Panels AвЂ“O: coronal and sagittal sections taken from the indicated Bregma or medio-lateral coordinates. Double arrows indicate examples of colocalization of two probe mRNAs and single arrows indicate no-co-expression in given cells. Slc17a7, Slc17a6, and Slc32a1: mRNAs encoding the VGLUT1, VGLUT2, and VGAT with respective color coding for the chromogens labeling the corresponding mRNAs. Note: this figure contains excerpts from the more comprehensive figure supplement.ДЄdcyap1, PACAP-mRNA mapping within glutamatergic and GABAergic subpopulations of mouse brain. Nissl staining was used for counterstaining. In these two cerebellar regions, some granule cells also expressed Adcyap1 (indicated with single blue arrows).

Some GABAergic cells in granule cell layer of paraflocculus and central regions also co-expressed Adcyap1 and Slc32a1 (indicated with double pink arrowheads, see also SI-Fig.O1). Purkinje cells are the main GABAergic (expressing Slc32a1, RNA encoding VGAT) PACAP containing neurons, distributed in all regions of cerebellar cortex. Panels E-H show two cerebellar regions, paraflocculus ( E and F) and central ( G and H) lobules, under low and high magnification, respectively, where the Adcyap1 expression was higher than rest of regions.

Double arrows indicate cells co-expressing both mRNAs. Adcyap1 was co-expressed intensely with Slc17a7 (RNA coding for VGLUT1). ( D) Brain stem Koelliker Fuse (KF) nucleus of the parabrachial complex is another main PACAP-expressing nucleus. This feature is better appreciated on the cells in which Adcyap1 is weakly expressed (weak staining), so the Slc17a6 staining can be clearly seen as independent dots. ( B) High magnification of the MEPO region of ( A).В Double arrows indicate cells that co-express both mRNAs. ( A and C) Adcyap1 (RNA coding for PACAP) co-expression with Slc17a6 (RNA coding for VGLUT2) in MEPO and BAC respectively.

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