Cell-attached recordings of CIN activity were performed in male C57 mice (~30-60 days old, Jackson Laboratory) with glass micropipettes (2–4 MΩ) filled with aCSF. Cholinergic interneurons were identified based on their large size, tonic firing, sensitivity to muscarine, and sensitivity to Sk blockade (Yorgason et al., 2017 (link)). Data were filtered at the amplifier with a band-pass Bessel filter 0.3-3 kHz (Axon Instruments Multiclamp 700A; Molecular Devices, San Jose CA). Recordings were sampled continuously at 10 kHz with a PCIe-6321 NI-DAQ board (National Instruments, Austin, TX) and acquired using Axograph 1.7.6 (Axograph Scientific). Recordings were obtained over a 10 minute period, and the last 3 minutes of recordings were used for analysis. The firing rate was analyzed using a simple threshold detection algorithm in Axograph, and focused on measures of instantaneous firing frequency. Values reported are average instantaneous firing frequency over a 3 minute period.
Glass micropipettes
Manufactured by Jackson ImmunoResearch
Glass micropipettes are precision laboratory instruments used to accurately measure and transfer small volumes of liquids. These pipettes are typically made of borosilicate glass and are designed to deliver precise amounts of fluids ranging from microliters to milliliters, with high accuracy and repeatability.
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2 protocols using glass micropipettes
1
Electrophysiological Recording of Cholinergic Interneurons
2
Electrophysiological Recording of Cholinergic Interneurons
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Cell-attached recordings of CIN activity were performed in male C57 mice (~30-60 days old, Jackson Laboratory) with glass micropipettes (2–4 MΩ) filled with aCSF. Cholinergic interneurons were identified based on their large size, tonic firing, sensitivity to muscarine, and sensitivity to Sk blockade (Yorgason et al., 2017 (link)). Data were filtered at the amplifier with a band-pass Bessel filter 0.3-3 kHz (Axon Instruments Multiclamp 700A; Molecular Devices, San Jose CA). Recordings were sampled continuously at 10 kHz with a PCIe-6321 NI-DAQ board (National Instruments, Austin, TX) and acquired using Axograph 1.7.6 (Axograph Scientific). Recordings were obtained over a 10 minute period, and the last 3 minutes of recordings were used for analysis. The firing rate was analyzed using a simple threshold detection algorithm in Axograph, and focused on measures of instantaneous firing frequency. Values reported are average instantaneous firing frequency over a 3 minute period.
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