Confocal microscopy of double immunofluorescent staining for ChAT and calretinin (CR) or aquaporin 4 (AQP4) in developing somatosensory cortex. A
, B: at P12, the cortical sections immunolabelled for ChAT (red) and CR (blue) show that, after acquisition of serial optical sections, most cholinergic neurons (A
) display also CR immunoreactivity (A
). This is shown in the cells indicated by the white arrows both in A
and in the combination of the two signals (merge, A
). B shows a high magnification detail of ChAT+/CR+ cortical neurons (white arrows). C: at P16, double immunolabelling for ChAT (red) and AQP4 (green) points out a ChAT+ cell (white arrowhead) lying on the wall of one of the numerous vessels (v) identified by the gliovascular marker AQP4. Note also the presence of several cholinergic fibers (red) in the cortical neuropile. Scale bar: 54 μm in A
; 22 μm in B; 17 μm in C.