Gimenez-Molina Yolanda, Villanueva José, Nanclares Carmen, Lopez-Font Inmaculada, Viniegra Salvador, Francés Maria Del Mar, Gandia Luis, Gil Amparo, Gutiérrez Luis M
Instituto de Neurociencias, Centro Mixto del Consejo Superior de Investigaciones Científicas, Universidad Miguel HernándezAlicante, Spain.
Instituto Teófilo Hernando, Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de MadridMadrid, Spain.
Front Cell Neurosci. 2017 May 4;11:135. doi: 10.3389/fncel.2017.00135. eCollection 2017.
Cultured bovine chromaffin cells have been used extensively as a neuroendocrine model to study regulated secretion. In order to extend such experimental findings to the physiological situation, it is necessary to study mayor cellular structures affecting secretion in cultured cells with their counterparts present in the adrenomedullary tissue. F-actin concentrates in a peripheral ring in cultured cells, as witnessed by phalloidin-rodhamine labeling, while extends throughout the cytoplasm in native cells. This result is also confirmed when studying the localization of α-fodrin, a F-actin-associated protein. Furthermore, as a consequence of this redistribution of F-actin, we observed that chromaffin granules and mitochondria located into two different cortical and internal populations in cultured cells, whereas they are homogeneously distributed throughout the cytoplasm in the adrenomedullary tissue. Nevertheless, secretion from isolated cells and adrenal gland pieces is remarkably similar when measured by amperometry. Finally, we generate mathematical models to consider how the distribution of organelles affects the secretory kinetics of intact and cultured cells. Our results imply that we have to consider F-actin structural changes to interpret functional data obtained in cultured neuroendocrine cells.
培养的牛嗜铬细胞已被广泛用作神经内分泌模型来研究调节性分泌。为了将这些实验结果扩展到生理情况,有必要研究培养细胞中影响分泌的主要细胞结构及其在肾上腺髓质组织中的对应结构。通过鬼笔环肽-罗丹明标记可以看到,F-肌动蛋白在培养细胞中集中在周边环中,而在天然细胞中则延伸至整个细胞质。在研究与F-肌动蛋白相关的蛋白α-血影蛋白的定位时,这一结果也得到了证实。此外,由于F-肌动蛋白的这种重新分布,我们观察到嗜铬颗粒和线粒体在培养细胞中位于两个不同的皮质和内部群体中,而在肾上腺髓质组织中它们均匀地分布在整个细胞质中。然而,通过安培法测量时,分离细胞和肾上腺组织块的分泌非常相似。最后,我们生成数学模型来考虑细胞器的分布如何影响完整细胞和培养细胞的分泌动力学。我们的结果表明,我们必须考虑F-肌动蛋白的结构变化来解释在培养的神经内分泌细胞中获得的功能数据。