Fig 2: ClC5 is expressed in amacrine cells, Müller glia, and nNOS-positive cells. A–C, Single z-plane images of chicken retinal tissue sections double-labeled with ClC5 and HPC-1 antibodies reveal that ClC5 is expressed in the HPC-1-positive cells. D–F, Zoom of ClC5 antibody labeling of blue box region in A–C shows that ClC5 is expressed cytosolically in amacrine cells (asterisks). G–I, Zoom of purple box region in A–C shows ClC5 antibody labeling in the ganglion cell layer, indicating that ClC5 is expressed in displaced amacrine cells in the retina (asterisk) as well as in neighboring, HPC-1 negative ganglion cells. J–L, Sections of retinal tissue double-labeled with ClC5 and GS antibodies show that GS and ClC5 co localize in Müller cells (arrows) and in the OLM (arrowheads). M–O, Retinal sections double-labeled with ClC5 and nNOS antibodies reveal expression of ClC5 in nNOS-expressing cells. Yellow arrowheads and asterisks indicate overlapping signal in Müller cells and amacrine cells, respectively. Arrows indicate co-expression in puncta near the INL/IPL border, and white arrowheads indicate co-localization in three bands in the IPL. (Scale A–C, 50 µm, applies to J–L and M–O as well; Scale D–F, 15 µm; Scale G–I, 15 µm.)

Fig 3: Analysis of Cxcl14 co-expression patterns confirms predicted properties Cck.Cxcl14 cells.(A) Cxcl14-expressing cells are CGE derived: in situ hybridization for Cxcl14 combined with immunohistochemistry for YFP in the Lhx6-Cre/R26R-YFP mouse yields no double labeling. (B) Double in situ hybridization for Cxcl14 and Reln marks a population of neurons located primarily at the sr/slm border. Note Reln expression without Cxcl14 in so and slm, likely reflecting O-LM and neurogliaform cells. (C–E) Subsets of the Cxcl14-positive neurons are positive for pro-CCK or CALB1 (in situ hybridization plus immunohistochemistry), or Npy (double in situ hybridization). (F, G) No overlap was seen of Cxcl14 with Nos1 or Kit. In all panels, arrowheads indicate double-expressing neurons. Scale bars: 200 μm (A), 100 μm (B–G). b, sr/slm border region; O-LM, oriens/lacunosum-moleculare; slm, stratum lacunosum-moleculare; sm, stratum moleculare of the dentate gyrus; so, stratum oriens; sp, stratum pyramidale; sr, stratum radiatum; sr/slm, stratum radiatum and stratum lacunosum-moleculare; YFP, yellow fluorescent protein.

Fig 4: Cell body expression of ChR2-mCherry was mostly colocalised with either nNOS or TH in the SNc. Representative images of triple immunolabelling for ChR2-mCherry, nNOS, TH in the SNc, showing exemplar cells exhibiting either: A, colocalisation of nNOS and ChR2-mCherry, but not TH. B, expression of ChR2-mCherry, but neither nNOS nor TH. or C, co-localisation of ChR2-mCherry and TH, but not nNOS. D, Graph shows the mean (+SEM) and individual data points for percentage of ChR2-mCherry that were colocalised with nNOS and/or TH (129 ChR2-mCherry+ cells, five mice). Most ChR2-mCherry+ neurons were either nNOS+ or TH+. Yellow arrows indicate exemplar neurons.

Fig 5: Representative images of ZnT3+ neurons located in OSP of porcine jejunum. All images are composites of merged images taken separately from the blue, red, and green fluorescent channels. a ZnT3 (red) neuron labeled for NOS (green) but SOM (blue)-immunonegative (two small single arrows), and ZnT3 (red) neuron labeled for SOM (blue) but NOS (green)-immunonegative (two single arrows). b ZnT3 (red) neuron labeled for VAChT (green) and VIP (blue)-immunopositive (two small double arrows). c ZnT3 (red) neuron labeled for SP (green) and VIP (blue)-immunonegative (small arrow). Bar 25 µm