Retrograde Adenoviral Vector Targeting of Nociresponsive Pontospinal Noradrenergic Neurons in the Rat In Vivo
Retrograde Adenoviral Vector Targeting of Nociresponsive Pontospinal Noradrenergic Neurons in the Rat In Vivo
复制标题
DOI：
10.1002/cne.21879
复制
发表时间：
2009-01-10
影响因子：
2.5
通讯作者：
Pickering, Anthony E.
中科院分区：
文献类型：
--
作者：
Howorth, Patrick W.;Teschemacher, Anja G.;Pickering, Anthony E.
脊髓背角接受来自AS、蓝斑（LC）和A7细胞群的脑桥神经元的密集的含去甲肾上腺素纤维的神经支配。这些脑桥脊髓神经元被认为是构成内源性镇痛系统的组成部分。我们使用了一个腺病毒载体与儿茶酚胺能选择性启动子（AVV-PRS）逆行标记的去甲肾上腺素能神经元投射到腰（L4-L5）背角与增强型绿色荧光蛋白（EGFP）或单体红色荧光蛋白（mRFP）。在将AVV-PRS-EGFP注射到L4-L5的背角后，在A5- 12%、LC-80%和A7-8%中发现了逆行标记的神经元（145 +/- 12，n = 14）。这些神经元对多巴胺β-羟化酶呈免疫阳性，表明它们是儿茶酚胺能的。逆行标记在注射后7天是最佳的，持续超过4周，并且依赖于病毒载体滴度。去甲肾上腺素能投射的脊髓拓扑结构使用EGFP和mRFP表达腺病毒载体进行了检查。脑桥脊髓神经元提供脊髓的双侧神经支配，并且投射到颈和腰区域的神经元的分布几乎没有重叠。用GFP免疫细胞化学观察脑桥脊髓神经元的轴突乔木，以显示向下橄榄核、小脑、丘脑和皮质的投射，但不显示向海马或尾壳核的投射。福尔马林试验诱发这些脑桥脊髓神经元中c-fos表达，表明它们是伤害反应性的（A5- 21%，LC-16%，和A7- 26%，n = 8）。因此，我们已经开发了一种基于病毒载体的策略，选择性地，逆行靶向脑桥脊髓去甲肾上腺素能神经元，可能参与伤害性感受的下行控制。J. Comp.神经元512：141-157，2009. (C)2008 Wiley-Liss，Inc.
The spinal dorsal horn receives a dense innervation of noradrenaline-containing fibers that originate from pontine neurons in the AS, locus coeruleus (LC), and A7 cell groups. These pontospinal neurons are believed to constitute a component of the endogenous analgesic system. We used an adenoviral vector with a catecholaminergic-selective promoter (AVV-PRS) to retrogradely label the noradrenergic neurons projecting to the lumbar (L4-L5) dorsal horn with enhanced green fluorescent protein (EGFP) or monomeric red fluorescent protein (mRFP). Retrogradely labeled neurons (145 +/- 12, n = 14) were found in A5-12%, LC-80% and A7-8% after injection of AVV-PRS-EGFP to the dorsal horn of L4-L5. These neurons were immunopositive for dopamine beta-hydroxylase, indicating that they were catecholaminergic. Retrograde labeling was optimal 7 days after injection, persisted for over 4 weeks, and was dependent on viral vector titer. The spinal topography of the nor-adrenergic projection was examined using EGFP- and mRFP-expressing adenoviral vectors. Pontospinal neurons provide bilateral innervation of the cord and there was little overlap in the distribution of neurons projecting to the cervical and lumbar regions. The axonal arbor of the pontospinal neurons was visualized with GFP immunocytochemistry to show projections to the inferior olive, cerebellum, thalamus, and cortex but not to the hippocampus or caudate putamen. Formalin testing evoked c-fos expression in these pontospinal neurons, suggesting that they were nociresponsive (A5-21%, LC-16%, and A7-26%, n = 8). Thus, we have developed a viral vector-based strategy to selectively, retrogradely target the pontospinal noradrenergic neurons that are likely to be involved in the descending control of nociception. J. Comp. Neurol. 512:141-157, 2009. (C) 2008 Wiley-Liss, Inc.