5HTR transcripts coexpress with TRPV1. 5HT increases calcium accumulation and CGRP release from cultured capsaicin-sensitive trigeminal cells, which is attenuated with 5HTR antagonists and sumatriptan.
Peripheral serotonin (5HT) has been implicated in migraine and temporomandibular pain disorders in humans and animal models and yet the mechanism(s) by which 5HT evokes pain remains unclear. Trigeminal pain can be triggered by activation of the transient receptor potential V1 channel (TRPV1), expressed by a subset of nociceptive trigeminal ganglia (TG) neurons and gated by capsaicin, noxious heat, and other noxious stimuli. As 5HT is released in the periphery during inflammation and evokes thermal hyperalgesia, and TRPV1 is essential for thermal hyperalgesia, we hypothesized that 5HT increases the activity of capsaicin-sensitive trigeminal neurons and that this increase can be attenuated by pharmacologically targeting peripheral 5HT receptors. TG cultures were pretreated with 5HT (10 nM–100 μM), sumatriptan (5HT1B/1D agonist), ketanserin (5HT2A antagonist), granisetron (5HT3 antagonist), or vehicle prior to capsaicin (30–50 nM). Single-cell accumulation of intracellular calcium was recorded or calcitonin gene-related peptide (CGRP) release was measured following each treatment. In addition, using in situ hybridization and immunohistochemistry, we detected the colocalization of 5HT1B, 5HT1D, 5HT2A, and 5HT3A, but not 5HT2C mRNA with TRPV1 in TG cells. 5HT pretreatment evoked a significant increase in calcium accumulation in capsaicin-sensitive trigeminal neurons and enhanced capsaicin-evoked CGRP release, but had no significant effect when given alone. Sumatriptan, ketanserin, and granisetron treatment attenuated calcium accumulation and 5HT enhancement of capsaicin-evoked CGRP release. Together these results indicate that 5HT increases the activity of capsaicin-sensitive peripheral nociceptors, which can be attenuated by pharmacologically targeting peripheral 5HT receptors, thereby providing a mechanistic basis for peripheral craniofacial pain therapy.
aDepartment of Endodontics, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA
bDepartment of Pharmacology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA
*Corresponding author. Address: Department of Endodontics, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229–3900, USA. Tel.: +1 210 567 0779; fax: +1 210 567 3389.
Submitted December 10, 2010; revised June 1, 2011; accepted June 6, 2011.