Analysis of neuron anatomy was done as previously described [99 (link)]. Briefly, after filling MSNs with a biocytin-containing intercellular solution during electrophysiological analyses, brain slices were removed from the recording chamber, fixed with 4% Histofix (Roth, Germany) and subsequently stained with Streptavidin-Alexa Fluor 594 (dilution of 1:1000; ThermoFisher Scientific Inc., Waltham, MA, USA). Slices were mounted onto slides with ProLong Gold antifade reagent (ThermoFisher Scientific Inc., Waltham, MA, USA).
Proximal and distal dendrites of MSNs in the striatum were imaged with a 63× objective (Olympus, Shinjuku, Japan) on a confocal microscope (Leica SP5 LSM; Leica, Wetzlar, Germany). Subsequent blind deconvolution was performed with Amira software (ThermoFisher Scientific Inc., Waltham, MA, USA). Whole-cell images for Sholl analysis were taken with a 40× objective (Olympus, Shinjuku, Japan).
Semi-automatic spine counting of proximal and distal dendrites and tracing of dendrites of whole cells was carried out with NeuronStudio (version 0.9.92; Computational Neurobiology and Imaging Center Mount Sinai School of Medicine, New York, NY, USA). Sholl analyses of the reconstructed neurons were performed with Neurolucida (version 3.70.2; MBF Bioscience; Williston, VT, USA), with starting radius and radius increment parameter set to 10 µm.
Proximal and distal dendrites of MSNs in the striatum were imaged with a 63× objective (Olympus, Shinjuku, Japan) on a confocal microscope (Leica SP5 LSM; Leica, Wetzlar, Germany). Subsequent blind deconvolution was performed with Amira software (ThermoFisher Scientific Inc., Waltham, MA, USA). Whole-cell images for Sholl analysis were taken with a 40× objective (Olympus, Shinjuku, Japan).
Semi-automatic spine counting of proximal and distal dendrites and tracing of dendrites of whole cells was carried out with NeuronStudio (version 0.9.92; Computational Neurobiology and Imaging Center Mount Sinai School of Medicine, New York, NY, USA). Sholl analyses of the reconstructed neurons were performed with Neurolucida (version 3.70.2; MBF Bioscience; Williston, VT, USA), with starting radius and radius increment parameter set to 10 µm.
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