Journal: The Journal of Neuroscience

Article Title: Kv4.1, a Key Ion Channel For Low Frequency Firing of Dentate Granule Cells, Is Crucial for Pattern Separation

doi: 10.1523/JNEUROSCI.1541-19.2020

Figure Lengend Snippet: Comparison between Kv4.1 and Kv4.2 expression in the hippocampus. A, Both Kv4.1 and Kv4.2 antibodies selectively recognize the channel in HEK293 cells. HEK293 cells were transfected with either AcGFP-tagged Kv4.1 cDNA (Kv4.1-AcGFP Tf) or Kv4.2 (Kv4.2-AcGFP Tf). Forty-eight hours after transfection, the cells were stained using Kv4.1 or Kv4.2 antibodies. Scale bar, 20 μm. B, Representative fluorescence immunostaining images show that Kv4.1 (top) is highly expressed in DG while Kv4.2 (bottom) is broadly expressed in the hippocampus of 8-week-old mice. Scale bars, 500 μm (left) and 50 μm (middle and right). C, Summary bar graphs of the relative fluorescence intensity (F − F0) shown in the DG and CA1 for Kv4.1 (n = 10, left) and Kv4.2 (n = 5, right). (Kv4.1; GCL, 20.3 ± 2.0; ML, 11.9 ± 1.3; Pyr, 12.0 ± 2.3; Rad, 11.9 ± 2.2; GCL vs ML, p = 0.0037; GCL vs Pyr, p = 0.013; Pyr vs Rad, p = 0.97; Kv4.2; GCL, 24.2 ± 2.3; ML, 39.1 ± 5.8; Pyr, 25.1 ± 2.3; Rad, 33.5 ± 3.3; GCL vs ML, p = 0.037; Pyr vs Rad, p = 0.016). D, Western blot analysis for Kv4.1 and Kv4.2 in DG and CA1 of 8-week-old mice. Right, DG exhibits higher Kv4.1 expression level compared with CA1; 1.00 ± 0.02 in DG, 0.53 ± 0.05 in CA1, n = 8; p < 0.0001. Paired t test. Prospero-related homeobox 1 (Prox) and α-tubulin used as a marker for DG and loading control, respectively. *p < 0.05, **p < 0.01, ***p < 0.001, N.S. (not significant) p > 0.05 by Student's t-test.

Article Snippet: The following antibodies were purchased from commercial sources: anti-Kv4.3, APC-017, Alomone; anti-Prox1 antibody, PRB-238C-200, BioLegend; anti-α-tubulin, T5168, Sigma-Aldrich.

Techniques: Comparison, Expressing, Transfection, Staining, Fluorescence, Immunostaining, Western Blot, Marker

Journal: Stem Cells (Dayton, Ohio)

Article Title: P2X7 Receptors Regulate Phagocytosis and Proliferation in Adult Hippocampal and SVZ Neural Progenitor Cells: Implications for Inflammation in Neurogenesis

doi: 10.1002/stem.2894

Figure Lengend Snippet: Adult NPCs were characterized as type 2/type C intermediate progenitor cells and express P2X7 receptors in vitro. Representative NPCs derived from the hippocampus and cultured in vitro were stained for GFAP and nestin (A–C) to identify type 1 NPCs. NPCs were also positive for the markers SOX2, vimentin (D–F), and BLBP ( G–H ). MASH1 was used to identify type 2 NPCs ( G, I ) and had a variable expression from low (indicated by hash) to high (asterisk). NPCs derived from the hippocampus expressed Prox1, a marker for granule cell neurons, and all NPCs expressed intracellular P2X7 receptors ( J–L ). Approximately 10% of representative hippocampal NPCs displayed negative P2X7 immunoreactivity under nonpermeabilized conditions, for example, they did not express P2X7 receptors on the membrane surface, indicated by an asterisk ( M–O ). Dual labeling against intracellular P2X7 and extracellular P2X7 revealed cells with negative surface expression retained intracellular stores ( P–R ). N ≥ 3; scale bars represent 10 μm.

Article Snippet: Primary antibodies were applied for 16 hours at 4°C and included those raised against P2X7 (rabbit, 1:1000, Alomone Labs, Jerusalem, Israel intracellular: APR‐004; and extracellular: APR‐008, epitopes), PROX1 (mouse, 1:200), glial fibrillary acidic protein (GFAP, mouse conjugate, 1:1000), nestin (rabbit, 1:200, Abcam, Cambridge, UK); SRY‐related HMG BOX gene 2 (SOX2, rabbit, 1:2000, Abcam), vimentin (mouse, 1:1000), mammalian achaete‐scute homolog 1 (MASH1, mouse, 1:500, BD Pharmingen, San Jose, CA), brain lipid‐binding protein (BLBP, rabbit, 1:1000), and doublecortin (DCX, rabbit, 1:200, Cell Signaling Technology, Danvers, MA).

Techniques: In Vitro, Derivative Assay, Cell Culture, Staining, Expressing, Marker, Labeling