domestic cat as a model, the objective was to study immediate effects of vitrification or microwave-assisted dehydration on the global transcriptome dynamics in the ovarian cortex. RNA sequencing was performed on ovarian tissues (n = 6 individuals) from different conditions: fresh tissue after dissection (F), vitrified/warmed tissue (V), tissue dehydrated for 5 min (D5) or 10 min (D10) followed by rehydration. Differential gene expression analysis was performed for comparison pairs V vs. F, D10 vs. F, D5 vs. F and D10 vs. D5, and networks were built based on results of functional enrichment and in silico protein-protein interactions. The impact of the vitrification protocol was already measurable within 20 min after warming and involved upregulation of the expression of seven mitochondrial DNA genes related to mitochondrial respiration. The analysis of D10 vs. F revealed, 30 min after rehydration, major downregulation of gene expression with enrichment of in silico interacting genes in Ras, Rap1, PI3K-Akt and MAPK signaling pathways. However, comparison of D5 vs. F showed negligible effects of the shorter dehydration protocol with two genes enriched in Ras signaling. Comparison of D10 vs. D5 showed downregulation of only seven genes. Vitrification and dehydration protocols mainly changed the expression of different genes and functional terms, but some of the differentially expressed genes formed a major in silico protein-protein interaction cluster enriched for mitochondrial respiration and Ras/MAPK signaling pathways. Our results showed, for the first time, different effects of vitrification and microwave-assisted dehydration protocols on the global transcriptome of the ovarian cortex (using the domestic cat as a biomedical model). Acquired data and networks built on the basis of differentially expressed genes (1) can help to better understand stress responses to non-physiological stresses and (2) can be used as directions for future preservation protocol optimizations. The online version contains supplementary material available at 10.1186/s12864-020-07236-z.
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影响因子：
1.6
作者：
Elliott, Gloria D.;Lee, Pei-Chih;Comizzoli, Pierre
通讯作者：
Comizzoli, Pierre
影响因子：
1.6
作者：
Amoushahi, Mahboobeh;Salehnia, Mojdeh;Ghorbanmehr, Nassim
通讯作者：
Ghorbanmehr, Nassim
影响因子：
4.3
作者：
Hand SC;Menze MA
通讯作者：
Menze MA
DOI：
10.1073/pnas.0808480105
发表时间：
2008-10-14
影响因子：
11.1
作者：
Jao, Cindy Y.;Salic, Adrian
通讯作者：
Salic, Adrian
DOI：
10.1016/s1095-6433(01)00503-7
发表时间：
2002-03-01
影响因子：
2.3
作者：
Crowe, LM
通讯作者：
Crowe, LM