An adaptable chromosome preparation methodology for use in invertebrate research organisms.
An adaptable chromosome preparation methodology for use in invertebrate research organisms.
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DOI：
10.1186/s12915-018-0497-4
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发表时间：
2018-02-26
期刊：
影响因子：
5.4
通讯作者：
Sánchez Alvarado A
中科院分区：
文献类型：
--
作者：
Guo L;Accorsi A;He S;Guerrero-Hernández C;Sivagnanam S;McKinney S;Gibson M;Sánchez Alvarado A
有效地可视化和操作染色体的能力是理解生物体基因组结构的基础。为哺乳动物细胞开发的传统染色体制备方法和依赖于物种特异性条件的方法不适用于许多无脊椎动物。因此，需要一种简单、廉价、适用于多种无脊椎动物物种的染色体制备方案。我们优化了染色体制备方案，并将其应用于几种涡虫物种（扁豆门）、淡水苹果蜗牛Pomacea canaliculata（软体动物门）和海葵Nematostella vectensis（刺胞动物门）。我们证明有丝分裂活性的成年组织和胚胎都可以作为中期染色体的来源，扩大了该技术在缺乏细胞系和/或无法获得完整生命周期的无脊椎动物中的潜在应用。用去离子水进行简单的低渗处理就足以进行核型；没有必要在培养基中培养细胞。获得的核型允许在其他形态上难以区分的生物体之间识别倍性和染色体结构的差异，例如在野外收集的混合涡虫种群。此外，我们表明，在代表三个不同门的所有测试生物中，该方案可以有效地与下游应用相结合，如染色体荧光原位杂交。我们的简单和廉价的染色体制备方案可以很容易地适用于新的无脊椎研究生物，以加速在生命之树的分支中发现新的基因组模式。
The ability to efficiently visualize and manipulate chromosomes is fundamental to understanding the genome architecture of organisms. Conventional chromosome preparation protocols developed for mammalian cells and those relying on species-specific conditions are not suitable for many invertebrates. Hence, a simple and inexpensive chromosome preparation protocol, adaptable to multiple invertebrate species, is needed. We optimized a chromosome preparation protocol and applied it to several planarian species (phylum Platyhelminthes), the freshwater apple snail Pomacea canaliculata (phylum Mollusca), and the starlet sea anemone Nematostella vectensis (phylum Cnidaria). We demonstrated that both mitotically active adult tissues and embryos can be used as sources of metaphase chromosomes, expanding the potential use of this technique to invertebrates lacking cell lines and/or with limited access to the complete life cycle. Simple hypotonic treatment with deionized water was sufficient for karyotyping; growing cells in culture was not necessary. The obtained karyotypes allowed the identification of differences in ploidy and chromosome architecture among otherwise morphologically indistinguishable organisms, as in the case of a mixed population of planarians collected in the wild. Furthermore, we showed that in all tested organisms representing three different phyla this protocol could be effectively coupled with downstream applications, such as chromosome fluorescent in situ hybridization. Our simple and inexpensive chromosome preparation protocol can be readily adapted to new invertebrate research organisms to accelerate the discovery of novel genomic patterns across the branches of the tree of life.
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影响因子：
2.4
作者：
Fritzenwanker, JH;Technau, U
通讯作者：
Technau, U
DOI：
10.2307/1352679
发表时间：
1994-06-01
期刊：
ESTUARIES
影响因子：
--
作者：
HAND, C;UHLINGER, KR
通讯作者：
UHLINGER, KR
影响因子：
1
作者：
Anokhin, B.;Hemmrich-Stanisak, G.;Bosch, T. C. G.
通讯作者：
Bosch, T. C. G.
影响因子：
4.6
作者：
Dorighi, Kristel M.;Tamkun, John W.
通讯作者：
Tamkun, John W.
影响因子：
--
作者：
Elliott, Sarah A.;Alvarado, Alejandro Sanchez
通讯作者：
Alvarado, Alejandro Sanchez