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Abstract

Cover: In Erben and Buonanno (https://doi.org/10.1002/cpns63), the image shows examples for chromogenic BaseScope detection (Basic Protocol 2) and ISH-IHC combination (Basic Protocol 3). (A) Hippocampal section of an adult WT mice, labeled using BaseScope for ErbB4 with a single probe pair targeting the exon boundary exon 1/exon 2. The section was counterstained with hematoxylin and signal detected with light microscopy. Arrowheads indicate ErbB4+ GABAergic interneurons. (B) Detection of one of the four ErbB4 splice variants (JMb; cyan) by BaseScope in GABAergic interneurons (GAD-GFP; green; arrowheads) in a section of an adult GAD-GFP mouse (kindly provided by Dr. Yuchio Yanagawa). GFP signal was amplified after the ISH assay with an anti-GFP antibody (NeuroMab; N86/8; RRID: AB_10671444). (C) ISH for ErbB4 (C1; cyan) and Th (C3; magenta) was combined with IHC with an antibody against DAT (green; Santa Cruz, sc-32258; RRID: AB_627400) in a sagittal FFPE section from an adultWT mouse; depicted substantia nigra compacta (SNc) on the left and dopaminergic medial forebrain bundle. (C′) Magnification of ErbB4-expressing dopaminergic neurons (arrowheads) in the SNc. (D) In primary mesencephalic cultures (DIV8; prepared as in Skirzewski et al., 2018), DAT (green) immunostaining was performed post-hoc to RNAscope for ErbB4 (C1; cyan) and Th (C3; magenta). Scale bars 100 µm in B, 20 µm in other panels. Abbreviations: ISH-IHC, in situ hybridization-immunohistochemistry; GFP, green fluorescent protein; FFPE, formalin-fixed paraffin-embedded; WT, wild type.