Slice Cultures as a Model to Study Neurovascular Coupling and Blood Brain Barrier In Vitro
Figure 2
Ca2+-imaging in perivascular and parenchymal astrocytes. (a) 3D reconstruction of the astrocytic network covering a vessel after colabeling the slice culture with calcein-AM (green fluorescence channel) and rhod-2 AM (red fluorescence channel). Note the considerable cytosolic rhod-2 fluorescence besides the presence of the rhod-2 labeled mitochondria in the neuropil. The excerpt on the left shows the cross-section of the same vessel. (b) Z-series of confocal images (1.2 μm steps) from the same vessel were used to distinguish between perivascular and parenchymal astrocytes, that is, with or without contact to the wall of the vessel. Scale bars in (a) and (b) represent 10 μm. (c) Comparison of [Ca2+]i transients between perivascular and parenchymal astrocytes during low Mg2+-ACSF induced epileptiform activity. Seizure-like events (lower trace: field potential) were associated with slight elevation of astrocytic [Ca2+]i and were followed by large amplitude [Ca2+]i transients. There were no statistical differences in amplitude or duration of [Ca2+]i transients between perivascular and parenchymal astrocytes.
