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. 2004 Jul 29;560(Pt 1):231–247. doi: 10.1113/jphysiol.2004.070060

Figure 11. Spatial (3-D) distribution of presynaptic VIP-LI and VAChT-LI appositions (A and B, respectively) on IMG neurones, and neuronal MAb35 immunoreactivity (putative nAChRs) on the surface (C) and inside the same neurone (D).

Figure 11

IMG neurones were intracellularly injected with LY, immunostained with appropriate antibodies, visualized with either Cy3 or Cy5, and imaged using a confocal microscope. The images of the LY-filled neurones (coloured grey) and associated immunoreactive sites (coloured red) were volume reconstructed and superimposed using ANALYZE™ software (see Methods for details). Note that the majority of VIP-LI presynaptic endings (55.9 ± 5.7% of the total neurone-associated VIP LI-containing structures) were on secondary and tertiary dendritic branches of the neurone. Note that a majority of VAChT-LI (marker of the presynaptic ACh-containing terminals) and majority of MAb35 sites (marker of the nAChRs) were also found on secondary and tertiary dendrites (see Table 2 for quantification). The presence of MAb35-LI sites within the IMG neurone (panel D) most likely represents nAChR subunits associated with intracellular organelles during biosynthesis and trafficking of nAChRs (Ermilov et al. 2003). Scale bars, 50 μm.