Comparison of the expression of neurotransmitter and muscular genesis markers in the postnatal male mouse masseter and trigeminal ganglion during development.

Calcitonin gene-related peptide (CGRP) is released by motor neurons and affects skeletal muscle fiber and transient receptor potential cation channel subfamily V member 1 (TRPV1), an important marker of pain modulation. However, the expression of CGRP and TRPV1 in the trigeminal ganglion (TG) during changes and in feeding patterns has not been described. We used real-time reverse transcription polymerase chain reaction and in situ hybridization to investigate the mRNA expression levels of CGRP and TRPV1 in the TG. The expression of myosin heavy-chain (MyHC) isoforms was also investigated in the masseter muscle (MM) during the transition from sucking to mastication, an important functional trigger for muscle. The mRNA and protein levels of CGRP increased in the MM and TG from postnatal day 10 (P10) to P20 in male mice. The protein levels of TRPV1 were almost constant in the TG from P10 to P20, in contrast to increases in the MM. The mRNA abundance of TRPV1 in the TG and MM was increased from P10 to P20. The localization of an antisense probe was used to count CGRP cell numbers and found to differentiate the ophthalmic, maxillary, and mandibular nerve divisions of the TG. In particular, the number of CGRP+ cells per 10,000 µm2 in the maxillary and mandibular divisions of the TG gradually changed from P10 to P20. The expression of CGRP and TRPV1 in the TG and MM and the patterns of expression of different MyHC isoforms were affected by changes in feeding during male mouse development.

Distribution of glutamate receptor, ionotropic, kainate 1 and neuropeptide calcitonin gene-related peptide mRNAs during formation of the embryonic and postnatal mouse molar in the maxilla.

The neuropeptide calcitonin gene-related peptide (CGRP) is a well-characterized neurotransmitter. Glutamate receptor, ionotropic, kainate 1 (Grik1) has also been demonstrated to generate high-affinity kainate receptors. However, little is known about the roles of CGRP and Grik1 during the developmental formation of teeth. In this study, we endeavoured to analyse the expression and localization of CGRP and Grik1 mRNAs using in situ hybridization on the mouse maxilla during development from the embryonic stage (E18.5) to after birth (P10, P15 and P20). We found that hybridization with an anti-sense probe for CGRP clearly localized in the maxilla at E18.5 in contrast to that of P15 and P20. Hybridization with an anti-sense probe for CGRP was not detected in the dental pulp of molars in the maxilla at P10, which is in contrast to Grik1 mRNA at the same developmental stage. Hybridization with an anti-sense probe for Grik1 mRNA was detected in the basal region of the dental pulp of molars at P10 and P15. Finally, these markers were not detected in molars in the mouse maxilla at P20. The ratio of positive cells for the hybridization signals of Grik1and CGRP in the dental pulp decreased from E18.5 (p<0.001). These features in CGRP and Grik1r mRNAs may indicate roles of function during tooth development between embryonic and postnatal stages with root formation and erupted movements.

Local administration of resveratrol inhibits excitability of nociceptive wide-dynamic range neurons in rat trigeminal spinal nucleus caudalis.

Although we recently reported that intravenous administration of resveratrol suppresses trigeminal nociception, the precise peripheral effect of resveratrol on nociceptive and non-nociceptive mechanical stimulation-induced trigeminal neuron activity in vivo remains to be determined. The aim of the present study was to investigate whether local subcutaneous administration of resveratrol attenuates mechanical stimulation-induced excitability of trigeminal spinal nucleus caudalis (SpVc) neuron activity in rats, in vivo. Extracellular single-unit recordings were made of SpVc wide-dynamic range (WDR) neuron activity in response to orofacial mechanical stimulation in pentobarbital-anesthetized rats. Neurons responded to non-noxious and noxious mechanical stimulation applied to the orofacial skin. Local subcutaneous administration of resveratrol (1-10mM) into the orofacial skin dose dependently and significantly reduced the mean number of SpVc WDR neurons firing in response to both non-noxious and noxious mechanical stimuli, with the maximal inhibition of discharge frequency in response to both stimuli being seen within 5min. These inhibitory effects were no longer evident after approximately 20min. The mean magnitude of inhibition by resveratrol (10mM) of SpVc neuron discharge frequency was almost equal to that of the local anesthetic 1% lidocaine (37mM). These results suggest that local injection of resveratrol into the peripheral receptive field suppresses the excitability of SpVc neurons, possibly via inhibition of Na(+) channels in the nociceptive nerve terminals of trigeminal ganglion neurons. Therefore, local subcutaneous administration of resveratrol may provide relief of trigeminal nociceptive pain, without side effects, thus contributing to the suite of complementary and alternative medicines used as local anesthetic agents.