Journal: The Journal of Clinical Investigation
Article Title: Regularizing firing patterns of rat subthalamic neurons ameliorates parkinsonian motor deficits
Figure Lengend Snippet: The HCN2 channel is responsible for the histamine-induced amelioration of motor deficits in PD rats. ( A – D ) LV- Hcn1 -shRNA, LV- Hcn2 -shRNA, LV- Hcn3 -shRNA, and LV- Hcn4 -shRNA effectively downregulated the expression of Hcn1 , Hcn2 , Hcn3 , and Hcn4 mRNAs and proteins ( n = 6 from 6 independent experiments) in the STN. ( E ) LV- Hcn2 -oe upregulated the expression of Hcn2 mRNAs and proteins ( n = 6 from 6 independent experiments). ( F – H ) Effects of downregulation and overexpression of the HCN2 channel in the STN on motor deficits of turning behavior ( F , n = 12), adhesive-removal test ( G , n = 10), and locomotor footprints ( H , n = 10) in PD rats with sham operation, saline injection, and histamine injection. Downregulation of the HCN2 channel significantly increased the apomorphine-induced turnings, prolonged contralesional adhesive-removal time, and shortened bilateral stride length, whereas downregulation of the HCN1, HCN3, or HCN4 channel had no effect on these motor deficits. Only the downregulation of HCN2 rather than the HCN1, HCN3, or HCN4 channel blocked the amelioration of turnings, removal time, and stride length of PD rats induced by microinjection of histamine into the STN. Overexpression of the HCN2 channel in STN not only decreased the turnings, reduced removal time, and enlarged bilateral stride length of PD rats, but also improved the histamine-induced amelioration in these motor behaviors. Data are represented as mean ± SEM. *** P
Article Snippet: Sections were incubated overnight at 4°C with primary antibody/antibodies, as follows: mouse anti-glutamate (1:1000, MilliporeSigma; catalog MAB5304, RRID:AB_94698), goat anti-GAD67 (1:1000, Abcam; catalog ab80589, RRID:AB_1640532), rabbit anti-histamine (1:1000, MilliporeSigma; catalog AB5885, RRID:AB_177540) and rabbit anti-histamine (1:500, Acris; catalog 22939, RRID:AB_572245), mouse anti-TH (1:2000, MilliporeSigma; catalog T2928, RRID:AB_477569), goat anti-H1 receptor (1:500, Everest Biotech; catalog EB06904, RRID:AB_2230568) and rabbit anti-H1 receptor (1:50, Santa Cruz Biotechnology Inc.; catalog SC20633, RRID:AB_2277328), goat anti-H2 receptor (1:500, Everest Biotech; catalog EB06905, RRID:AB_2121375), rabbit anti-H4 receptor (1:200, Santa Cruz Biotechnology Inc.; catalog SC50313, RRID:AB_2119026), rabbit anti-HCN1 (1:300, Alomone Labs; catalog APC-056, RRID:AB_2039900), rabbit anti-HCN2 (1:100, Alomone Labs; catalog APC-030, RRID:AB_2313726), rabbit anti-HCN3 (1:100, Alomone Labs; catalog APC-057, RRID:AB_2039904), and rabbit anti-HCN4 (1:200, Alomone Labs; catalog APC-052, RRID:AB_2039906).
Techniques: shRNA, Expressing, Over Expression, Injection