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. Author manuscript; available in PMC: 2022 May 9.

Published in final edited form as: Mol Neurobiol. 2021 Oct 19;59(1):245–265. doi: 10.1007/s12035-021-02511-w

Fig. 3 — Optogenetic stimulation of MCH shortens cilia length. a Experimental approach. Adult PmchCre + and PmchCre− mice were stereotaxically injected with AAV-EF1α-DIO-ChR2-T159c-eYFP in the lateral hypothalamus (LH). A fiber cannula was then placed in the LH slightly above the injection site. Diagram was created with BioRender.com webpage. b Coexpression of ChR2 (green) containing neurons in the lateral hypothalamus and MCH immunofluorescence (red) and counterstained with DAPI (blue) in PmchCre + and PmchCre− mice. (scale bar = 10 μm, magnification = 40×). c ChR2 fluorescence (green) and c-Fos immunofluorescence (red) identifies cells recently activated in PmchCre + and PmchCre− mice (scale bar = 10 μm). d, g, j, m Immunostaining of ADCY3 labeled in red fluorescence in the d PFC, g CA1, j CPu, and m NAc after blue light activation in PmchCre + and PmchCre− mice (scale bar = 10 μm). e, h, k, n Quantification of the length of ADCY3 + primary cilia (μm) in the e PFC, h CA1, k CPu, and n NAc, *P < 0.05, **P < 0.01, and ***P < 0.001. f, I, l, o Cilia were grouped by length (μm) and plotted against the number of cells (%) in the f PFC, i CA1, l CPu, and o NAc

Fig. 3 — Optogenetic stimulation of MCH shortens cilia length. a Experimental approach. Adult PmchCre + and PmchCre− mice were stereotaxically injected with AAV-EF1α-DIO-ChR2-T159c-eYFP in the lateral hypothalamus (LH). A fiber cannula was then placed in the LH slightly above the injection site. Diagram was created with BioRender.com webpage. b Coexpression of ChR2 (green) containing neurons in the lateral hypothalamus and MCH immunofluorescence (red) and counterstained with DAPI (blue) in PmchCre + and PmchCre− mice. (scale bar = 10 μm, magnification = 40×). c ChR2 fluorescence (green) and c-Fos immunofluorescence (red) identifies cells recently activated in PmchCre + and PmchCre− mice (scale bar = 10 μm). d, g, j, m Immunostaining of ADCY3 labeled in red fluorescence in the d PFC, g CA1, j CPu, and m NAc after blue light activation in PmchCre + and PmchCre− mice (scale bar = 10 μm). e, h, k, n Quantification of the length of ADCY3 + primary cilia (μm) in the e PFC, h CA1, k CPu, and n NAc, *P < 0.05, **P < 0.01, and ***P < 0.001. f, I, l, o Cilia were grouped by length (μm) and plotted against the number of cells (%) in the f PFC, i CA1, l CPu, and o NAc