Department of Orthopaedic Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan.
Spine (Phila Pa 1976). 2010 Jun 15;35(14):1346-52. doi: 10.1097/BRS.0b013e3181c099b0.
STUDY DESIGN: We examined the sensory innervation of the coccygeal (Co) 5/6 intervertebral disc in rats using a retrograde neurotracing method and immunohistochemistry. OBJECTIVE: To investigate the properties of the sensory innervation of the rat coccygeal disc. SUMMARY OF BACKGROUND DATA: Developing a rat disease model for degenerative intervertebral disc compression using lumbar discs is technically impractical because of their location. Coccygeal intervertebral discs are more readily accessible and several reports of morphologic evaluation of degenerative coccygeal intervertebral discs using compression devices exist. However, their sensory innervation and properties have not yet been characterized. METHODS: FluoroGold neurotracer was applied to the Co5/6 intervertebral discs of intraperitoneally anesthetized Sprague Dawley rats (n = 10). Subsequently, the discs and the L1-S4 dorsal root ganglions (DRGs) were resected and sectioned. The discs were double-stained for immunoreactivity to the neuronal marker beta-tubulin (Tuj-1) and biotin-labeled isolectinB4 (IB4), a neuropathic pain marker, or Tuj-1 and calcitonin gene-related peptide (CGRP), an inflammatory pain marker. The DRGs were double-stained for IB4-binding and CGRP immunoreactivity (IR). The proportions of IB4-binding or CGRP-IR DRG neurons were assessed by cell counting and compared. RESULTS: The disc immunohistochemistry showed evidence of sensory nerve fibers lying in the outermost layer of the anulus fibrosus. FluoroGold labeled DRG neurons mainly derived from S1 to S3 DRGs, especially S2 and S3. No labeled neurons were observed in the S4 DRG. The histochemistry of the DRGs showed a predominance of CGRP-IR DRG neurons (3.5 +/- 1.7% IB4-binding and 15.4 +/- 5.6% CGRP-IR on average). CONCLUSION: This study showed evidence for nerve fibers in the discs and predominant innervation by CGRP-IR DRG neurons. The neurons innervating the discs mostly derived from S1 to S3 DRGs, especially S2 and S3. These findings may be useful in developing rat models of disease involving degenerative intervertebral disc compression.
研究设计：我们使用逆行神经示踪法和免疫组织化学方法研究了大鼠尾骨（Co）5/6 椎间盘的感觉神经支配。
目的：研究大鼠尾骨椎间盘感觉神经支配的特性。
背景资料概要：由于位置原因，使用腰椎间盘在大鼠身上建立退行性椎间盘压迫的疾病模型在技术上是不切实际的。尾骨椎间盘更容易接近，并且有几篇关于使用压缩装置对退行性尾骨椎间盘进行形态学评估的报告。然而，它们的感觉神经支配和特性尚未得到描述。
方法：将荧光金神经示踪剂应用于腹腔麻醉的 Sprague Dawley 大鼠的 Co5/6 椎间盘（n = 10）。随后，切除椎间盘和 L1-S4 背根神经节（DRG）并进行切片。椎间盘进行神经元标志物β-微管蛋白（Tuj-1）和生物素标记的异硫氰酸荧光素 B4（IB4）双重染色，IB4 是神经病理性疼痛标志物，或 Tuj-1 和降钙素基因相关肽（CGRP），一种炎症性疼痛标志物。DRG 进行 IB4 结合和 CGRP 免疫反应性（IR）双重染色。通过细胞计数评估 IB4 结合或 CGRP-IR DRG 神经元的比例，并进行比较。
结果：椎间盘免疫组织化学显示，感觉神经纤维位于纤维环的最外层。荧光金标记的 DRG 神经元主要来自 S1 到 S3 DRG，特别是 S2 和 S3。在 S4 DRG 中未观察到标记神经元。DRG 的组织化学显示 CGRP-IR DRG 神经元占主导地位（平均 3.5% ± 1.7% IB4 结合和 15.4% ± 5.6% CGRP-IR）。
结论：本研究证明了椎间盘内存在神经纤维，并主要由 CGRP-IR DRG 神经元支配。支配椎间盘的神经元主要来自 S1 到 S3 DRG，特别是 S2 和 S3。这些发现可能对建立涉及退行性椎间盘压迫的疾病的大鼠模型有用。
Spine (Phila Pa 1976). 2010-6-15
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