A novel technique of delivering viral vectors to trigeminal ganglia in rats

Abstract

The objective of this study was to develop a viable and reliable technique of delivering viral vectors to rat trigeminal ganglia. Adult Sprague-Dawley rats (200-300 g) were used, and lentiviral vectors containing enhanced green fluorescence protein and calcitonin gene-related peptide short hairpin RNA (shRNA) were generated. Following general anesthesia, viral vectors were delivered to rat trigeminal ganglia using the technique described in this study. Both X-ray and micro-computed tomography (micro-CT) were employed to verify the position of the needles when injecting the vectors. In vivo fluorescence imaging and immunostaining against enhanced green fluorescence protein were performed to determine the success of viral transduction.The levels of calcitonin gene-related peptide in trigeminal ganglia were determined using real-time PCR, and pain levels following injections were evaluated using the Rat Grimace Scale. Our results show that injection needles can be advanced precisely at the trigeminal fossa and that viral vectors can successfully transduce trigeminal ganglia. Moreover, the levels of calcitonin gene-related peptide at trigeminal ganglia were down-regulated on day 7 after viral transduction. Pain levels returned to baseline by day 7 following injection. Therefore, we suggest that our trigeminal ganglion-targeting technique could be used for delivering genes or drugs to rat trigeminal ganglia.

Migraine-Associated TRESK Mutations Increase Neuronal Excitability through Alternative Translation Initiation and Inhibition of TREK

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Effect of endomorphin‐2 on orofacial pain induced by orthodontic tooth movement in rats

“…Based on our previous study , sample‐size calculation was based on a difference in bite force of 10.9 ± 3.0 (mean±SD) and was performed using statsdirect 2.7.9 (StatsDirect, Cheshire, UK). The results indicated that a minimum of three rats in each group at each time point were required.…”

“…The technique of trigeminal injection has been described in our previous study . In brief, following inhalation anesthesia with sevoflurane, the hair between the left ears and eyes of rats was rapidly removed with clippers.…”

“…Currently, several experimental methods have been developed for orofacial pain assessment in rodents, such as facial expressions and facial grooming . In our past research, orthodontic pain levels in rats were evaluated through facial expression analysis.…”

N/OFQ modulates orofacial pain induced by tooth movement through CGRP-dependent pathways

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