A truncated Kv1.1 protein in the brain of the mouse: expression and interaction-2

<p><b>Copyright information:</b></p><p>Taken from "A truncated Kv1.1 protein in the brain of the mouse: expression and interaction"</p><p>BMC Neuroscience 2005;6():65-65.</p><p>Published online 23 Nov 2005</p><p>PMCID:PMC1322225.</p><p>Copyright © 2005 Persson et al; licensee BioMed Central Ltd.</p>ern. Both EndoH and PNGaseF reduced the molecular weight with approximately 3 kDa (arrow). This corresponds to core glycosylation. No unglycosylated MCEPH was detected

A truncated Kv1.1 protein in the brain of the mouse: expression and interaction-1

<p><b>Copyright information:</b></p><p>Taken from "A truncated Kv1.1 protein in the brain of the mouse: expression and interaction"</p><p>BMC Neuroscience 2005;6():65-65.</p><p>Published online 23 Nov 2005</p><p>PMCID:PMC1322225.</p><p>Copyright © 2005 Persson et al; licensee BioMed Central Ltd.</p>adjusted to obtain a strong and clear signal. Note exposure times given to relate between panels A. Wild type hippocampus showed the same staining pattern as that previously reported for Kv1.1 [2] (exposure 1100 ms) B. Kv1.1 null mouse hippocampus: some cross reactivity of the antibody was seen. (exposure 2700 ms) C. In the hippocampus the immunoreactivity surrounded the nuclei of neurons especially in the dentate gyrus hilus (h) (exposure 2500 ms) D. Parietal neocortex in (left) and wild type (right) brain: the staining of fibers in wild type was absent in . Scale bar 200 μm; h, dentate gyrus hilus; Gr, dentate gyrus granular cell layer; wt, wild type; -/-, Kv1.1-null; m/m

A truncated Kv1.1 protein in the brain of the mouse: expression and interaction-3

<p><b>Copyright information:</b></p><p>Taken from "A truncated Kv1.1 protein in the brain of the mouse: expression and interaction"</p><p>BMC Neuroscience 2005;6():65-65.</p><p>Published online 23 Nov 2005</p><p>PMCID:PMC1322225.</p><p>Copyright © 2005 Persson et al; licensee BioMed Central Ltd.</p>y on brain lysate from wild type (+/+) and Kv1.1 null (-/-) mice. In wild type brain both Kv1.1 and Kv1.2 are detected. In Kv1.1 null brain neither Kv1.1 nor Kv1.2 was detected. B. Immunoprecipitation was performed with the anti-Kv1.2 monoclonal antibody on brain lysate from wild type (+/+) and (m/m) mice. The immunoprecipitation reaction and corresponding brain lysate was loaded on SDS-PAGE. In brain lysate from (BL) MCEPH was detected using the polyclonal Kv1.1 N-terminal antibody (arrow). However, no MCEPH band was detected in the immunoprecipitate from (IP). C. Hippocampi were dissected and an equal amount of lysate from wild type and was loaded on SDS-PAGE and immunoblotted with anti-Kv1.2. Only a very small fraction of the Kv1.2 was core glycosylated (60 kDa). There appeared to be no increase in the amount of core glycosylated Kv1.2 in hippocampus compared to wild type. D. HEK293 cells were cotransfected with Kv1.1-DsRed and MCEPH-ZsGreen constructs. Both the 85 kDa Kv1.1-DsRed and the 55 kDa MCEPH-ZsGreen fusion proteins were detected with immunoblotting on cell lysate using the polyclonal Kv1.1 N-terminal antibody (lysate). The relative levels of the two fusion proteins in this overepressing cell system cannot be used to quantitate MCEPH expression or stability in brain since regulation and trafficking is known to be different between these two systems. The lysate was immunoprecipitated with the Kv1.1 C-terminal antibody. In the precipitate, both fusion proteins were detected with the Kv1.1 N-terminal antibody (IP). When the Kv1.1 C-terminal antibody was used for immunoblotting only the Kv1.1-DsRed fusion protein was detected

A truncated Kv1.1 protein in the brain of the mouse: expression and interaction-0

<p><b>Copyright information:</b></p><p>Taken from "A truncated Kv1.1 protein in the brain of the mouse: expression and interaction"</p><p>BMC Neuroscience 2005;6():65-65.</p><p>Published online 23 Nov 2005</p><p>PMCID:PMC1322225.</p><p>Copyright © 2005 Persson et al; licensee BioMed Central Ltd.</p> SDS-PAGE and immunoblotted with the polyclonal Kv1.1 N-terminal antibody. In wild type lysate a strong band was detected at 86 kDa, corresponding to Kv1.1. The same band was seen in the Kv1.1 null and lysates but at a lower intensity. The 86 kDa bands in Kv1.1 null and lysates are due to antibody cross reactivity since neither Kv1.1 null nor mice have any full-length Kv1.1 protein. B. The polyclonal Kv1.1 N-terminal antibody was preincubated with the peptide used for immunization. Lysate from wild type (+/+) and (m/m) brains were loaded on SDS-PAGE and immunoblotted with the Kv1.1 N-terminal antibody without or after preincubation. The preincubation completely blocked the signal. C. A longer exposure of the immunoblot in panel A. In brain lysate there was a unique band at approximately 30 kDa, which corresponds to the calculated weight of MCEPH (arrow)

Highly Selective Dopamine D₃ Receptor Antagonists with Arylated Diazaspiro Alkane Cores

A series of potent and selective D₃ receptor (D₃R) analogues with diazaspiro alkane cores were synthesized. Radioligand binding of compounds <b>11</b>, <b>14</b>, <b>15a</b>, and <b>15c</b> revealed favorable D₃R affinity (<i>K</i>_i = 12–25.6 nM) and were highly selective for D₃R vs D₃R (ranging from 264- to 905-fold). Variation of these novel ligand architectures can be achieved using our previously reported 10–20 min benchtop C–N cross-coupling methodology, affording a broad range of arylated diazaspiro precursors

Highly Selective Dopamine D₃ Receptor Antagonists with Arylated Diazaspiro Alkane Cores

A series of potent and selective D₃ receptor (D₃R) analogues with diazaspiro alkane cores were synthesized. Radioligand binding of compounds <b>11</b>, <b>14</b>, <b>15a</b>, and <b>15c</b> revealed favorable D₃R affinity (<i>K</i>_i = 12–25.6 nM) and were highly selective for D₃R vs D₃R (ranging from 264- to 905-fold). Variation of these novel ligand architectures can be achieved using our previously reported 10–20 min benchtop C–N cross-coupling methodology, affording a broad range of arylated diazaspiro precursors