Uckermann Ortrud, Iandiev Ianors, Francke Mike, Franze Kristian, Grosche Jens, Wolf Sebastian, Kohen Leon, Wiedemann Peter, Reichenbach Andreas, Bringmann Andreas
Paul Flechsig Institute of Brain Research, Department of Neurophysiology, University of Leipzig, Leipzig, Germany.
Glia. 2004 Jan 1;45(1):59-66. doi: 10.1002/glia.10305.
Müller glial cells within the retina may respond to different signaling molecules with an elevation of their intracellular free calcium. To prove the localization of the recorded calcium responses in Müller cells within acutely isolated retinal wholemounts, retinal pieces from adult animals and humans were exposed to different vital dyes just after the calcium imaging records were finished. The dyes, Mitotracker Orange, Mitotracker Green, Celltracker Orange, Celltracker Green, and monochlorobimane, are all selectively taken up by Müller glial cells, while neuronal cells remain largely devoid of the dyes. By using this method, it can be demonstrated that the free calcium alterations within the wholemounts indeed occur within Müller cells. Moreover, the cross-sectional areas of (dye-filled) Müller glial cell bodies, as well as of (dye-free) neuronal cell bodies, can be measured in retinal wholemounts, and the spatial densities of both types of cells can be determined. The vital dye loading of Müller cells may facilitate investigations of stimulus-induced alterations of retinal glial cell physiology and morphology.
视网膜内的米勒胶质细胞可能会对不同的信号分子作出反应，使其细胞内游离钙升高。为了证实急性分离的视网膜全层标本中所记录的钙反应在米勒细胞中的定位，在钙成像记录完成后，将成年动物和人类的视网膜切片暴露于不同的活性染料中。这些染料，如线粒体追踪橙、线粒体追踪绿、细胞追踪橙、细胞追踪绿和单氯联苯二胺，都能被米勒胶质细胞选择性摄取，而神经元细胞基本不会摄取这些染料。通过使用这种方法，可以证明全层标本中的游离钙变化确实发生在米勒细胞内。此外，在视网膜全层标本中，可以测量（充满染料的）米勒胶质细胞体以及（无染料的）神经元细胞体的横截面积，并确定这两种细胞的空间密度。米勒细胞的活性染料加载可能有助于研究刺激诱导的视网膜胶质细胞生理和形态变化。
