Lumbar splanchnic nerve (LSN) afferent preparations were conducted as previously described (47 (link)). Briefly, the distal colorectum (splenic flexure to rectum) and associated LSN (rostral to inferior mesenteric ganglia) were isolated from mice euthanized as described earlier. The colon was cannulated with fine thread (polyester, Gutermann) in a rectangular recording chamber with Sylgard base (Dow Corning, UK) and serosally superfused (7 mL/min; 32°C–34°C) and luminally perfused (200 μL/min) by a syringe pump (Harvard apparatus, MA) with carbogenated Krebs buffer solution (95% O2–5% CO2). Krebs buffer was supplemented with 10 μM atropine and 10 μM nifedipine to prevent smooth muscle activity (49 (link)).
Borosilicate suction electrodes were used to record the multiunit activity of LSN bundles. Signals were amplified (gain 5 kHz), band pass filtered (100–1,300 Hz; Neurolog, Digitimer Ltd, UK), and digitally filtered for 50 Hz noise (Humbug, Quest Scientific, Canada). Analog signals were digitized at 20 kHz (Micro1401; Cambridge Electronic Design, UK), and signals were visualized with Spike2 software (Cambridge Electronic Design, UK).
Borosilicate suction electrodes were used to record the multiunit activity of LSN bundles. Signals were amplified (gain 5 kHz), band pass filtered (100–1,300 Hz; Neurolog, Digitimer Ltd, UK), and digitally filtered for 50 Hz noise (Humbug, Quest Scientific, Canada). Analog signals were digitized at 20 kHz (Micro1401; Cambridge Electronic Design, UK), and signals were visualized with Spike2 software (Cambridge Electronic Design, UK).
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