Watanabe Masaya, Ueda Takashi, Shibata Yasuhiro, Kumamoto Natsuko, Shimada Shoichi, Ugawa Shinya
Department of Anatomy and Neuroscience, Graduate School of Medical Sciences, Nagoya City University, Nagoya, Japan.
Department of Neuroscience and Cell Biology, Osaka University Graduate School of Medicine, Suita, Japan.
PLoS One. 2015 May 14;10(5):e0127572. doi: 10.1371/journal.pone.0127572. eCollection 2015.
The Cav3.2 isoform of the T-type calcium channel is expressed in primary sensory neurons of the dorsal root ganglion (DRG), and these channels contribute to nociceptive and neuropathic pain in rats. However, there are conflicting reports on the roles of these channels in pain processing in rats and mice. In addition, the function of T-type channels in persistent inflammatory hyperalgesia is poorly understood. We performed behavioral and comprehensive histochemical analyses to characterize Cav3.2-expressing DRG neurons and examined the regulation of T-type channels in DRGs from C57BL/6 mice with carrageenan-induced inflammatory hyperalgesia. We show that approximately 20% of mouse DRG neurons express Cav3.2 mRNA and protein. The size of the majority of Cav3.2-positive DRG neurons (69 ± 8%) ranged from 300 to 700 μm2 in cross-sectional area and 20 to 30 μm in estimated diameter. These channels co-localized with either neurofilament-H (NF-H) or peripherin. The peripherin-positive cells also overlapped with neurons that were positive for isolectin B4 (IB4) and calcitonin gene-related peptide (CGRP) but were distinct from transient receptor potential vanilloid 1 (TRPV1)-positive neurons during normal mouse states. In mice with carrageenan-induced inflammatory hyperalgesia, Cav3.2 channels, but not Cav3.1 or Cav3.3 channels, were upregulated in ipsilateral DRG neurons during the sub-acute phase. The increased Cav3.2 expression partially resulted from an increased number of Cav3.2-immunoreactive neurons; this increase in number was particularly significant for TRPV1-positive neurons. Finally, preceding and periodic intraplantar treatment with the T-type calcium channel blockers mibefradil and NNC 55-0396 markedly reduced and reversed mechanical hyperalgesia during the acute and sub-acute phases, respectively, in mice. These data suggest that Cav3.2 T-type channels participate in the development of inflammatory hyperalgesia, and this channel might play an even greater role in the sub-acute phase of inflammatory pain due to increased co-localization with TRPV1 receptors compared with that in the normal state.
T型钙通道的Cav3.2亚型在背根神经节(DRG)的初级感觉神经元中表达,并且这些通道在大鼠的伤害性疼痛和神经性疼痛中起作用。然而,关于这些通道在大鼠和小鼠疼痛处理中的作用存在相互矛盾的报道。此外,T型通道在持续性炎症性痛觉过敏中的功能尚不清楚。我们进行了行为学和全面的组织化学分析以表征表达Cav3.2的DRG神经元,并研究了角叉菜胶诱导的炎症性痛觉过敏的C57BL/6小鼠DRG中T型通道的调节。我们发现约20%的小鼠DRG神经元表达Cav3.2 mRNA和蛋白。大多数Cav3.2阳性DRG神经元(69±8%)的大小,横截面积范围为300至700μm²,估计直径为20至30μm。这些通道与神经丝-H(NF-H)或外周蛋白共定位。外周蛋白阳性细胞也与isolectin B4(IB4)和降钙素基因相关肽(CGRP)阳性的神经元重叠,但在正常小鼠状态下与瞬时受体电位香草酸亚型1(TRPV1)阳性神经元不同。在角叉菜胶诱导的炎症性痛觉过敏小鼠中,在亚急性期,同侧DRG神经元中Cav3.2通道上调,但Cav3.1或Cav3.3通道未上调。Cav3.2表达增加部分是由于Cav3.2免疫反应性神经元数量增加;这种数量增加对于TRPV1阳性神经元尤为显著。最后,在小鼠中,预先和定期足底内注射T型钙通道阻滞剂米贝拉地尔和NNC 55-0396分别在急性期和亚急性期显著减轻并逆转了机械性痛觉过敏。这些数据表明Cav3.2 T型通道参与炎症性痛觉过敏的发展,并且与正常状态相比,由于与TRPV1受体的共定位增加,该通道在炎症性疼痛的亚急性期可能发挥更大作用。