Glycosylation of Ca(V)3.2 Channels Contributes to the Hyperalgesia in Peripheral Neuropathy of Type 1 Diabetes.
Glycosylation of Ca(V)3.2 Channels Contributes to the Hyperalgesia in Peripheral Neuropathy of Type 1 Diabetes.
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DOI：
10.3389/fncel.2020.605312
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发表时间：
2020
影响因子：
5.3
通讯作者：
Todorovic SM
中科院分区：
文献类型：
--
作者：
Joksimovic SL;Evans JG;McIntire WE;Orestes P;Barrett PQ;Jevtovic-Todorovic V;Todorovic SM
我们以前的研究表明，CaV3.2亚型T-型钙通道(T-通道)的糖基化与2型痛性周围型糖尿病神经病变(PDN)的发生有关。在我们的体外研究中，我们研究了神经氨酸酶(Neu)和PNGase-F(PNG)等去糖基化酶对重组CaV3.2通道的调节机制，以及它们对1型疼痛PDN的行为和生化的影响。在我们的体外研究中，我们使用全细胞电流-电压关系记录来证实去糖基化后CaV3.2的电流密度降低了~2倍。此外，去糖基化导致激活和失活的稳态关系发生显著的去极化转变，而对电流失活和失活恢复的动力学影响很小。用链脲佐菌素(STZ)对成年雌性C57BL/6J野生型(WT)小鼠、成年雌性SD大鼠和CaV3.2基因敲除(KO)小鼠进行体内注射诱导PDN。将Neu或赋形剂(生理盐水)局部注射到右侧后爪或鞘内。我们发现，注射neu，但不注射赋形剂，完全逆转了糖尿病WT大鼠和小鼠的热和机械痛敏。相比之下，neu没有改变CaV3.2KO小鼠的基线温度和机械敏感性，这些小鼠也没有出现疼痛的PDN。最后，我们使用生化方法结合凝胶位移分析直接证明了在健康和糖尿病动物中，背根神经节(DRG)中天然CaV3.2通道的N-末端片段是糖基化的。我们的结果表明，在感觉神经元中，糖基化诱导的CaV3.2通道的改变直接增强了糖尿病的痛觉过敏，糖基化抑制剂可以用于缓解1型糖尿病的疼痛症状。我们期望我们的研究能够更好地理解疼痛的PDN的分子机制，从而促进这种顽固性疾病的新疗法的发现。
Our previous studies implicated glycosylation of the CaV3.2 isoform of T-type Ca2+ channels (T-channels) in the development of Type 2 painful peripheral diabetic neuropathy (PDN). Here we investigated biophysical mechanisms underlying the modulation of recombinant CaV3.2 channel by de-glycosylation enzymes such as neuraminidase (NEU) and PNGase-F (PNG), as well as their behavioral and biochemical effects in painful PDN Type 1. In our in vitro study we used whole-cell recordings of current-voltage relationships to confirm that CaV3.2 current densities were decreased ~2-fold after de-glycosylation. Furthermore, de-glycosylation induced a significant depolarizing shift in the steady-state relationships for activation and inactivation while producing little effects on the kinetics of current deactivation and recovery from inactivation. PDN was induced in vivo by injections of streptozotocin (STZ) in adult female C57Bl/6j wild type (WT) mice, adult female Sprague Dawley rats and CaV3.2 knock-out (KO mice). Either NEU or vehicle (saline) were locally injected into the right hind paws or intrathecally. We found that injections of NEU, but not vehicle, completely reversed thermal and mechanical hyperalgesia in diabetic WT rats and mice. In contrast, NEU did not alter baseline thermal and mechanical sensitivity in the CaV3.2 KO mice which also failed to develop painful PDN. Finally, we used biochemical methods with gel-shift analysis to directly demonstrate that N-terminal fragments of native CaV3.2 channels in the dorsal root ganglia (DRG) are glycosylated in both healthy and diabetic animals. Our results demonstrate that in sensory neurons glycosylation-induced alterations in CaV3.2 channels in vivo directly enhance diabetic hyperalgesia, and that glycosylation inhibitors can be used to ameliorate painful symptoms in Type 1 diabetes. We expect that our studies may lead to a better understanding of the molecular mechanisms underlying painful PDN in an effort to facilitate the discovery of novel treatments for this intractable disease.
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选择性抑制Ca（v）3.2通道逆转周围伤害感受器的过度兴奋，并减轻术后疼痛。
影响因子：
7.3
作者：
Joksimovic SL;Joksimovic SM;Tesic V;García-Caballero A;Feseha S;Zamponi GW;Jevtovic-Todorovic V;Todorovic SM
通讯作者：
Todorovic SM
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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