Journal: Frontiers in molecular neuroscience

Article Title: Specific Role for GSK3α in Limiting Long-Term Potentiation in CA1 Pyramidal Neurons of Adult Mouse Hippocampus.

doi: 10.3389/fnmol.2022.852171

Figure Lengend Snippet: FIGURE 1 | Spatial validation of GSK3α or GSK3β cKO by IHF. (A) Fluorescence images of brain sections through the hippocampus collected with a 10 × objective lens following staining with anti-GSK3α (revealed with Alexa 594) and anti-GSK3β (revealed with Cy5), as well as DNA using DAPI; the three columns on each row is from a single section imaged sequentially in the corresponding fluorescence wavelength channels. IHF from each of a GSK3α cKO, GSK3β cKO, and a Cre- subject are shown in rows. Note the prominent reduction in corresponding GSK3 staining within the CA1 region of each paralog cKO. (B) The white outlined zones in A were subsequently imaged with a 60 × lens and confocal optical resolution. The same fluorescence channels are presented, plus a final merged overlay in the final column. (C) Primary antibody was omitted, and images from the same Cre- subject were collected at the exact same settings as those used in (A). (A–C) Images shown were selected from a sampling of n = 2–3 subjects per genotype, and are representative from a total of 36 images collected with a 10 × objective lens, and 82 images collected with a 60 × objective lens.

Article Snippet: Slides were washed with PBS 3 × 5 min, incubated with secondary antibodies, donkey anti-rabbit conjugated with Alexa Fluor 594 (Jackson ImmunoResearch Inc, West Grove, PA, United States, Cat# 711- 585-152) and donkey anti-mouse conjugated with Cy5 (Jackson ImmunoResearch Inc, Cat# 715-175-150) at 1:1000 diluted with PBS for 1 h at room temperature.

Techniques: Biomarker Discovery, Fluorescence, Staining, Sampling