Journal: bioRxiv

Article Title: Identifying the Brain Circuits that Regulate Pain-Induced Sleep Disturbances

doi: 10.1101/2024.12.20.629596

Figure Lengend Snippet: Pharmacological blocking of CGRP or NMDA receptors in the SI-BF or CeA during AIP-induced sleep loss: The experimental protocol for recording sleep with or without AIP and after blocking of either the CGRP (BIBN4096BS; 80µg/ µl) or NMDA (AP-5; 40µg/ µl) receptors with 200nl of antagonists injected bilaterally in either the SI- BF or the CeA using the injectors cannulas and the infusion/withdrawal programmable dual syringe pump ( a ). Photomicrographs of Nissl’s-stained brain sections ( b ) show the guide-cannula tracks used to insert the injector cannulas, whose tips were marked using thionine stain, in the vicinity (within a radius of 1-1.5mm) of the substantia innominata (SI, b1 ) in the basal forebrain (BF) and the central nucleus of the amygdala (CeA, b2 ). Asterisks mark the tip of the injector cannula that extended the guide cannulas by 1 mm, observed either in the same or adjacent section that lie within 1mm of the PBel CGRP terminal sites. Scale in b1 and b2 is 0.5mm. Graphs in c show the percentage (mean ± SEM) of time spent in NREM sleep for each hour ( c1) and in 3h bins ( c2 ) after AIP. The blue rectangle represents the dark phase. The groups were statistically compared using a two-way (treatment X time) ANOVA, followed by Holms-sidak method for multiple comparisons, where **- P<0.001; *- P<0.01. The color of the asterisk compares AIP to the group representing the color in the graph.

Article Snippet: For the pharmacological blocking, we used CGRP receptor blocker injections, BIBN (BIBN4096BS; 80μg/ml; Boehringer Ingelheim Pharma KG, Biberach, Germany) and NMDA receptor blocker injection, AP-5 (40μg/ml), which were delivered using the injector cannula that was placed in the guide and extending past the tip of the guide by 1mm.

Techniques: Blocking Assay, Injection, Staining