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Preserving and Using Germplasm and Dissociated Embryonic Cells for Conserving Caribbean and Pacific Coral

机译:保存和利用种质和离体胚胎细胞保存加勒比和太平洋珊瑚

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摘要

Coral reefs are experiencing unprecedented degradation due to human activities, and protecting specific reef habitats may not stop this decline, because the most serious threats are global (i.e., climate change), not local. However, ex situ preservation practices can provide safeguards for coral reef conservation. Specifically, modern advances in cryobiology and genome banking could secure existing species and genetic diversity until genotypes can be introduced into rehabilitated habitats. We assessed the feasibility of recovering viable sperm and embryonic cells post-thaw from two coral species, Acropora palmata and Fungia scutaria that have diffferent evolutionary histories, ecological niches and reproductive strategies. In vitro fertilization (IVF) of conspecific eggs using fresh (control) spermatozoa revealed high levels of fertilization (>90% in A. palmata; >84% in F. scutaria; P>0.05) that were unaffected by tested sperm concentrations. A solution of 10% dimethyl sulfoxide (DMSO) at cooling rates of 20 to 30°C/min most successfully cryopreserved both A. palmata and F. scutaria spermatozoa and allowed producing developing larvae in vitro. IVF success under these conditions was 65% in A. palmata and 53% in F. scutaria on particular nights; however, on subsequent nights, the same process resulted in little or no IVF success. Thus, the window for optimal freezing of high quality spermatozoa was short (∼5 h for one night each spawning cycle). Additionally, cryopreserved F. scutaria embryonic cells had∼50% post-thaw viability as measured by intact membranes. Thus, despite some differences between species, coral spermatozoa and embryonic cells are viable after low temperature (−196°C) storage, preservation and thawing. Based on these results, we have begun systematically banking coral spermatozoa and embryonic cells on a large-scale as a support approach for preserving existing bio- and genetic diversity found in reef systems.
机译:由于人类活动,珊瑚礁正经历着前所未有的退化,保护特定的珊瑚礁栖息地可能无法阻止这种下降,因为最严重的威胁是全球性的(即气候变化),而不是局部性的。但是,非原生境保存做法可以为珊瑚礁的保存提供保障。特别是,冷冻生物学和基因组库的现代发展可以确保现有物种和遗传多样性,直到将基因型引入到经过修复的栖息地中。我们评估了从具有不同进化历史,生态位和繁殖策略的两种珊瑚物种,即鹿角Ac(Acropora palmata)和Fun木(Fungia scutaria)融化后恢复活精子和胚胎细胞的可行性。使用新鲜(对照)精子对同种卵的体外受精(IVF)显示受试精子浓度不受高水平的受精(在棕榈中> 90%;在中> 84%; P> 0.05)。 10%的二甲基亚砜(DMSO)溶液以20到30°C / min的冷却速度最成功地冷冻保存了棕榈曲霉和盾形F子精子,并在体外产生了发育中的幼虫。在特定条件下,在这些条件下,试管婴儿的试管婴儿成功率为65%,棕榈A为53%。但是,在随后的夜晚,相同的过程导致IVF成功很少或没有成功。因此,高质量精子最佳冷冻的窗口很短(每个产卵周期约一小时约5小时)。另外,如通过完整膜所测量的,冷冻保存的盾形葡萄球菌胚胎细胞具有约50%的解冻后活力。因此,尽管物种之间存在一些差异,但在低温(-196°C)储存,保存和解冻后,珊瑚精子和胚胎细胞仍然可行。基于这些结果,我们已开始系统地大规模保存珊瑚精子和胚胎细胞,以作为保护暗礁系统中现有生物和遗传多样性的一种支持方法。

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