Organization of the core respiratory network: Insights from optogenetic and modeling studies
Fig 2
Glycine and GABA antibody, and ChR2-EYFP expression in Cre-dependent tdTomato-labeled neurons in the pre-BötC region in VGAT-tdTomato or VGAT-tdTomato-ChR2-EYFP mouse strains.
(A) Confocal fluorescence microscopy images of the pre-BötC region at low magnification (upper panels) and a pre-BötC subregion at higher magnification (lower panels) in a representative fixed coronal section from an adult VGAT-tdTomato mouse showing VGAT-Cre dependent tdTomato-labeled neurons (red) and glycine antibody labeling of neurons (green), and co-localization (arrows) of glycine antibody labeling in VGAT-expressing pre-BötC neurons in merged images (Yellow). Example of VGAT-expressing tdTomato-labeled neuron without glycine antibody expression (arrowhead) in this histological section is also indicated. (B) Confocal fluorescence microscopy images of a subregion within the pre-BötC in a fixed coronal section from the VGAT-tdTomato mouse line, showing VGAT-Cre dependent tdTomato labeling (red) and GABA antibody labeling (green) in pre-BötC neurons. Merged image illustrates co-localization of GABA antibody labeling in tdTomato-labeled neurons. Examples of tdTomato-labeled neurons without GABA antibody expression (arrowheads) are also indicated. (C) Two-photon microscopy single optical plane “live” images of the pre-BötC subregion in an in vitro neonatal medullary slice from the VGAT-tdTomato-ChR2-EYFP mouse line, illustrating that tdTomato-labeled VGAT-positive pre-BötC neurons (red) express ChR2-EYFP (green) in somal membranes, as confirmed in the merged image. All images have the same dorso-medial anatomical orientation. Abbreviations: d, dorsal; m, medial; NA, nucleus ambiguus; VS, ventral surface.