Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer
|
Author:
Date:
2021-01-05
[Abstract] The subretinal layer between retinal pigment epithelium (RPE) and photoreceptors is a region involved in inflammation and angiogenesis during the procession of diseases such as age-related macular degeneration. The current protocols of whole mounts (retina and RPE) are unable to address the intact view of the subretinal layer because the separation between retina and RPE is required, and each separate tissue is then stained. Non-separate Sclerochoroid/RPE/Retina whole mount staining was recently developed and reported. The method can be further combined and optimized with melanin bleaching and tissue clearing. Here, we describe steps of both non-pigmented and pigmented mouse Sclerochoroid/RPE/Retina whole mount including eyeball preparation, staining, mounting and confocal scanning. In ...
[摘要] [摘要]视网膜色素上皮(RPE)和感光细胞之间的视网膜下层是与年龄相关的黄斑变性等疾病进程中涉及炎症和血管生成的区域。由于需要在视网膜和RPE之间分离,因此整个安装(视网膜和RPE)的当前方案无法解决视网膜下层的完整视图,然后对每个单独的组织进行染色。最近开发并报道了非分离的巩膜脉络膜/ RPE /视网膜整装染色。该方法可以与黑色素漂白和组织清除进一步结合和优化。在这里,我们描述了非色素和色素小鼠硬化脉络膜的步骤/ RPE /视网膜整个安装,包括眼球准备,染色,安装和共聚焦扫描。此外,我们提出了巩膜脉络膜/ RPE /视网膜整个支架中的RPE,视网膜下小胶质细胞和邻近的感光体的共聚焦图像。
[背景]在眼睛视网膜是由视网膜色素上皮(RPE)包围,脉络膜和巩膜。通常,在整装染色中,视网膜组织与脉络膜/ RPE分开,并且分开的视网膜和脉络膜/ RPE的每个部分都被染色。因此,视网膜或脉络膜/ RPE的单独的整体染色不能解决完整的视网膜下信息。最近,开发了脉络膜/ RP E /视网膜整个安装方案(Kim等,2016; ...
|
|
In vivo and in vitro 31P-NMR Study of the Phosphate Transport and Polyphosphate Metabolism in Hebeloma cylindrosporum in Response to Plant Roots Signals
|
Author:
Date:
2018-08-20
[Abstract] We used in vivo and in vitro phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy to follow the change in transport, compartmentation and metabolism of phosphate in the ectomycorrhizal fungus Hebeloma cylindrosporum in response to root signals originating from host (Pinus pinaster) or non-host (Zea mays) plants. A device was developed for the in vivo studies allowing the circulation of a continuously oxygenated mineral solution in an NMR tube containing the mycelia. The in vitro studies were performed on fungal material after several consecutive treatment steps (freezing in liquid nitrogen; crushing with perchloric acid; elimination of perchloric acid; freeze-drying; dissolution in an appropriate liquid ...
[摘要] 我们使用体内和体外磷-31核磁共振( 31 P-NMR)光谱来跟踪运输,分区和 外生菌根真菌 Hebeloma cylindrosporum 中的磷酸盐代谢响应来自宿主( Pinus pinaster )或非宿主( Zea mays )的根信号植物。 开发了一种用于体内研究的装置,其允许连续氧化的矿物质溶液在含有菌丝体的NMR管中循环。 在几个连续的处理步骤(在液氮中冷冻;用高氯酸压碎;消除高氯酸;冷冻干燥;在适当的液体培养基中溶解)后,对真菌材料进行体外研究。
【背景】 菌根真菌和植物之间的关联改善了宿主植物的P营养(Smith和Read,2008; Plassard和Dell,2010; Cairney,2011; Smith 等人,,2015)。这种积极效应主要归因于真菌菌丝对磷酸盐(Pi)的吸收,探测了在活跃吸收根周围的耗竭区以外的大量土壤(Smith和Read,2008; Cairney,2011; Smith et al。< em="">,2015)和真菌细胞分泌细胞外磷酸酶(Quiquampoix和Mousain,2005)。吸收的Pi部分地掺入磷酸化的代谢物,磷脂和核酸中,并且部分地浓缩成多磷酸盐(PolyP),其中它们构成液泡中的储存池(Ashford 等人,,1994)。该协议详述了一种装置,该装置允许通过 31 ...
|
|
Establishing a Symbiotic Interface between Cultured Ectomycorrhizal Fungi and Plants to Follow Fungal Phosphate Metabolism
|
Author:
Date:
2017-10-20
[Abstract] In ectomycorrhizal plants, the fungal cells colonize the roots of their host plant to create new organs called ectomycorrhizae. In these new organs, the fungal cells colonize the walls of the cortical cells, bathing in the same apoplasm as the plant cells in a space named the ‘Hartig net’, where exchanges between the two partners take place. Finally, the efficiency of ectomycorrhizal fungi to improve the phosphorus nutrition of their host plants will depend on the regulation of phosphate transfer from the fungal cells to plant cells in the Hartig net through as yet unknown mechanisms. In order to investigate these mechanisms, we developed an in vitro experimental device mimicking the common apoplasm of the ectomycorrhizae (the Hartig net) to study the phosphorus metabolism in the ...
[摘要] 在外生菌根植物中,真菌细胞定植于其宿主植物的根部,以产生称为外生菌根的新器官。在这些新器官中,真菌细胞定居在皮质细胞的壁上,与称为“Hartig网”的空间中的植物细胞在同一质粒中沐浴,两地之间的交流发生。最后,外源菌根真菌提高其宿主植物的磷营养的效率将取决于通过尚未知的机制调节从真菌细胞到Hartig网中植物细胞的磷酸转移。为了研究这些机制,我们开发了一种体外实验装置,模拟外生菌根(Hartig网)的常见质粒来研究外生菌根真菌中的磷代谢。 当真菌细胞与寄主植物“Pinus pinaster”的植物细胞相关联时。该装置可用于监测从先前与 32 P-正磷酸盐一起培养的真菌的磷酸盐外排。 【背景】已知菌根真菌和植物之间的关联可以改善植物P营养(由Smith和Read,2008; Plassard和Dell,2010; Cairney,2011; ...
|
|