We wish to thank Mr Wabbel for preparing the wires and Mr Tietsch for constructing the plugs and cages according to our plans. This work was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 688). Felix Fluri holds a fellowship of the Interdisziplin&#228;re Zentrum f&#252;r Klinische Forschung (IZKF), University Clinics W&#252;rzburg, Germany.

动物实验批准了维尔茨堡大学和法治国家主管部门（下弗兰肯行政区，批准文号:54-2531.01-102 / 13）和执行根据的研究实验性卒中的建议，研究了17和当前的动物研究:的报告体内实验指南（http://www.nc3rs.org.uk/arrive-guidelines）。 1.麻醉<ol><li>检查麻醉剂系统，以确保供应气体（氧气）和异氟烷足量的过程的持续时间。连接的立体定位仪的门牙条鼻锥，把门牙栏上的-3.3毫米。 </li><li>打开供应气体（2L /分钟）。将大鼠成箱和密封的顶部。打开异氟烷蒸发器至3.5％。 </li><li>当老鼠是横卧，切换系统，使麻醉气体流到被固定到所述门齿杆鼻锥。 </li><li>删除RA吨从箱室和刮耳朵和眼睛之间的区域;用棉签浸泡Jodosept PVP，擦拭剃光区域，以消除任何松散的头发。 </li><li>定位在鼻锥（图1）的大鼠和继续用异氟烷2.5％麻醉中的 O 2（1升/分钟）。按捏叉指区域检查麻醉的水平。如果大鼠充分麻醉，防御反射被取消（脚即停药）。 </li><li>手术过程中监测呼吸和刺激反应，并根据需要调整蒸发器。 </li><li>取适量涂抹于眼兽医药膏，以防止干燥，而在麻醉下。监测和反馈控制的加热系统保持体温在37±0.5°C。 </li></ol> 2.手术<ol><li>保持术野消毒，整个手术过程中。一旦surgeon&#39;s手是无菌的并且操作字段是无菌的，只移动carefuLLY记得不要打破不育。这包括也是一个无菌区（ 即，无菌防水窗帘），哪一个可以放下工具。 </li><li>注入0.2 ml的甲哌卡因皮下进入剃区域的中心。甲哌卡因是局部麻醉剂具有至多3小时的作用持续时间。它会进一步麻醉手术区。 </li><li>使用手术刀，使两耳之间中线切口开始并朝向2厘米延伸。确保骨膜（皮肤下有光泽的膜）也被切开。暴露四个夹（ 图2）的头骨。 </li><li>用棉签，轻轻地取下骨膜，直到冠状面和矢状缝暴露;此后，止住血用脱脂棉。 </li><li>确定使用针固定在探头座前囟的坐标，然后标记针用黑色毡尖笔的尖端。使用前/后（AP）中线/外侧（ML）和背腹（DV）驱动螺丝，定位针的尖端直接在囟。 </li><li>参加AP和ML游标刻度读数:减去3.6毫米从AP的阅读和2.5毫米的ML读数电极植入右侧STN，或加入2.5毫米的电极植入左STN。针的尖端下降到颅骨的表面上后，此位置将被标记的毡尖笔的染料。 </li><li>夹紧牙钻到立体定位仪的大探头支架。将牙钻来计算的区域- 即，在头骨的标记点。看，通过显微镜，钻透头骨的孔（直径约1毫米），直到硬脑膜是可见的（头骨为约1毫米厚）。缩回使用显微解剖钳或消毒针头硬脑膜。硬脑膜是艰难的，足以摧毁电极的尖端。 </li><li>上钻孔，在每个正面鳞的牙钻，并且在整型erparietal相对的电极孔鳞。断开立体定位仪的探头支架。不要在颅骨缝钻的静脉血管遵循头骨下缝合。 </li><li>拧接骨螺钉到每一个五杆洞的。避免螺纹螺钉陷得太深。对于不锈钢螺丝（M1.6），2-3圈的螺丝将充分保持螺丝没有把压力对大脑。匝数取决于螺杆的螺距。夹紧探头支架具有（图3）中的显微电极。 </li><li>使用AP，ML和DV的驱动器螺钉，移动探头保持器与电极直到其前端几乎触及前囟。请注意AP，ML和DV游标刻度读数前囟门。当读数制成，提高电极几毫米以防止电极从运动期间刮头骨。为了确定位置的坐标，其中所述电极，必须在插洞口，增加3.6毫米到AP的阅读，并添加（或减）2.5mm至的ML阅读。 </li><li>使用AP和ML驱动螺丝，移动电极所计算的位置。在这一点上，电极尖端应当直接位于在钻孔电极孔。然后，通过查看通过显微镜，降低电极到硬脑膜（图4）的水平。这个水平作为零电平在DV方向。然后，轻轻地通过观察通过显微镜插入电极进入大脑的尖端。 </li><li>连接电极销到记录系统的连接器。放一个法拉第笼（或用铝箔代替它）在大鼠中的立体定位仪（图5）。接地立体定位仪与在正在处理房间的平衡。 </li><li>启动记录系统。如果有的话，还可以使用扬声器advancin过程中获得的排放/单台药膏的声信号克的电极。 </li><li>慢慢推进电极在录制过程中的电活动插入电极进入大脑。在距硬脑膜之间7.5和8.1毫米的深度，在STN的特定电活性通常是可检测的（图6）。在STN神经元的典型活性的特征在于不规则的点火模式和高的发射率（平均频率:40.9±12.9赫兹）18。 </li><li>在记录，减少尽可能麻醉（例如，0.8-1.0％）;低动物麻醉显示更清晰的电子大脑活动。 </li><li>拭远降低电极时被置换在颅骨的表面上的任何血液或脑脊液。 </li><li>混淆少量牙科水泥和应用它周围的电极及其周围的四个使用小刮铲（图7）的五个螺钉。第五螺钉将用于固定插头的接地线。 </li><li>断开记录系统的电极座和连接器的电极销，当牙科用粘固剂固定。 </li><li>拧开，这不是固定的牙科水泥螺丝。将电源插头上的电极针。固定与第五螺钉（图8）插头的接地线。 </li><li>混合了牙科水泥和运用它周围的插头。作为水泥变稠，模具它周围的插头，以形成一个盖。避免牙科水泥可能危害动物和硬化（ 图9A和 B）中删除它们锋利的边缘。 </li><li>清创伤口边缘和带缝线关闭它们的前部和所述盖的后面。消毒伤口边缘。 </li><li>头部插头连接到被固定在一个旋转的导线。从立体定位仪中取出老鼠。 </li><li>每日一次申请的曲马多（12.5毫克/千克，经腹膜内）在干预结束时，然后2-3天。放置老鼠在一个干净的笼THERMAl支持，固定在这个笼子（ 图10）旋转，仔细检查它1小时。 </li><li>不要让动物无人看管，直到它重新获得足够的意识，以保持胸骨斜卧。不返回已经历手术给其他动物的公司，直到完全恢复的动物。 </li></ol> 3.刺激<ol><li>确定使用阻抗计在动物的电阻刺激前。 </li><li>连接旋转的插头用导线和插头在导线与当前输出的另一端和用于刺激器的地线的输出。连接刺激与计算机，以便在节目的刺激器。 </li><li>选择刺激方案的参数;例如，在帕金森氏病中使用的参数是脉冲长度:60微秒;频率:130赫兹。与增加的电流幅值刺激老鼠，直至运动障碍的认可。减少日电子电器强度由强度低于10-20％是引起运动障碍，或直至神经症状消失，所述动物是舒适。单相矩形脉冲在此研究中使用。 </li><li>在完成实验后，安乐死用异氟烷动物:调整isofurane流量或浓度为5％或更大。继续异氟醚曝光，直至呼吸停止后1分钟。 </li></ol>

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The authors declare that they have no competing financial interests.

<html>这项研究提出了一个一步一步的指令集，用于植入单极慢性电极插入老鼠的STN。虽然具有低阻抗钨电极通常用于DBS 18,19，由铂/铱（铂/铱）溶液采用该具有约1兆欧的阻抗的单极电极。铂/铱电极也可用于治疗帕金森氏病，因为它们的有利性质的:它们显示最低侵蚀20，不产生，如果没有高的电荷密度被用于相关组织损伤21。由于本研究的目的是一个长期刺激设置和，以达?...

深部脑刺激（DBS）是一种基于电脉冲通过植入电极，以特定脑结构中，递送一个治疗诸如内部苍白球1，底丘脑核（STN）2 - 4或腹侧中间丘脑5。在过去的二十年中，这种治疗方法已经被确立为一个有力的治疗工具为帕金森氏病1 - 4，张力障碍6和震颤7，并且也用于调节慢性疼痛7，精神障碍（例如，强迫症8，抑郁症9）或难治性癫痫10,11。此外，DBS可能，在未来，成为耐火动脉高血压12或体位性低血压13一种治疗选择。 效果背后的生理机制DBS的仍然知之甚少。在研究啮齿动物的麻醉提供了深入了解，以模仿临床应用DBS 14高频刺激神经 ​​反应。然而，这些研究不仅缺乏的DBS影响行为的佐证，但也导致依赖于刺激参数相当大的变化应用14。 为了研究更简明星展银行在有意识的啮齿类动物行为的影响和作用机制，刺激建立是必要的，满足特定需求。 DBS大多用作长期治疗（例如，帕金森氏病，慢性疼痛）。因此，刺激设置在啮齿类动物的设计应使该单元由一个插头，以及从插头向外部刺激器的导线的电极的;当固定在头骨这个单位应该是轻量级的，但牢不可破。此外，行动自由是必不可少的stimula在大鼠化过长时间。 DBS的目标结构是小的;例如，在大鼠中的STN的长度为1.2毫米和0.8毫米3,15的体积。因此，电极必须被设计为使得细胞核未插入时损伤并靶向需要是精确的。如在啮齿类动物进行最DBS研究已经使用的电极与靶结构基础的里程碑式的立体定向插入时，错误率可以是比较高的，根据Paxinos和Watson 16使用坐标时也是如此。这导致达到统计学上有意义的结果需要的动物的数量较多。 在本研究中的电极注入技术被引入，即通过使用一个microrecording系统，同时推进电极靶向以高精度在STN。此外，刺激系统，提出不仅允许移动性的刺激动物的高度，而且确保连续stimulati上通过的刺激导线的安全固定（这是由一个不锈钢弹簧保护）到大鼠的头部。

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microelectrode guided implantation of electrodes into the subthalamic nucleus of rats for long-term deep brain stimulation

深部脑刺激（DBS）是一种广泛使用的和有效的疗法为几个神经系统疾病，例如特发性帕金森氏病，肌张力障碍或震颤。 DBS是基于递送电刺激中枢神经系统的特定深解剖结构。不过，DBS的效果背后的机制仍然是神秘的。这导致了兴趣调查DBS的在动物模型中的影响，特别是在大鼠。作为星展是一个长期的治疗，研究应集中在星展银行之后发生的数周神经回路的分子遗传学改变。长期DBS在大鼠是具有挑战性的，因为老鼠走动的笼子里，这将导致在保持到位，从动物到刺激的头部领先的电线问题。此外，靶结构刺激大鼠脑内都很小，因此电极不能很容易地放置在所要求的位置上。因此，设置为长效stimula使用铂/铱电极具有大约1兆欧的阻抗大鼠和灰这项研究开发的。这些规范的电极不仅允许足够的刺激，也记录脑深部结构，以确定目标区域的DBS。在我们的设置，用塞子为电线的电极包埋在牙科用粘固剂与固定在颅骨4锚固螺钉。从插头到刺激器的导线是由一个不锈钢弹簧保护。旋转接头被连接到电路，以防止电线的缠结。总体而言，这种刺激设置提供自由流动的高度为大鼠和使头部插头，以及在插头和刺激器之间的导线连接，以保持持久的强度。

植入电极插入用的记录系统大鼠的STN - 作为这里提出 - 是一种有效和精确的程序的DBS该取每只动物约1小时。这个模型是一个相当小的过程:10只进行手术，全部存活的干预。介入后二十四小时，将每只大鼠的状态进行监测，并没有动物根据严重码实现了超过1的3个点。在连续刺激（14天，24小时的天）期间，没有电线脱落，破裂或被咬伤过。没有一个10只失去牙科水泥盖也没有他们得到由设备过程中...

Watch this Scientific Journal Video about Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation at JoVE.com

Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.

微电极引导电极植入到大鼠丘脑底核的长期深部脑刺激

Deep brain stimulation (DBS) is a widely used and effective therapy for several neurologic disorders, such as idiopathic Parkinson&#8217;s disease, ...

microelectrode-guided-implantation-electrodes-into-subthalamic

Research

JoVE Journal

Neuroscience

19.5K 次观看.  University Hospital Wuerzburg. A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.

视频: Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS:使用的Theta突发协议，以探索个人与脆性X综合征与孤独症的可塑性Mechasnism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

科研

神经科学

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic fields. These rapidly alternating fields induce electrical currents within localized, targeted regions of the cortex which are associated with various physiological and functional brain changes.1,2,3
In 2007, O'Reardon et al., utilizing the NeuroStar device, published the results of an industry-sponsored, multisite, randomized, sham-stimulation controlled clinical trial in which 301 patients with major depression, who had previously failed to respond to at least one adequate antidepressant treatment trial, underwent either active or sham TMS over the left dorsolateral prefrontal cortex (DLPFC). The patients, who were medication-free at the time of the study, received TMS five times per week over 4-6 weeks.4
The results demonstrated that a sub-population of patients (those who were relatively less resistant to medication, having failed not more than two good pharmacologic trials) showed a statistically significant improvement on the Montgomery-Asberg Depression Scale (MADRS), the Hamilton Depression Rating Scale (HAMD), and various other outcome measures. In October 2008, supported by these and other similar results5,6,7, Neuronetics obtained the first and only Food and Drug Administration (FDA) approval for the clinical treatment of a specific form of medication-refractory depression using a TMS Therapy device (FDA approval K061053).
In this paper, we will explore the specified FDA approved NeuroStar depression treatment protocol (to be administered only under prescription and by a licensed medical profession in either an in- or outpatient setting).

In this article, we examine the methodology and considerations relevant to the FDA approved depression treatment protocol using the Neuronetics NeuroStar TMS device.

NeuroStar TMS的设备进行美国FDA批准用于治疗抑郁症的议定书"

The Neuronetics NeuroStar Transcranial Magnetic Stimulation (TMS) System is a class II medical device that produces brief duration, pulsed magnetic ...

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form a polished and reinforced thinned skull (PoRTS) window in the mouse skull that spans several millimeters in diameter and is stable for months. The skull is thinned to 10 to 15 &mu;m in thickness with a hand held drill to achieve optical clarity, and is then overlaid with cyanoacrylate glue and a cover glass to: 1) provide rigidity, 2) inhibit bone regrowth and 3) reduce light scattering from irregularities on the bone surface. Since the skull is not breached, any inflammation that could affect the process being studied is greatly reduced. Imaging depths of up to 250 &mu;m below the cortical surface can be achieved using two-photon laser scanning microscopy. This window is well suited to study cerebral blood flow and cellular function in both anesthetized and awake preparations. It further offers the opportunity to manipulate cell activity using optogenetics or to disrupt blood flow in targeted vessels by irradiation of circulating photosensitizers.

We present a method to form an imaging window in the mouse skull that spans millimeters and is stable for months without inflammation of the brain. This method is well suited for longitudinal studies of blood flow, cellular dynamics, and cell/vascular structure using two-photon microscopy.

抛光和钢筋，颅骨变薄窗口，为长期的小鼠脑成像

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard for analyzing patterns of synaptic connectivity, but paired electrophysiological recordings can be both cumbersome and experimentally limiting. The development of optogenetics has introduced an elegant method to stimulate neurons and circuits, both in vitro1 and in vivo2,3. By exploiting cell-type specific promoter activity to drive opsin expression in discrete neuronal populations, one can precisely stimulate genetically defined neuronal subtypes in distinct circuits4-6. Well described methods to stimulate neurons, including electrical stimulation and/or pharmacological manipulations, are often cell-type indiscriminate, invasive, and can damage surrounding tissues. These limitations could alter normal synaptic function and/or circuit behavior. In addition, due to the nature of the manipulation, the current methods are often acute and terminal. Optogenetics affords the ability to stimulate neurons in a relatively innocuous manner, and in genetically targeted neurons. The majority of studies involving in vivo optogenetics currently use a optical fiber guided through an implanted cannula6,7; however, limitations of this method include damaged brain tissue with repeated insertion of an optical fiber, and potential breakage of the fiber inside the cannula. Given the burgeoning field of optogenetics, a more reliable method of chronic stimulation is necessary to facilitate long-term studies with minimal collateral tissue damage. Here we provide our modified protocol as a video article to complement the method effectively and elegantly described in Sparta et al.8 for the fabrication of a fiber optic implant and its permanent fixation onto the cranium of anesthetized mice, as well as the assembly of the fiber optic coupler connecting the implant to a light source. The implant, connected with optical fibers to a solid-state laser, allows for an efficient method to chronically photostimulate functional neuronal circuitry with less tissue damage9 using small, detachable, tethers. Permanent fixation of the fiber optic implants provides consistent, long-term in vivo optogenetic studies of neuronal circuits in awake, behaving mice10 with minimal tissue damage.

The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue.

光纤植入慢性Optogenetic刺激脑组织

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

深部脑刺激与同步功能磁共振成像在鼠害

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Direct Visualization of the Murine Dorsal Cochlear Nucleus for Optogenetic Stimulation of the Auditory Pathway

Investigation into the use of virus-mediated gene transfer to arrest or reverse hearing loss has largely been relegated to the peripheral auditory system. Few studies have examined gene transfer to the central auditory system. The dorsal cochlear nucleus (DCN) of the brainstem, which contains second order neurons of the auditory pathway, is a potential site for gene transfer. In this protocol, a technique for direct and maximal exposure of the murine DCN via a posterior fossa approach is demonstrated. This approach allows for either acute or survival surgery. Following direct visualization of the DCN, a host of experiments are possible, including injection of opsins into the cochlear nucleus and subsequent stimulation by an optical fiber coupled to a blue light laser.&#160;Other neurophysiology experiments, such as electrical stimulation and neural injector tracings are also feasible.&#160;The level of visualization and the duration of stimulation achievable make this approach applicable to a wide range of experiments.

The goal of this protocol is to outline a surgical approach to provide direct access to the dorsal cochlear nucleus in a murine model.

在鼠背耳蜗核的听觉通路的光遗传学刺激直接可视化

Investigation into the use of virus-mediated gene transfer to arrest or reverse hearing loss has largely been relegated to the peripheral auditory system. ...

Analysis of Gene Expression Changes in the Rat Hippocampus After Deep Brain Stimulation of the Anterior Thalamic Nucleus

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective treatment for several movement disorders that have failed to respond to medication. Recent progress in the field of DBS surgery has begun to extend the application of this surgical technique to other conditions as diverse as morbid obesity, depression and obsessive compulsive disorder. Despite these expanding indications, little is known about the underlying physiological mechanisms that facilitate the beneficial effects of DBS surgery. One approach to this question is to perform gene expression analysis in neurons that receive the electrical stimulation. Previous studies have shown that neurogenesis in the rat dentate gyrus is elicited in DBS targeting of the anterior nucleus of the thalamus1. DBS surgery targeting the ATN is used widely for treatment refractory epilepsy. It is thus of much interest for us to explore the transcriptional changes induced by electrically stimulating the ATN. In this manuscript, we describe our methodologies for stereotactically-guided DBS surgery targeting the ATN in adult male Wistar rats. We also discuss the subsequent steps for tissue dissection, RNA isolation, cDNA preparation and quantitative RT-PCR for measuring gene expression changes. This method could be applied and modified for stimulating the basal ganglia and other regions of the brain commonly clinically targeted. The gene expression study described here assumes a candidate target gene approach for discovering molecular players that could be directing the mechanism for DBS.

The mechanism underlying the therapeutic effects of Deep Brain Stimulation (DBS) surgery needs investigation. The methods presented in this manuscript describe an experimental approach to examine the cellular events triggered by DBS by analyzing the gene expression profile of candidate genes that can facilitate neurogenesis post DBS surgery.

基因表达的变化大鼠海马后，前丘脑核脑深部刺激分析

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective ...

Cannula Implantation into the Cisterna Magna of Rodents

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to the skull or the brain parenchyma. In anesthetized rodents, the exposure of the dura mater by blunt dissection of the neck muscles allows the insertion of a cannula into the cisterna magna (CM). The cannula, composed either by a fine beveled needle or borosilicate capillary, is attached via a polyethylene (PE) tube to a syringe. Using a syringe pump, molecules can then be injected at controlled rates directly into the CM, which is continuous with the subarachnoid space. From the subarachnoid space, we can trace CSF fluxes by convective flow into the perivascular space around penetrating arterioles, where solute exchange with the interstitial fluid (ISF) occurs. CMc can be performed for acute injections immediately following the surgery, or for chronic implantation, with later injection in anesthetized or awake, freely moving rodents. Quantitation of tracer distribution in the brain parenchyma can be performed by epifluorescence, 2-photon microscopy, and magnetic resonance imaging (MRI), depending on the physico-chemical properties of the injected molecules. Thus, CMc in conjunction with various imaging techniques offers a powerful tool for assessment of the glymphatic system and CSF dynamics and function. Furthermore, CMc can be utilized as a conduit for fast, brain-wide delivery of signaling molecules and metabolic substrates that could not otherwise cross the blood brain barrier (BBB).

Here we describe a protocol to perform cisterna magna cannulation (CMc), a minimally invasive way to deliver tracers, substrates and signaling molecules into the cerebrospinal fluid (CSF). Combined with different imaging modalities, CMc enables glymphatic system and CSF dynamics assessment, as well as brain-wide delivery of various compounds.

将套管植入啮齿动物的麦格纳池中

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

常见马莫塞的全皮质电皮质阵列的慢性植入

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from parallel fibers (PF) to Purkinje cells (PC) is considered the basis for motor learning, and deficiencies of both LTD and motor learning are observed in various gene-manipulated animals. Common motor learning sets, such as adaptation of the optokinetic reflex (OKR), the vestibular-ocular reflex (VOR), and rotarod test were used for evaluation of motor learning ability. However, results obtained from the GluA2-carboxy terminus modified knock-in mice demonstrated normal adaptation of the VOR and the OKR, despite lacking PF-LTD. In that report, induction of LTD was only attempted using one type of stimulation protocol at room temperature. Thus, conditions to induce cerebellar LTD were explored in the same knock-in mutants using various protocols at near physiological temperature. Finally, we found stimulation protocols, by which LTD could be induced in these gene-manipulated mice. In this study, a set of protocols are proposed to evaluate LTD-induction, which will more accurately allow examination of the causal relationship between LTD and motor learning. In conclusion, experimental conditions are crucial when evaluating LTD in gene-manipulated mice.

In some gene-manipulated animals, using a single protocol may fail to induce LTD in cerebellar Purkinje cells, and there may be a discrepancy between LTD and motor learning. Multiple protocols are necessary to assess LTD-induction in gene-manipulated animals. Standard protocols are shown.

成人脑切片长期抑郁症诱导评估

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...