KATP channel subunits in rat dorsal root ganglia: alterations by painful axotomy.
KATP channel subunits in rat dorsal root ganglia: alterations by painful axotomy.
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DOI：
10.1186/1744-8069-6-6
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发表时间：
2010-01-26
期刊：
影响因子：
3.3
通讯作者：
Sarantopoulos C
中科院分区：
文献类型：
--
作者：
Zoga V;Kawano T;Liang MY;Bienengraeber M;Weihrauch D;McCallum B;Gemes G;Hogan Q;Sarantopoulos C
神经元中的ATP敏感性钾（KATP）通道介导神经保护，它们调节膜兴奋性，并且它们控制神经递质释放。由于DRG神经元KATP电流的损失参与周围神经损伤后疼痛的病理生理学，我们的特点是在大鼠DRG的KATP通道亚单位的分布，并确定其变化通过疼痛轴突切断术，使用RT-PCR，免疫组织化学和电子显微镜。PCR检测到对照DRG神经元中Kir6.1、Kir6.2、SUR 1和SUR 2的转录。除Kir6.1外的所有蛋白表达均通过Western印迹和免疫组织化学证实。这些亚基的免疫染色，通过荧光和共聚焦显微镜确定在质膜和核膜，在胞质溶胶中，沿着外周纤维，并在卫星胶质细胞。Kir6.2与SUR 1亚基共定位。Kir6.2、SUR 1和SUR 2亚基在神经元亚群中被鉴定，通过阳性或阴性NF 200或CGRP染色分类。在切除的补丁记录KATP电流被阻断格列本脲，但预孵育与抗体对SUR 1废除这种阻断作用格列本脲，证实抗体的目标SUR 1蛋白在神经元质膜。在有髓神经纤维中，我们观察到抗SUR 1免疫染色在规则间隔的漏斗状结构。这些结构通过电子显微镜鉴定为由许旺细胞形成的施密特-兰特曼切口（SLI）。在结旁部位，对SUR 1和Kir6.2的免疫染色与抗Caspr共定位。从脊神经结扎致痛的大鼠中切除的DRG表现出与对照组DRG相似的Kir6.2、SUR 1或SUR 2染色，但显示SUR 1免疫荧光NF 200阳性神经元的患病率降低。在DRG和背根近端轴突切断SLI较小，并显示减少SUR 1免疫荧光。我们确定了Kir6.2/SUR 1和Kir6.2/SUR 2 KATP通道在大鼠DRG神经元胞体，周围神经纤维，胶质卫星和雪旺细胞，在正常状态和疼痛神经损伤后。这是第一次报告的KATP通道旁的网站相邻的结结的Ranvier和雪旺细胞的SLI。在疼痛的轴突切断后，KATP通道在大的有髓鞘体和SLI中下调，与对照相比，SLI的尺寸也较小。由于KATP通道在神经元和神经胶质中可能具有不同的功能作用，因此需要进一步研究来探索KATP通道作为神经病理性疼痛和神经退行性变治疗靶点的潜力。
ATP-sensitive potassium (KATP) channels in neurons mediate neuroprotection, they regulate membrane excitability, and they control neurotransmitter release. Because loss of DRG neuronal KATP currents is involved in the pathophysiology of pain after peripheral nerve injury, we characterized the distribution of the KATP channel subunits in rat DRG, and determined their alterations by painful axotomy using RT-PCR, immunohistochemistry and electron microscopy. PCR demonstrated Kir6.1, Kir6.2, SUR1 and SUR2 transcripts in control DRG neurons. Protein expression for all but Kir6.1 was confirmed by Western blots and immunohistochemistry. Immunostaining of these subunits was identified by fluorescent and confocal microscopy in plasmalemmal and nuclear membranes, in the cytosol, along the peripheral fibers, and in satellite glial cells. Kir6.2 co-localized with SUR1 subunits. Kir6.2, SUR1, and SUR2 subunits were identified in neuronal subpopulations, categorized by positive or negative NF200 or CGRP staining. KATP current recorded in excised patches was blocked by glybenclamide, but preincubation with antibody against SUR1 abolished this blocking effect of glybenclamide, confirming that the antibody targets the SUR1 protein in the neuronal plasmalemmal membrane. In the myelinated nerve fibers we observed anti-SUR1 immunostaining in regularly spaced funneled-shaped structures. These structures were identified by electron microscopy as Schmidt-Lanterman incisures (SLI) formed by the Schwann cells. Immunostaining against SUR1 and Kir6.2 colocalized with anti-Caspr at paranodal sites. DRG excised from rats made hyperalgesic by spinal nerve ligation exhibited similar staining against Kir6.2, SUR1 or SUR2 as DRG from controls, but showed decreased prevalence of SUR1 immunofluorescent NF200 positive neurons. In DRG and dorsal roots proximal to axotomy SLI were smaller and showed decreased SUR1 immunofluorescence. We identified Kir6.2/SUR1 and Kir6.2/SUR2 KATP channels in rat DRG neuronal somata, peripheral nerve fibers, and glial satellite and Schwann cells, in both normal state and after painful nerve injury. This is the first report of KATP channels in paranodal sites adjacent to nodes of Ranvier and in the SLI of the Schwann cells. After painful axotomy KATP channels are downregulated in large, myelinated somata and also in SLI, which are also of smaller size compared to controls. Because KATP channels may have diverse functional roles in neurons and glia, further studies are needed to explore the potential of KATP channels as targets of therapies against neuropathic pain and neurodegeneration.
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影响因子：
2.5
作者：
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通讯作者：
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影响因子：
6.2
作者：
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影响因子：
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作者：
Abdulla, FA;Smith, PA
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影响因子：
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作者：
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DOI：
10.1073/pnas.0901815106
发表时间：
2009-05-26
影响因子：
11.1
作者：
Kawano, Takashi;Zoga, Vasiliki;Sarantopoulos, Constantine
通讯作者：
Sarantopoulos, Constantine