G,N: a medium spiny neuron in the striatum. (arrows). Scale bar on panel B for panels A-D: 10 m; scale bar on panel F for panels E-H: 5 m (2X magnified). All images were obtained using Apotome structured illumination microscopy. A red-green version of this figure is available in the main body of the paper as Figure 1.Figure 2. Kv2.1 is localized at AnkG-deficient sites on the AIS of rat layer 5 neocortical pyramidal neurons. Rat brain sections double immunofluorescence labeled for Kv2.1 (green) and AnkG (magenta). A-F: Images obtained with a Zeiss Elyra super resolution microscope, showing two examples (A-C and D-F) of double labeling (A,D), and the AnkG (B,E) and Kv2.1 (C,F) signals alone. Arrows in panels correspond to same locations on each panel. G-R: Images showing double labeling (G,I,K), and the AnkG signal alone (H,J,L). G,H: Images obtained with a Zeiss Apotome microscope. I,J: Images obtained with an Olympus ML241 confocal microscope. K,L: Images obtained with a Nikon N-SIM microscope. Panels below M-R are 4X-magnified images of the area demonstrated in the boxes in panels G-L above. Graphs below panels M-R are histograms of fluorescence intensity across the collection drawn on each panel. Scale pub on panel D for panels A-F: 2 m. Level bar on panel H for panels G-L: 5 m. Level bar on panel H for panels G,H, on panel J for panels I,J, on panel L for panels K,L; on panel N for panels M,N, on panel P for panels O,P, and on panel R for panels Q,R: 1.25 m. A red-green version of this number is available in the main body of the paper as Number 2. Number 3. Kv2.1 is localized at AnkG-deficient sites within the AIS of neurons in different regions of rat mind. Rat mind sections double immunofluorescence labeled for Kv2.1 (green) and AnkG (magenta). Images were from neurons in different mind areas. A-D, H-K: hippocampus. A,H: CA1 pyramidal neurons; B,I: a parvalbumin-negative interneuron in of CA1; C,J: a parvalbumin-positive interneuron in of CA1; D,K: dentate granule cells. E,F,L,M: thalamus. E,L: a neuron in the posterior nucleus; F,M: a neuron in the lateral posterior nucleus. G,N: a medium spiny neuron in the striatum. Arrows in panels correspond to the location of the midpoint of the 4X enlarged ML241 insets in panels H-N. Graphs below panels H-N are histograms of fluorescence intensity across the collection drawn on each panel. Scale pub on panel A for panels A-G: 5 m; Level bar on panel H for panels H-N: 1 m (4X ML241 magnified). All images were acquired using Apotome organized illumination microscopy. A Mouse monoclonal to HDAC4 red-green version of this number is available in the main body of the paper as Number 3. Number 4. Kv2.1 is localized at AnkG-deficient sites within the AIS of coating 5 neocortical pyramidal neurons in different mammalian varieties. Sections double immunofluorescence labeled for Kv2.1 (green) and AnkG (magenta). Images were from neocortical neurons in the brains of different mammalian varieties: A,F: rat; B,G: ferret; C,H: monkey; D,E,I,J: human being. Arrows in panels A-E correspond to the location of the midpoint of the 4X enlarged insets demonstrated as panels F-J, respectively. Graphs below panels F-J are histograms of fluorescence intensity across the collection drawn on each panel. Scale pub on panel E for panels A-E: 5 m; Level bar on panel J for panels F-J: 1 m (4X magnified). All images were acquired using Apotome organized illumination microscopy. A red-green version of this number is available in the main body of the paper as Number 4. NIHMS560816-supplement-Supp_Numbers1-S4.pdf (938K) GUID:?33592216-81A3-454D-8DDB-5E7B15134129 Abstract The axon initial segment (AIS) plays a key role in initiation of action potentials and neuronal output. The plasma membrane of the AIS consists of high densities of voltage-gated ion channels required for these electrical events, and much recent work offers focused on defining the mechanisms for generating and maintaining this unique neuronal plasma membrane website. The Kv2.1 voltage-gated potassium channel is abundantly present in large clusters within the soma and proximal dendrites of mammalian mind neurons. Kv2.1 is also a component of the ion channel repertoire in the AIS. Here we display that Kv2.1 clusters within the AIS of mind neurons across.