Observation of polarity induction by cytochemical localization of phenylalkylamine-binding sites in regenerating protoplasts of the moss Physcomitrella patens
Observation of polarity induction by cytochemical localization of phenylalkylamine-binding sites in regenerating protoplasts of the moss Physcomitrella patens
复制标题
DOI：
10.1007/s007090200010
复制
发表时间：
2002-01-01
期刊：
影响因子：
2.9
通讯作者：
Reski, R
中科院分区：
文献类型：
--
作者：
Bhatla, SC;Kiessling, J;Reski, R
不同的外部光)和内部(例如生长素和钙梯度)因子控制苔藓原丝体的分化，目前的研究表明外源应用的生长素。萘基邻苯二甲酸对生长素外流的药理阻断作用。并用(-)苯丙地尔治疗。另一种钙通道拮抗剂苯烷胺(DM-BODIPY PAA)是一种钙通道拮抗剂，在不影响原生质体活力的情况下抑制原生质体的分裂。用LIS作为活体标记苯烷胺(PAA)结合位点的探针。这种结合的特异性通过与(-)苯丙地尔的竞争而得到证实。激光共聚焦扫描显微镜可见PAA结合部位在质膜和核膜上呈均匀分布的簇状。然而，在P Patens原生质体的不对称分裂过程中，荧光标记在膜内陷以及后来沿着分隔新细胞的平板，细胞内PAA结合部位的定位尤其增加。可能是在囊泡和液泡的膜上，在较小的子细胞中显著增加，注定要在以后形成一个极地突起。第一个绿藻细胞。因此，建立了一个在亚细胞水平上可视化早熟禾原生质体极化事件的系统。
Different external light) and internal (e.g.. auxin and calcium gradients) factors control differentiation of the moss protonema, The present investigations demonstrate that exogenously applied auxin. the pharmacological blockade of auxin efflux by naphthylphthalamic acid. and treatment with (-)bepridil. a calcium channel antagonist, inhibit protoplast division without affecting protoplast viability in the moss Physcomitrella patens, A fluorescently labelled phenylalkylamine (DM-Bodipy PAA), another calcium channel antagonist. was used Lis a probe for in vivo labelling of phenylalkylamine(PAA)-binding sites. The specificity of this binding was demonstrated by competition with (-)bepridil. Confocal laser scanning microscopy visualized PAA-binding sites on the plasma membrane and along the nuclear membrane as uniformly distributed clusters. During asymmetric division of P patens protoplasts, however, fluorescence labelling particularly increases at the membrane invagination and later along the plate separating the new cells, Intracellular localization of PAA-binding sites. probably at the membranes of vesicles and vacuoles, significantly increases, in the smaller daughter cell, destined to later form a polar outgrowth. the first chloronema cell. Thus a system was established to visualize early events in P. patens protoplast polarization at the subcellular level.