Journal: The Journal of Neuroscience

Article Title: Interdependent Conductances Drive Infraslow Intrinsic Rhythmogenesis in a Subset of Accessory Olfactory Bulb Projection Neurons

doi: 10.1523/JNEUROSCI.2520-15.2016

Figure Lengend Snippet: Expression of BK and CaV2.3 channels in AOB mitral cells. A–C, Confocal laser-scanning microscopy images of the mouse AOB (parasagittal cryosections). Immunofluorescence labeling (green) of CaV2.3 (A), the BK channel subunit α-1 (Sloα1; B), and the BK channel subunit β-4 (Sloβ4; C). Nuclei are counterstained with DRAQ5 (red). Dashed lines indicate both the lateral olfactory tract (LOT) and the glomerular layer (GL) in Aiii, Biii, and Ciii. Representative single optical section images show an overview of the AOB (Ai, Bi, Ci) and a high-magnification view (Aii, Bii, Cii) of the mitral cell layer (MCL). Note the robust labeling of cells in the MCL as well as the absence of immunofluorescence following peptide preadsorption (Aiii, Biii, Ciii; no DRAQ5 counterstaining). D, E, Post hoc immunolabeling of previously recorded intrinsically rhythmic neurons. Confocal images of AOB cryosections containing biocytin-filled iAMC somata and proximal dendrites (visualized by Alexa Fluor 488/633 streptavidin conjugate). Sections were counterstained against the BK channel α-1 (D) and β-4 (E) subunits, respectively. Merged images (Diii, Eiii) reveal BK expression in these representative iAMCs. Div, Eiv, Western blot analysis of total olfactory bulb (right lanes) and posterior brain (left lanes) protein extracts. Immunoblots were probed with anti-Sloα1 (D) and anti-Sloβ4 (E) antibodies, respectively. Expected band size is indicated above both blots. d, Dorsal; FC, frontal cortex; v, ventral.

Article Snippet: Primary antibodies were anti-Slo1 (K Ca 1.1, 1:500; Alomone Labs), anti-Sloβ4 (1:100; Alomone Labs), and anti-Ca V 2.3 (1:100; Alomone Labs).

Techniques: Expressing, Confocal Laser Scanning Microscopy, Immunofluorescence, Labeling, Immunolabeling, Western Blot

Journal: The Journal of Neuroscience

Article Title: Somatic Localization of a Specific Large-Conductance Calcium-Activated Potassium Channel Subtype Controls Compartmentalized Ethanol Sensitivity in the Nucleus Accumbens

doi: 10.1523/JNEUROSCI.0684-04.2004

Figure Lengend Snippet: NAcc neurons express BK channel α subunits. A, Hoechst staining shows cell nuclei of BK channel α subunit in stably transfected HEK293 cells. B, The same neurons as in A are stained with the BK channel α subunit, using a specific polyclonal antibody, visualized by fluorescence microscopy (60×). C, Hoechst staining shows cell nuclei of untransfected HEK293 cells. D, BK channel α subunit antibody fails to stain untransfected HEK cells shown in C. E, Staining of α subunit in the core NAcc (10×; the anterior commissure is not shown). In the bottom left corner, the white box outlines a region shown at higher magnification (40×) in F. F, Single and double arrowheads point to labeled primary and secondary processes, respectively, indicating that α subunits are not confined to the soma but are also expressed in neuronal processes.

Article Snippet: Sections were incubated (overnight at 4°C) with specific polyclonal primary antibodies (1:300) against BK channel subunits [α, β1 (Affinity BioReagents, Golden, CO) and β4 (gifts from Millennium Pharmaceuticals, Cambridge, MA)], rinsed, and incubated for 1 hr at room temperature with Alexa 594-tagged, anti-rabbit secondary antibody (1:400; Molecular Probes, Eugene, OR).

Techniques: Staining, Stable Transfection, Transfection, Fluorescence, Microscopy, Labeling