Department of Neurosurgery, Beijing Sanbo Brain Hospital, Capital Medical University, Beijing 100093, P.R. China.
Department of Neurosurgery, The First Hospital of China Medical University, Shenyang 110001, P.R. China.
Int J Mol Med. 2018 Apr;41(4):2225-2232. doi: 10.3892/ijmm.2018.3446. Epub 2018 Jan 30.
Previous studies demonstrated that Nav1.5 splice variants, including Nav1.5a and Nav1.5c, were expressed in dorsal root ganglia (DRG) neurons. However, since nine Nav1.5 isoforms have been identified, whether other Nav1.5 splice variants, especially the neonatal Nav1.5 splice variant, express in the DRG neurons is still unknown. In this study, we systematically investigated the expression of adult and neonatal Nav1.5 isoforms in the DRG neurons and axon of peripheral sensory neurons of rats with spared nerve injury (SNI) by RT-PCR, DNA sequencing, restriction enzyme digestion, immunohistochemistry and immunofluorescence methods. The results demonstrated that both adult and neonatal Nav1.5 isoforms were expressed in the DRG neurons, but their expression ratio was ~2.5:1. In SNI rat models, the expression of both adult and neonatal Nav1.5 decreased by approximately a half in both mRNA and protein levels. In contrast, the expression of protein kinase C (PKC)-γ, one of the negative modulators for sodium currents, increased by ~1-fold. Taken together, this study first confirmed the expression of both adult and neonatal Nav1.5 isoforms in the DRG neurons and axon of peripheral sensory neurons of rat, but their expression level decreased in pain models. The upregulation of PKC-γ may directly or indirectly downregulate the expression of Nav1.5 isoforms in SNI rat models, which may further involve in the pathological process of neuropathic pain.
先前的研究表明,Nav1.5 剪接变异体,包括 Nav1.5a 和 Nav1.5c,在背根神经节(DRG)神经元中表达。然而,由于已经鉴定出了 9 种 Nav1.5 同工型,其他 Nav1.5 剪接变异体,特别是新生 Nav1.5 剪接变异体,是否在 DRG 神经元中表达仍然未知。在这项研究中,我们通过 RT-PCR、DNA 测序、限制性内切酶消化、免疫组织化学和免疫荧光方法,系统地研究了成年和新生 Nav1.5 同工型在 spared nerve injury(SNI)大鼠的 DRG 神经元和周围感觉神经元轴突中的表达。结果表明,成年和新生 Nav1.5 同工型都在 DRG 神经元中表达,但它们的表达比例约为 2.5:1。在 SNI 大鼠模型中,成年和新生 Nav1.5 的表达在 mRNA 和蛋白水平上都减少了约一半。相比之下,蛋白激酶 C(PKC)-γ的表达增加了约 1 倍,PKC-γ 是钠离子电流的负调节剂之一。总之,这项研究首次证实了成年和新生 Nav1.5 同工型在大鼠 DRG 神经元和周围感觉神经元轴突中的表达,但在疼痛模型中它们的表达水平下降。PKC-γ 的上调可能直接或间接下调 SNI 大鼠模型中 Nav1.5 同工型的表达,这可能进一步涉及到神经病理性疼痛的病理过程。