迷走神经刺激作为一种工具来诱导可塑性相关途径灭绝学习
Summary
迷走神经刺激（VNS）已成为一种工具来促使针对性的突触可塑性的脑修改范围的行为。本协议描述了如何实现VNS方便灭绝的恐惧记忆的巩固。
Abstract
Extinction describes the process of attenuating behavioral responses to neutral stimuli when they no longer provide the reinforcement that has been maintaining the behavior. There is close correspondence between fear and human anxiety, and therefore studies of extinction learning might provide insight into the biological nature of anxiety-related disorders such as post-traumatic stress disorder, and they might help to develop strategies to treat them. Preclinical research aims to aid extinction learning and to induce targeted plasticity in extinction circuits to consolidate the newly formed memory. Vagus nerve stimulation (VNS) is a powerful approach that provides tight temporal and circuit-specific release of neurotransmitters, resulting in modulation of neuronal networks engaged in an ongoing task. VNS enhances memory consolidation in both rats and humans, and pairing VNS with exposure to conditioned cues enhances the consolidation of extinction learning in rats. Here, we provide a detailed protocol for the preparation of custom-made parts and the surgical procedures required for VNS in rats. Using this protocol we show how VNS can facilitate the extinction of conditioned fear responses in an auditory fear conditioning task. In addition, we provide evidence that VNS modulates synaptic plasticity in the pathway between the infralimbic (IL) medial prefrontal cortex and the basolateral complex of the amygdala (BLA), which is involved in the expression and modulation of extinction memory.
Introduction
古典恐惧条件提供了一种广泛使用的动物模型来研究焦虑症的生物学基础。期间恐惧条件，厌恶的刺激（非条件刺激，US，例如 ，一个电击）的输出，结合一个中性刺激，例如铃声和/或上下文（条件刺激; CS）。在恐惧条件，CS和美国之间的关联而形成的。最终，CS的单独呈现一个引起恐惧反应（该条件反应; CR）。在恐惧消退中，CS反复提出在没有美国，造成CR到逐渐减少1。因此，条件性恐惧灭绝是其中可怕的行为反应，以中性刺激衰减，当他们不再预测厌恶成果的活动进程。是不是条件反射灭绝，需要巩固与协会了解到新的竞争回忆。焦虑症的特点是IMPAIRED灭绝2-4。因此，条件性恐惧的动物模型灭绝同时作为抑制性的学习和行为作为治疗人类焦虑症5,6模型的一个重要范例。
因为恐惧和焦虑的人之间的紧密对应关系，它认为这些研究可以提供洞察焦虑有关的疾病如创伤后应激障碍的生物特性，将帮助制定战略对待他们。临床前研究的一个重要目标是帮助灭绝学习和诱导针对性可塑性灭绝电路，以巩固学习灭绝。迷走神经刺激（VNS）是可能被用于提供的脑区和接合在一个正在进行的任务突触紧时间和电路的特定调制微创神经假方法。最近的一系列研究，从迈克尔Kilgard小组在得克萨斯大学达拉斯分校有表明配对VNS与分立感觉或运动的刺激（ 如铃声或杆拉）是促进大脑皮层的可塑性治疗耳鸣7，或中风后8-10克服运动障碍非常有效。此外，非偶然VNS发生在很短的时间窗口后，学习同样促进大脑皮层的可塑性，增强记忆的巩固大鼠和人类中11-13。
考虑在副交感神经通路的迷走神经的作用，这并不奇怪，它可能参与调节记忆和突触可塑性。高度情绪化的事件往往产生于非情感记忆更强的回忆。这可能是由于应激激素对记忆巩固的影响。应激激素肾上腺素Posttraining施用增强记忆巩固在人类和非人类动物，但肾上腺素不穿过血-脑屏障14,15 </suP>。因此，压力引起的肾上腺素释放，必须影响大脑间接地增强记忆的巩固。有力的证据表明，在迷走神经可以是循环肾上腺素和大脑之间的联系。宫下和威廉姆斯16发现，肾上腺素的全身给药提高迷走神经发射，并增加去甲肾上腺素水平的杏仁核17。肾上腺素全身用药不增强记忆的巩固，当β肾上腺素能受体阻断杏仁核18表明迷走神经在，变成情感激发经验转化为长期记忆通路的作用。
因此，VNS配对与培训，以提高支持记忆的巩固和暴露于空调线索在没有加固的增强灭绝学习大鼠19,20巩固脑部变化的潜力。在这里，我们描述了使用的一个VNSSA工具，以促进大脑皮层的可塑性和便利条件性恐惧反应灭绝。
Protocol
在这个协议中描述的所有程序进行，根据美国国立卫生研究院指南实验动物的护理和使用，他们被批准由得克萨斯大学达拉斯分校的机构动物护理和使用委员会。 1.建设VNS袖口创建一个钻井工具被锯掉一个22½摹针的尖底。 运行22个半摹针现在钝端了一个金属文件多次压平。握住针以45度角的文件并运行它多次在该文件同时旋转它。这将导致金属变得更薄和卷起?…
Representative Results
本节中示出了可以通过使用VNS结合灭绝学习，以减少在大鼠条件恐惧反应的表达来获得的结果的例子。对于1和2天（听觉恐惧条件反射），大鼠进行了培训，对听觉恐惧条件任务中footshocks进行搭配的基调。 3（预处理试验）日，音被提出在没有footshocks测量冰点水平，并推断出条件性恐惧反应的收购。第4天收到的（治疗）大鼠组特定消退训练和治疗：4基调演讲中搭配无论是VNS或假刺激或扩展?…
Discussion
我们在这里是，用于曝光的单个会话条件线索19期间促进的条件性恐惧灭绝和调节可塑性在infralimbic皮质和基底外侧杏仁核可能介导消光学20之间的通路的协议。此协议的成功的一个关键步骤是VNS的过程中消退训练正确的交付。因此，应特别注意给予袖带电极的构造和安置周围迷走神经的袖口。在袖带电极的施工过程中，它确保导线的暴露部分是在正确的位置是很重要的。同样地，…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This research was supported by the National Institute of Mental Health MH 086960-01A1 (Christa K. McIntyre).
Materials
| Alcohol | |||
| Atropine | Fisher | A0132-5G | |
| Betadine | Henry Schein | 69066950 | |
| Hydrogen peroxide | CVS | 209478 | |
| Ketamine | Henry Schein | 1129300 | |
| Marcaine | Henry Schein | 6312615 | |
| Mineral Oil | CVS | 152355 | |
| Neosporin | CVS | 629451 | |
| Oxygen | Home Depot | 304179 | |
| Pennicillin | Fisher | PENNA-10MU | |
| Propane | Home Depot | 304182 | |
| Xylazine | Henry Schein | 4019308 | |
| Tools | |||
| Jewelery Torch | Smith Equipment | 23-1001D | |
| Sewing Needle | Walgreens | 441831 | |
| #5 Forceps (2) | Fine Science Tools | 11254-20 | |
| Soldering Iron | Home Depot | 203525863 | |
| AmScope SM-4TX-144A 3.5X-45X Circuit Board Boom Stereo Microscope + 144 LED | AmScope | SM-4TX-144A | |
| Helping Hands | A-M Systems | 726200 | |
| Scalpel Blade Holder | Fine Science Tools | 10003-12 | |
| Metal File | Home Depot | 6601 | |
| Ruler | Home Deopt | 202035324 | |
| Curved Hemostats | Fine Science Tools | 130009-12 | |
| Fine Scissors | Fine Science Tools | 14058-09 | |
| Spatula | Fine Science Tools | ||
| Small Screwdriver | Home Depot | 646507 | |
| Magnetic Fixator Retraction System | Fine Science Tools | 18200-04, 18200-01, 18200-05 | |
| Heating Pad | Walgreens | 30294 | |
| Clippers | Walgreens | 277966 | |
| Sharpie | Staples | 125328 | |
| Ring Forceps | Fine Science Tools | 11103-09 | |
| Custom Micropipette Glass Tools (J shape and Straight) – Borosilicate glass | Sutter Instrument | B150-110-10 | |
| Adson Forceps | Fine Science Tools | 11006-12 | |
| Cuffs | |||
| Tubing | Braintree Scientific Inc | MRE-065 | |
| Platinum Iridium Wire | Medwire | 10IR9/49T | |
| Gold Pins | Mill-Max | 1001-0-15-15-30-27-04-0 | |
| Suture Thread | Henry Schein | 100-5797 | |
| 22 G Needles | Fisher | 14-815-525 | |
| Paper Tape | Fisher | 03-411-602 | |
| Solder | Home Deopt | 327793 | |
| Flux | Home Deopt | 300142 | |
| Scalpel Blade, 10 or 15 | Stoelting | 52173-10 | |
| Silastic Laboratory Tubing .51 mm ID x .94 mm OD | Fisher | 508-002 | |
| Headcaps | |||
| Connector Pieces (male) | Omnetics Connector Corporation | A25001-004 | |
| Headcap pieces (female) | Omnetics Connector Corporation | A24001-004 | |
| Teets Dental Acrylic, Liquid and Powder | A-M Systems | 525000, 526000 | |
| 26 Gauge Solid Copper Wire | Staples | 1016882 | |
| Surgery | |||
| Bone Screws | Stoelting+CB33:C61 | 51457 | |
| Scalpel Blades, 10 or 15 | Stoelting | 52173-10 | |
| 1 ml syringes | Fisher | 14-826-261 | |
| 22 G Needles | Fisher | 14-815-525 | |
| 27 G Needles | Fisher | 14-826-48 | |
| 2" x 2" Gauze | Fisher | 22-362-178 | |
| Swabs | Fisher | 19-120-472 | |
| Puppy Pads | PetCo | 1310747 | |
| Kim Wipes | Fisher | 06-666-A | |
| Chamber and Behavioral Setting | |||
| Husky Metal Front Base Cabinet (30WX19DX34H) | Home Depot | 100607961 | |
| Quiet Barrier HD Soundproofing Material (Sheet) (PSA) | soundproofcow.com | 10203041 | |
| Convoluted Acoustic Foam Panel | soundproofcow.com | 10432400 | |
| Isolated Pulse Stimulator Model 2100 | A-M Systems | 720000 | |
| Digital Camera – Logitech Webcam C210 | Logitech | B003LVZO88 | |
| MatLab | Mathworks.com | ||
| Sinometer 10MHz Single Channel Oscilloscope | Sinometer | CQ5010C | |
| OxyLED T-01 DIY Stick-on Anywhere 4-LED Touch Tap Light | OXYLED | B00GD8OKY0 | |
| 5k ohm potentiomter | Alpha Electronics | B00CTWDHIO | |
| Extech 407730 40-to-130-Decibel Digital Sound Level Meter | Extech Instruments | B000EWY67W | |
| DSCK-C Dual Output, scrambled shocker | Kinder Scientific Co | ||
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Cite This Article
Childs, J. E., Alvarez-Dieppa, A. C., McIntyre, C. K., Kroener, S. Vagus Nerve Stimulation as a Tool to Induce Plasticity in Pathways Relevant for Extinction Learning. J. Vis. Exp. (102), e53032, doi:10.3791/53032 (2015).