Meningeal lymphatic CGRP signaling governs pain via cerebrospinal fluid efflux and neuroinflammation in migraine models
Nathan P. Nelson-Maney, László Bálint, Anna L.S. Beeson, D. Stephen Serafin, Bryan M. Kistner, Elizabeth S. Douglas, Aisha H. Siddiqui, Alyssa M. Tauro, Kathleen M. Caron
Nathan P. Nelson-Maney, László Bálint, Anna L.S. Beeson, D. Stephen Serafin, Bryan M. Kistner, Elizabeth S. Douglas, Aisha H. Siddiqui, Alyssa M. Tauro, Kathleen M. Caron
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Research Article Cell biology Vascular biology

Meningeal lymphatic CGRP signaling governs pain via cerebrospinal fluid efflux and neuroinflammation in migraine models

Abstract

Recently developed antimigraine therapeutics targeting calcitonin gene–related peptide (CGRP) signaling are effective, though their sites of activity remain elusive. Notably, the lymphatic vasculature is responsive to CGRP signaling, but whether meningeal lymphatic vessels (MLVs) contribute to migraine pathophysiology is unknown. Mice with lymphatic vasculature deficient in the CGRP receptor (CalcrliLEC mice) treated with nitroglycerin-mediated (NTG-mediated) chronic migraine exhibit reduced pain and light avoidance compared with NTG-treated littermate controls. Gene expression profiles of lymphatic endothelial cells (LECs) isolated from the meninges of Rpl22HA/+;Lyve1Cre RiboTag mice treated with NTG revealed increased MLV-immune interactions compared with cells from untreated mice. Interestingly, the relative abundance of mucosal vascular addressin cell adhesion molecule 1–interacting (MAdCAM1-interacting) CD4+ T cells was increased in the deep cervical lymph nodes of NTG-treated control mice but not in NTG-treated CalcrliLEC mice. Treatment of cultured hLECs with CGRP peptide in vitro induced vascular endothelial–cadherin (VE-cadherin) rearrangement and reduced functional permeability. Likewise, intra cisterna magna injection of CGRP caused rearrangement of VE-cadherin, decreased MLV uptake of cerebrospinal fluid (CSF), and impaired CSF drainage in control mice but not in CalcrliLEC mice. Collectively, these findings reveal a previously unrecognized role for lymphatics in chronic migraine, whereby CGRP signaling primes MLV-immune interactions and reduces CSF efflux.

Authors

Nathan P. Nelson-Maney, László Bálint, Anna L.S. Beeson, D. Stephen Serafin, Bryan M. Kistner, Elizabeth S. Douglas, Aisha H. Siddiqui, Alyssa M. Tauro, Kathleen M. Caron

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Figure 2

CalcrliLEC mice treated with NTG exhibit ameliorated light avoidance and anxiety behavior.

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CalcrliLEC mice treated with NTG exhibit ameliorated light avoidance an...
(A) Schematic of light aversion and movement assay. (B) Schematic of general (top) and daily (bottom) experimental protocol. Mice were allowed to acclimate to the chamber 4 days before the first injection and 2 days before the baseline measurement (baseline measurement recorded on day –1) Light avoidance and movement behavior are measured 30 minutes after NTG injection and were recorded for 30 minutes. (C) Time spent in dark chamber reported for all test days and on day 1 (D) and day 5 (E). Dashed line shows the total test time (1,800 seconds or 30 minutes). (F) Time spent in light chamber reported for all test days. (G) Normalized percent time spent moving in the dark chamber and (H) normalized percent time spent in moving in the light chamber for all experimental days. Percent of time spent moving in the respective chamber were moving normalized to baseline movement in the same chamber data. Data are normalized to the baseline measurement on day –1 by individual calculated as percent time moving on test day divided by baseline percent time moving (day –1) multiplied by 100. *P < 0.05, **P < 0.01 reported between Calcrlfl/fl and CalcrliLEC treated with NTG. Dashed line, baseline. C–H, n = 14–20 animals per group. Data represents 12 independent cohorts. Significance for all data calculated using 2-way ANOVA with Tukey’s multiple comparisons test. P values indicated if < 0.1. Graphs = mean ± SD.