Selective optogenetic activation of NaV1.7-expressing afferents in NaV1.7-ChR2 mice induces nocifensive behavior without affecting responses to mechanical and thermal stimuli.
Selective optogenetic activation of NaV1.7-expressing afferents in NaV1.7-ChR2 mice induces nocifensive behavior without affecting responses to mechanical and thermal stimuli.
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DOI：
10.1371/journal.pone.0275751
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
在脊髓背根神经节小神经元和大神经元中，电压门控钠通道亚型NaV1.7、NaV1.8和NaV1.9的表达具有特征性的局域性，可能在疼痛传递和顽固性疼痛的发生中发挥不同的作用。每种特定亚型在体内的选择性刺激可能阐明其在疼痛中的作用。到目前为止，这在现有技术下还很困难。然而，最近发展起来的光遗传学技术已经能够在体内选择性地激活或抑制特定的神经循环。此外，光遗传学甚至被用来选择性地激发表达nav1.8的背根神经节神经元来诱导有害行为。近年来，CRISPR/Cas9等基因修饰技术不断进步，利用该技术可以很容易地产生各种敲入小鼠。我们的目的是研究选择性光遗传激活表达nav1.7的传入神经对小鼠行为的影响。我们利用CRISPR/ cas9介导的同源重组技术，产生了双双子NaV1.7 - iCre敲入小鼠，在内源性NaV1.7基因启动子下表达iCre重组酶，而不破坏NaV1.7。将cre驱动小鼠与通道视紫红素-2 (ChR2) cre报告基因Ai32小鼠杂交，获得NaV1.7iCre/+；Ai32 / +, NaV1.7iCre / iCre;Ai32 / + NaV1.7iCre / +;Ai32/Ai32和NaV1.7iCre/iCre；Ai32 / Ai32老鼠。与野生型小鼠的行为相比，上述基因型小鼠在机械和热刺激相关的行为上没有观察到差异，这表明内源性NaV1.7基因不受iCre靶向插入的影响。蓝光照射后爪后，所有基因型小鼠均以光功率依赖的方式产生足退缩。蓝光房间中爪退缩和厌恶行为的阈值和发生率与ChR2表达水平有关；NaV1.7iCre/iCre反应最强；Ai32 / Ai32老鼠。因此，我们建立了一种非侵入性疼痛模型，在自由运动的转基因NaV1.7-ChR2小鼠中，外周伤害感受器被光激活。
In small and large spinal dorsal root ganglion neurons, subtypes of voltage-gated sodium channels, such as NaV1.7, NaV1.8, and NaV1.9 are expressed with characteristically localized and may play different roles in pain transmission and intractable pain development. Selective stimulation of each specific subtype in vivo may elucidate its role of each subtype in pain. So far, this has been difficult with current technology. However, Optogenetics, a recently developed technique, has enabled selective activation or inhibition of specific neural circulation in vivo. Moreover, optogenetics had even been used to selectively excite NaV1.8-expressing dorsal root ganglion neurons to induce nocifensive behavior. In recent years, genetic modification technologies such as CRISPR/Cas9 have advanced, and various knock-in mice can be easily generated using such technology. We aimed to investigate the effects of selective optogenetic activation of NaV1.7-expressing afferents on mouse behavior. We used CRISPR/Cas9-mediated homologous recombination to generate bicistronic NaV1.7–iCre knock-in mice, which express iCre recombinase under the endogenous NaV1.7 gene promoter without disrupting NaV1.7. The Cre-driver mice were crossed with channelrhodopsin-2 (ChR2) Cre-reporter Ai32 mice to obtain NaV1.7iCre/+;Ai32/+, NaV1.7iCre/iCre;Ai32/+, NaV1.7iCre/+;Ai32/Ai32, and NaV1.7iCre/iCre;Ai32/Ai32 mice. Compared with wild–type mice behavior, no differences were observed in the behaviors associated with mechanical and thermal stimuli exhibited by mice of the aforementioned genotypes, indicating that the endogenous NaV1.7 gene was not affected by the targeted insertion of iCre. Blue light irradiation to the hind paw induced paw withdrawal by mice of all genotypes in a light power-dependent manner. The threshold and incidence of paw withdrawal and aversive behavior in a blue-lit room were dependent on ChR2 expression level; the strongest response was observed in NaV1.7iCre/iCre;Ai32/Ai32 mice. Thus, we developed a non-invasive pain model in which peripheral nociceptors were optically activated in free-moving transgenic NaV1.7–ChR2 mice.
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影响因子：
4.6
作者：
Maimon BE;Diaz M;Revol ECM;Schneider AM;Leaker B;Varela CE;Srinivasan S;Weber MB;Herr HM
通讯作者：
Herr HM
影响因子：
25
作者：
通讯作者：
--
影响因子：
7.4
作者：
Samineni VK;Yoon J;Crawford KE;Jeong YR;McKenzie KC;Shin G;Xie Z;Sundaram SS;Li Y;Yang MY;Kim J;Wu D;Xue Y;Feng X;Huang Y;Mickle AD;Banks A;Ha JS;Golden JP;Rogers JA;Gereau RW 4th
通讯作者：
Gereau RW 4th
影响因子：
46.9
作者：
通讯作者：
--
DOI：
10.1523/jneurosci.3248-09.2009
发表时间：
2009-10-21
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Wang H;Zylka MJ
通讯作者：
Zylka MJ