Journal: eLife
Article Title: Chronic hyperactivation of midbrain dopamine neurons causes preferential dopamine neuron degeneration
doi: 10.7554/eLife.98775
Figure Lengend Snippet: ( A ) Graphical illustration summarizing experimental design. Recombinant AAV encoding a conditional allele of the hM3Dq(DREADD)-mCherry was injected bilaterally into the ventral midbrain of 4- to 5-month-old DAT IRES cre mice. CNO (300 mg/L) or vehicle (2% sucrose in water) was administered ad libitum via drinking water for 2 weeks, and the animals were perfused the next day. Changes in locomotion were assessed with running wheels. The 2 days preceding the start of treatment were used as a measure for baseline locomotion. ( B ) Representative traces of wheel usage for animals given control vehicle water (top) or CNO water (bottom). Arrows denote start of treatment. Gray background shading indicates dark cycle hours. ( C ) Mean wheel usage for selected days during the experiment, segregated by light (left), dark (middle), and total (light+dark; right) cycles. n=10 animals/group from 2 independent experiments. ( D ) Spontaneous firing rate was measured during the first 2 min of whole-cell recordings. n = 21 neurons from 4 vehicle-treated mice and 18 neurons from 5 CNO-treated mice for SN and 11 neurons from 2 vehicle-treated mice and 19 neurons from 2 CNO-treated mice for VTA. ( E ) Time course of responses to bath application of 1 μM CNO ex vivo measured in current clamp in neurons from vehicle-treated and CNO-treated mice. n = 5 neurons from vehicle-treated mice and 9 neurons from CNO-treated mice for SN and 3 neurons from vehicle-treated mice and 7 neurons from CNO-treated mice. Data indicate mean ± SEM. *p≤0.05, **p<0.01, ***p<0.001 by two-way ANOVA and Holm-Sidak post hoc test ( C ). *p≤0.05, **p=0.01 by t-test or permutation (nonparametric) analysis ( D ).
Article Snippet: Two weeks following surgery, animals were single-housed and habituated to wireless running wheels (MedAssociates ENV-047, RRID: SCR_024879 ) connected to a hub (MedAssociates DIG-807, RRID: SCR_024880 ) for locomotion recordings and water bottles (Amazon) for drinking.
Techniques: Recombinant, Injection, Control, Ex Vivo
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