Microscopy of Physcomitrella patens sperm cells.
Microscopy of Physcomitrella patens sperm cells.
复制标题
DOI：
10.1186/s13007-017-0186-2
复制
发表时间：
2017
期刊：
影响因子：
5.1
通讯作者：
Reski R
中科院分区：
文献类型：
--
作者：
Horst NA;Reski R
脊椎动物(苔藓、蕨类和裸子植物)，如苔藓，拥有自由活动的精子细胞(精子)，通过地表水到达卵细胞。虽然这些可移动的鞭毛精子细胞是一个传统的植物学主题，但它们在旗舰非种子植物模式物种P.patens中还没有得到彻底的分析。需要通过协议来确定野生型精子的行为，作为未来研究的先决条件，例如确定影响精子数量、形态、活力和活力的突变或因素的特征。在这里，我们介绍了使用标准显微镜以中等放大倍数观察固定精子和活精子的方案。固定样本可用于精子数量和形态的快速评估。为了确定功能，需要观察活精子。提供了确定精子活力和存活率的方案，允许区分这两个参数。这些循序渐进的方案对于到目前为止还不熟悉活动配子分析的研究人员特别有用，旨在帮助建立和改进这些分析，以便促进苔藓模式物种P.patens精子发生的研究。
Archegoniates (bryophytes, ferns and gymnosperms), such as the moss Physcomitrella patens, possess freely motile sperm cells (spermatozoids) which reach the egg cell via surface water. Although these motile flagellated sperm cells are a traditional botanical subject, they have not been thoroughly analysed in the flagship non-seed plant model species P. patens. Protocols are required to determine the behaviour of wild type sperms as a prerequisite for future research such as the characterization of mutants or factors that influence sperm number, morphology, viability and motility. Here, we present protocols for the observation of fixed, as well as live sperms utilizing a standard microscope at intermediate magnifications. Fixed samples can be used for the fast assessment of sperm number and morphology. To determine functionality, the observation of live sperms is required. Protocols for determining both sperm motility and viability are provided, allowing both parameters to be distinguished. These step-by-step protocols are particularly useful for researchers so far not familiar with the analysis of motile gametes and are meant to aid the establishment and improvement of these analyses in order to stimulate research on spermatogenesis in the moss model species P. patens.