Changes in GAD- and GABA- immunoreactivity in the spinal dorsal horn after peripheral nerve injury and promotion of recovery by lumbar transplant of immortalized serotonergic precursors
Changes in GAD- and GABA- immunoreactivity in the spinal dorsal horn after peripheral nerve injury and promotion of recovery by lumbar transplant of immortalized serotonergic precursors
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DOI：
10.1016/s0891-0618(98)00062-3
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发表时间：
1998-12-01
影响因子：
2.8
通讯作者：
Montanez, K
中科院分区：
文献类型：
--
作者：
Eaton, MJ;Plunkett, JA;Montanez, K
我们利用 RN46A 细胞（一种源自 E13 脑干中缝的永生化神经元细胞系）作为生物工程血清素能细胞移植的模型。 RN46A 细胞需要脑源性神经营养因子 (BDNF) 来提高体外和体内的存活率和血清素 (5HT) 合成。将大鼠BDNF基因转染RN46A细胞，亚克隆46A-B14细胞系。这些细胞移植到腰脊髓蛛网膜下腔后存活时间超过 7 周，并合成 5HT 和 BDNF。坐骨神经的慢性压缩损伤（CCI）被用来诱导大鼠受影响后爪的慢性神经性疼痛。 CCI后1周移植的46A-B14细胞减轻了慢性神经性疼痛，而5HT阴性且不含BDNF基因的46A-V1对照细胞的移植对热和触觉伤害感受的诱导没有影响。 CCI 后 3 天和 1-8 周，当对腰脊髓中含有神经递质 γ-氨基丁酸 (GABA) 及其合成酶谷氨酸脱羧酶 (GAD) 的背角内源性细胞进行免疫组织化学定量时，双侧 GABA 和 CAD 免疫反应性 (ir) 细胞的数量减少至 CCI 后 3 天即出现神经损伤。 CCI后1周，双侧GABA-ir细胞数量继续显着下降，到神经损伤后8周，神经损伤对侧的GABA-ir细胞数量恢复到接近正常数量。 CCI后1周，神经损伤两侧的GAD-ir细胞数量开始增加，并在神经损伤后8周继续显着增加，超过正常值。 CCI加细胞移植后2周和8周检查时，46A-B14细胞移植在移植后1周逆转了GAD-ir细胞数量的增加和GABA-ir细胞的减少，而CCI后46A-V1对照细胞移植对GAD-ir或GABA-ir细胞数量的变化没有影响。总的来说，这些数据表明，与移植相关的 5HT 水平的改变，或许还有 BDNF 的分泌，可以改善慢性疼痛，并逆转背角内源性疼痛机制的诱导和维持。这种诱导机制可能依赖于 CCI 启动的 GAD 调节和 GABA 合成的改变。 (C) 1998 Elsevier Science B.V. 保留权利。
We have utilized RN46A cells, an immortalized neuronal cell line derived from E13 brainstem raphe, as a model for transplant of bioengineered serotonergic cells. RN46A cells require brain-derived neurotrophic factor (BDNF) for increased survival and serotonin (5HT) synthesis in vitro and in vivo. RN46A cells were transfected with the rat BDNF gene, and the 46A-B14 cell line was subcloned. These cells survive longer than 7 weeks after transplantation into the subarachnoid space of the lumbar spinal cord and synthesize 5HT and BDNF. Chronic constriction injury (CCI) of the sciatic nerve was used to induce chronic neuropathic pain in the affected hindpaw in rats. Transplants of 46A-B14 cells placed 1 week after CCI alleviated chronic neuropathic pain, while transplants of 46A-V1 control cells, negative for 5HT and without the BDNF gene, had no effect on the induction of thermal and tactile nociception. When endogenous cells of the dorsal horn which contain the neurotransmitter gamma-aminobutyric acid (GABA) and its synthetic enzyme glutamate decarboxylase (GAD) were immunohistochemically quantified in the lumbar spinal cord 3 days and 1-8 weeks after CCI, the number of GABA- and CAD-immunoreactive (ir) cells decreased bilateral to the nerve injury as soon as 3 days after CCI. At 1 week after CCI, the number of GABA-ir cells continued to significantly decline bilaterally, returning to near normal numbers on the side contralateral to the nerve injury by 8 weeks after the nerve injury. The number of GAD-ir cells began to increase bilaterally to the nerve injury at I week after CCI and continued to significantly increase in numbers over normal values by 8 weeks after the nerve injury. When examined 2 and 8 weeks after CCI plus cell transplants, the transplants of 46A-B14 cells reversed the increase in GAD-ir cell numbers and the decrease in GABA-ir cells by 1 week after transplantation, while 46A-V1 control cell transplants after CCI had no effect on the changes in numbers of GAD-ir or GABA-ir cells. Collectively, these data suggest that altered 5HT levels, and perhaps BDNF secretion, related to the transplants ameliorate chronic pain and reverse the induction and maintenance of an endogenous pain mechanism in the dorsal horn. This induction mechanism is likely dependent on altered GAD regulation and GABA synthesis, initiated by CCI. (C) 1998 Elsevier Science B.V. an rights reserved.