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. 2022 Mar 3;42(9):1650–1665. doi: 10.1177/0271678X221080324

Figure 2.

Figure 2.

HCAR1 positive cells in dentate gyrus, CA3 and cerebellum. Analysis of brain tissue from reporter mouse line expressing mRFP under the HCAR1 promoter revealed scattered mRFP-positive cells in several regions of the brain. (a) Using a 2-photon microscope, endogenous mRFP signal on fresh 300 µm acute slice was found in the GC layer and the hilus of the DG, in the CA3, and in the Purkinje cell layer of the cerebellum. (b) Using anti-mRFP immunohistochemistry to reveal HCAR1-mRFP positive cells, mRFP expression was found in the hilus of the DG as well as in the inner molecular layer, in the CA3, and in the Purkinje, molecular, and granular cell layer of the cerebellum. Alexa 680 secondary antibody staining is shown in red and nuclei (Hoechst) in blue. (c) Using in situ hybridization (RNAscope™), HCAR1 mRNA transcript was found in the hilus and in the GC layer of the DG, in the CA3, and mainly in the granule and Purkinje cell layer. HCAR1 transcript is shown in red and nuclei (DAPI) in blue. GC = granule cell layer, H = hilus, ML = molecular layer, PC = Purkinje cell.