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. 2022 Apr 25;3:861598. doi: 10.3389/fpain.2022.861598

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Copyright © 2022 Wang, Duong, Rea, Waite, Huebner, Flinn, Russo and Sowers.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The diffusion range of CGRP. (A) Representative example of a mouse after injection of fluorescein-15-CGRP. Upper panel: In the most rostral section, dim fluorescence was detected in the inferior colliculus, the parabrachial nucleus, vermal lobules II-V, and the simple lobule. Middle panel: Fluorescein-15-CGRP at the injection site. Areas within rectangle are magnified in boxes 1 and 2. Clusters of fluorescein-15-CGRP were detected in cell bodies in the MN (box 1) and nearby cells, including the interposed and lateral cerebellar nuclei, granular, Purkinje cell, and molecular layers of vermal lobules I/III/IV/V (box 2). Dim signal was found in the simple lobule of the hemispheric regions. Lower panel: In the most caudal section, dim fluorescence was detected in lobule IX. Green: fluorescein-15-CGRP; Blue: TO-PRO-3. (B) The spread of the green fluorescence among the mice injected with fluorescein-15-CGRP. The smallest (purple shading) spread of signals covers the MN and few of nearby cells in the vermal lobules III/IV/V. The largest spread (blue shading) covers the MN and cells beyond the MN including vermal lobules I/III/IV/V/X, the simple lobule and other cerebellar deep nuclei. In summary, florescent signals were found in vermal lobules I-X, the simple lobule in the hemispheric region, and the midbrain (mainly in superior and inferior colliculus) from rostrally to caudally. (C) A representative image of a mouse with injection of Evans blue. (D) A representative image of a mouse with injection of red beads. DN, lateral cerebellar nucleus (dentate nucleus); GL, granular layer; IC, inferior colliculus; IP, interposed nucleus; ML, molecular layer; MN, medial cerebellar nucleus; PB, parabrachial nucleus; PCL, Purkinje cell layer; SIM, simple lobule. Image credit: Allen Institute. Numbers indicate the distance from bregma in the anteroposterior plane in Allen Mouse Brain Atlas coronal images.