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. 2010 Dec 23;519(6):1007–1023. doi: 10.1002/cne.22570

Figure 9.

Figure 9

NPY axons that innervate NK1r lamina III cells and PKCγ lamina II interneurons appear to originate from different sources. a,b: Part of laminae II and III in a sagittal section that had been immunostained for NPY (red), NK1r (green), and PKCγ (blue). Several PKCγ-immunoreactive neurons can be seen and five of these are numbered. Each of these cells has contacts from NPY-immunoreactive boutons. Some of these are marked with arrows and shown at higher magnification in c–f. A large NK1r-immunoreactive cell is also present in this field and this has numerous contacts from NPY boutons. c–f: Single confocal optical sections scanned to reveal NPY (red), PKCγ (blue), and gephyrin (white) show that a gephyrin punctum is present at each of the contacts that the PKCγ cells receive from NPY boutons (arrows). g,h: Show contacts from NPY-containing boutons onto one of the dorsal dendrites of the NK1r cell at higher magnification and correspond to the boxed region in b. Seven of the NPY boutons that contact the cell are identified with numbers. The images in the bottom row, each of which is from a single optical section, are in pairs. In each case the left one was scanned for NPY (red), NK1r (green), and gephyrin (white), while the right one shows only gephyrin. Each of the contacts between the NPY boutons and the dendrite of the NK1r cell is associated with a gephyrin punctum, but these are much paler than those on the PKCγ cells. Note in h that the NPY-immunoreactive boutons that contact the NK1r cell are often linked by clear intervaricose portions, which indicates that they must originate from the same neuron. However, these axons also give rise to boutons that are not in contact with the NK1r cell (an example is indicated with an arrow in g,h). The NPY boutons that are presynaptic to the PKCγ cells are generally paler (as shown in a), and were never found to be connected by visible intervaricose portions to those that were presynaptic to the NK1r cells. a,b: Projections of 34 optical sections and g,h: of 8 optical sections at 0.5 μm z-spacing. Scale bars = 20 μm in a,b; 10 μm in c–h.