Journal: Science Advances
Article Title: State-dependent modulation of spiny projection neurons controls levodopa-induced dyskinesia in a mouse model of Parkinson’s disease
doi: 10.1126/sciadv.adv8224
Figure Lengend Snippet: ( A ) Experimental timeline. LID mice were euthanized 24 to 48 hours or 30 min after the last levodopa injection for ex vivo assessment of the off- and on-state, respectively. ( B ) Confocal image of a coronal brain slice from a unilaterally 6-OHDA–lesioned, LID on-state mouse immunostained with anti-pERK (red) and anti–tyrosine hydroxylase (TH; green) antibodies. Scale bar, 0.5 mm. ( C ) Schematic illustrating the somatic excitability assay in dSPNs. ( D ) Sample voltage recordings from dSPNs in the off- and on-state of LID in response to a 200-pA current injection. Scale bars, 10 mV and 200 ms. ( E ) Current-response curves showing that somatic excitability of dSPNs was state dependent (control: n = 10 cells from six mice; 6-OHDA: n = 12 cells from 5 mice; off-state: n = 15 cells from 8 mice; on-state: n = 15 cells from 10 mice; on-state + SCH: n = 10 cells from 5 mice). ( F ) Box plot summary of rheobase in dSPNs. **** P < 0.0001; Mann-Whitney test. ctr, control. ( G ) Two-photon laser scanning microscopy (2PLSM) image of a patched dSPN with imaging sites on proximal and distal dendrites indicated. Scale bar, 20 μm. ( H ) Current injections of 2 nA (top) are aligned with evoked somatic spikes (middle) and fluorescence transients recorded from proximal and distal dendrites (bottom). Scale bars, 25 mV, 0.1 ΔG/R 0 , and 0.5 s. ( I ) Box plot summary of dendritic excitability index [the distal-to-proximal ratio of area under the curve (AUC) of ΔG/R 0 ] of dSPNs (control: n = 11 cells from 6 mice; 6-OHDA: n = 9 cells from 5 mice; off-state: n = 10 cells from 7 mice; on-state: n = 10 cells from 5 mice; on-state + SCH: n = 9 cells from 5 mice). * P < 0.05; Mann-Whitney test.
Article Snippet: A two-photon laser (Chameleon Ultra II, Coherent, Santa Clara, CA) tuned to 920 nm was used to excite GRAB ACh3.0 .
Techniques: Injection, Ex Vivo, Slice Preparation, Control, MANN-WHITNEY, Laser-Scanning Microscopy, Imaging, Fluorescence
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![( A ) Experimental timeline. LID mice were euthanized 24 to 48 hours or 30 min after the last levodopa injection for ex vivo assessment of the off- and on-state, respectively. ( B ) Confocal image of a coronal brain slice from a unilaterally 6-OHDA–lesioned, LID on-state mouse immunostained with anti-pERK (red) and anti–tyrosine hydroxylase (TH; green) antibodies. Scale bar, 0.5 mm. ( C ) Schematic illustrating the somatic excitability assay in dSPNs. ( D ) Sample voltage recordings from dSPNs in the off- and on-state of LID in response to a 200-pA current injection. Scale bars, 10 mV and 200 ms. ( E ) Current-response curves showing that somatic excitability of dSPNs was state dependent (control: n = 10 cells from six mice; 6-OHDA: n = 12 cells from 5 mice; off-state: n = 15 cells from 8 mice; on-state: n = 15 cells from 10 mice; on-state + SCH: n = 10 cells from 5 mice). ( F ) Box plot summary of rheobase in dSPNs. **** P < 0.0001; Mann-Whitney test. ctr, control. ( G <t>)</t> <t>Two-photon</t> laser scanning microscopy (2PLSM) image of a patched dSPN with imaging sites on proximal and distal dendrites indicated. Scale bar, 20 μm. ( H ) Current injections of 2 nA (top) are aligned with evoked somatic spikes (middle) and fluorescence transients recorded from proximal and distal dendrites (bottom). Scale bars, 25 mV, 0.1 ΔG/R 0 , and 0.5 s. ( I ) Box plot summary of dendritic excitability index [the distal-to-proximal ratio of area under the curve (AUC) of ΔG/R 0 ] of dSPNs (control: n = 11 cells from 6 mice; 6-OHDA: n = 9 cells from 5 mice; off-state: n = 10 cells from 7 mice; on-state: n = 10 cells from 5 mice; on-state + SCH: n = 9 cells from 5 mice). * P < 0.05; Mann-Whitney test.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4127/pmc12674127/pmc12674127__sciadv.adv8224-f1.jpg)