Transcriptomics of Arabidopsis sperm cells at single-cell resolution
Transcriptomics of Arabidopsis sperm cells at single-cell resolution
复制标题
DOI：
10.1007/s00497-018-00355-4
复制
发表时间：
2019-03-01
影响因子：
3.4
通讯作者：
Becker, Joerg D.
中科院分区：
文献类型：
--
作者：
Misra, Chandra Shekhar;Santos, Mario R.;Becker, Joerg D.
关键信息我们提出了分离单个精子细胞和转录组分析的详细方案，以研究精子细胞之间基因表达的变异。 摘要开花植物中的雄性配子体发育始于小孢子母细胞，在连续两次细胞分裂后形成包含营养核和两个精子细胞的成熟花粉粒。花粉发育是一个高度动态的过程，涉及营养核和一对具有自己的细胞质和细胞核的精子细胞的转录组和表观基因组水平的变化。虽然拟南芥花粉发育的整体转录组已有详细记录，但单细胞水平的研究，特别是精子细胞的研究，仍然缺乏。这些研究对于了解两个精子细胞在转录上是否存在差异以及如何存在差异至关重要，特别是当花粉管通过雌蕊的传递组织生长时。在这里，我们描述了从生长的花粉管中分离单个精子细胞并分析其转录组的详细方案。
Key messageWe present a detailed protocol for isolation of single sperm cells and transcriptome analysis to study variation in gene expression between sperm cells.AbstractMale gametophyte development in flowering plants begins with a microspore mother cell, which upon two consecutive cell divisions forms a mature pollen grain containing a vegetative nucleus and two sperm cells. Pollen development is a highly dynamic process, involving changes at both the transcriptome and epigenome levels of vegetative nuclei and the pair of sperm cells that have their own cytoplasm and nucleus. While the overall transcriptome of Arabidopsis pollen development is well documented, studies at single-cell level, in particular of sperm cells, are still lacking. Such studies would be essential to understand whether and how the two sperm cells are transcriptionally different, in particular once the pollen tube grows through the transmitting tissue of the pistil. Here we describe a detailed protocol for isolation of single sperm cells from growing pollen tubes and analysis of their transcriptome.