rnascope multiplex fluorescent v2 detection kit, 323110  (Advanced Cell Diagnostics Inc)

Journal: bioRxiv

Article Title: GPR17 modulates oligodendrocyte precursor cell maturation during development but has limited impact on myelin regeneration following demyelinating insults

doi: 10.1101/2025.07.08.659146

Figure Lengend Snippet: Absence of GPR17 results in a minor enhancement of myelin production during development in vivo and in vitro OPC maturation. Postnatal day 3 mouse brain tissue was used to characterize the genetic GPR17 knockout (Hom) at the mRNA level by in situ hybridization (ISH) and at the protein levels by immunohistochemistry (IHC), in comparison to heterozygous (Het) and wild type (Wt) littermates. GPR17 mRNA and protein were widely expressed throughout the brain in Wt mice, reduced in Het and completely absent in Hom brains (A, top and middle panels). GPR17 was observed in branched cells with OPC morphology, localized in cell processes and neurites as determined by IHC (A, middle panel) and RNAscope (A, bottom, right panel). A few and sparse, thinly myelinated axons were detected in all conditions by means of electron microscopy (A, bottom panels). Studies of cultured OPCs isolated from these mice showed a minor, non-statistically significant increase (unpaired Student’s t-test) in MBP mRNA expression as well as MBP protein expression in cells obtained from GPR17 knockout (Hom) as compared to Het and Wt littermates (B). RQ: relative expression; IHC: immunocytochemistry; ISH: in situ hybridization; Wt: wild type; Het: Heterozygous; Hom: Homozygous; DIV: day in vitro . ISH and IHC bars: 50μm; EM bar: 2μm.

Article Snippet: Final signal detection was carried out using mRNA HRP (RNAscope VS Universal HRP Detection Reagents, ACDBIO) and bluing solution (Roche).

Techniques: In Vivo, In Vitro, Knock-Out, In Situ Hybridization, Immunohistochemistry, Comparison, RNAscope, Electron Microscopy, Cell Culture, Isolation, Expressing, Immunocytochemistry

Journal: bioRxiv

Article Title: GPR17 modulates oligodendrocyte precursor cell maturation during development but has limited impact on myelin regeneration following demyelinating insults

doi: 10.1101/2025.07.08.659146

Figure Lengend Snippet: GPR17 knockout OPCs present minor changes in myelin gene transcripts. Total RNA was isolated from cultured OPCs generated from wild type (Wt) or GPR17 knockout (GPR17 -/- , Het and Hom) newborn pups at the day of plating (D0), and after 1 (D1) and 3 (D3) days in vitro . Whole genome mRNA sequencing was performed, and transcript expression analysis carried out. At all timepoints studied, differentially expressed genes (DEGs) were identified between Wt and GPR17 -/- (A, arrows indicate number of genes being upregulated and downregulated at each time point). GPR17 knockout was confirmed by the complete absence of transcript in the Hom cells (B, top panel). In Wt OPCs, GPR17 is highly expressed at early time points, decreasing significantly at D3. There were no major differences in myelin gene expression (MBP and PLP are shown) between Wt and GPR17 -/- (Hom, B middle and bottom panels). Data are presented as raw p values (Log10) in the volcano plots and as fragments for kilobase of transcript per million (FPKM) in the histograms.

Article Snippet: Final signal detection was carried out using mRNA HRP (RNAscope VS Universal HRP Detection Reagents, ACDBIO) and bluing solution (Roche).

Techniques: Knock-Out, Isolation, Cell Culture, Generated, In Vitro, Sequencing, Expressing, Gene Expression