Article Figures & Data
Figures
- Figure 1.
Immunocytochemical detection of ChC axo-axonic connections. Innervation of AISs by ChC cartridges at different ages. Top, z-projection images of a complete ChC arbor (green, GFP) and AISs (red, AnkyrinG and PIkB immunolabeling) from Nkx2.1-Cre::Madm mice at P18, P30, and P90. Bottom, Projection of 3D reconstructions of the ChC arbors. Green represents cartridges, yellow represents an AIS apposed by a cartridge, and red represents AISs that are not apposed by a cartridge. Scale bar, 50 μm.
- Figure 2.
Morphological properties of ChC arbors. A, Number of boutons per cartridge. The majority of the cartridges were found to contain three to five boutons. B, C, Distribution of cartridges (B) and AISs (C) within the ChC arbors examined using Sholl analysis. The cell body is located at the center (radius = 0 μm), and the percentage of cartridges or AISs is determined at each radial ring around the cell body at 30 μm intervals. No significant differences were found for the percentage distributions of cartridges and AISs at different ages, and data from all ages are combined (n = 18, 6 cells per each age).
- Figure 3.
Dense ChC axo-axonic innervation. A, Apposition of cartridges and AISs. Single z-plane image of a ChC cartridges (green) and AISs (red) at P30. Inset, Area with a high density of cartridges, where almost every AIS is apposed by a cartridge. Scale bar, 50 μm. B, Sholl analysis of percentage of cartridges apposing an AIS within chandelier arbors. Almost every cartridge apposes an AIS. C, Maximum Sholl analysis of the percentage of AISs apposed by a cartridge within a chandelier arbor. The ChC arbor is defined by the maximum x, y, and z coordinates. D, Convex hull Sholl analysis of the percentage of AISs apposed by a cartridge. The ChC arbor is limited to the coordinates where a cartridge is present.
- Figure 4.
Conservation of ChC innervation patterns at P18, P30, and P90. A–E, Convex hull Sholl analysis revealed no significant difference in the distribution of cartridges (A), boutons (B), AIS (C), or appositions (D, E) at different ages.
- Figure 5.
Overlapping innervation of AISs by ChCs. A, Confocal image of the axonal arbor of an individual GFP+ ChC (green) and Ankyrin G-ir AISs at P30. Rows of GFP+ buttons (ChC cartridges) are in apposition to Ankyrin G-ir AIS (arrows). B, VGAT-ir buttons (red) in the same field as in A. C, Combination of A and B (stacks of 34 optical sections; step size: 0.38 μm) to measure the number of ChCs that innervate each AIS. D, Schematic representation of the innervated Ankyrin G-ir AISs (blue lines) illustrated in C with rows of GFP+ buttons (green dots) in apposition to them. Red dots represent VGAT-ir buttons in contact with the AISs (putative GABAergic synapses from other ChCs; see Discussion for further explanation). Scale bar: D (for A–D), 9 μm.
