作者没有确认。

根据机构指南, 动物被安置在南丹麦大学动物保育设施中。所有动物实验都是按照国家卫生研究院的指南进行的, 用于实验室动物的护理和使用。这是一个急性的, 非生存的手术程序。
1. 导管、电极及手术器械的制备
<ol><li>使用以下显微外科器械: 手术剪刀, 角度微剪刀, 纸巾钳, 一对 #7 和 #5 弯曲的微型钳, 微型针持有人, 和拉钩。 注意:由于这是一个急性的程序, 这些仪器不需要消毒。使用后, 清洁和擦拭的提示与70% 乙醇。</li>
	<li>将油管浸泡在70% 乙醇中, 然后在使用前用 100 U/毫升肝素冲洗0.9% 氯化钠。保持油管填充, 以避免引入气泡进入系统。</li>
	<li>将20-30 厘米长的聚乙烯 (PE)-50 油管切割成导管进行 ICP 测量。确保油管尽可能短, 以减少阻尼压力。</li>
	<li>弯曲一个无菌的24G 针的一侧到一边, 直到针打破中间。将该片与斜角连接到 PE 导管的远端, 以便插入到小弟弟小腿中。将另一半插入轮毂以连接到压力传感器。用血气盐水 (100 U/毫升) 填充系统。</li>
	<li>为了使双极聚四氟乙烯涂层电极, 切割两个125µm 银线, 以同等长度。使用一块胶带将电线连接到桌子的边缘, 并将它们串在一起。随后, 将电极连接到黑色板上。</li>
	<li>在聚四氟乙烯上做一个小切口, 用 #5 微钳将4-5 毫米长的聚四氟乙烯涂层从电极的尖端上剥开。用手术刀剪下窍门, 以达到均匀长度, 并通过弯曲刀刃钝边缘周围的两端来创建钩子。</li>
	<li>将电极贴在黑色板的边缘上, 用钩指向向上。十年代将硅胶混合在塑料板上, 将胶泡在电极周围1-2 毫米的钩子上包裹起来。</li>
	<li>在使用前允许它干燥大约5分钟 (图 1)。从另一端的较长的部分剥离聚四氟乙烯, 以允许与刺激器连接。 注意:随着在远端的钩重新做, 电极可以重复使用多次。</li>
</ol>2. 动物的制备
<ol><li>麻醉后, 将腹部、颈部和会阴下半部剃去。用70% 酒精擦洗动物, 接着是聚维酮碘三倍。把老鼠放在一个仰卧位置的加热手术垫上。应用兽医眼膏, 将麻醉转到鼻锥, 2.5% 异氟醚和0.8 升/分钟氧作为载体。 注意:调整异氟醚和氧气的水平, 以达到可接受的麻醉水平。</li>
</ol>3. 术前准备
<ol><li>在操作显微镜下完成整个手术过程: 从3.15X 到20X 的放大倍数是足够的。在整个手术中使用手套和保持一个干净的环境。把老鼠放在褶皱上。</li>
</ol>4. ICP 测量用 Ischiocavernosus 肌清扫术
<ol><li>使用手术刀, 直剪刀和 #7 弯曲的微型钳, 使1厘米垂直皮肤切口5毫米侧向中线开始在基部的小弟弟和向下延伸 (图 2A)。使用 Q 尖和仔细分开的筋膜侧向阴囊 (图 2B)。解剖筋膜后, 附上拉钩和触及与棉花倾斜交换找到坐骨股骨 (图 2C)。</li>
	<li>解剖通过脂肪组织内侧的这一点, 直到 ischiocavernosus 肌肉可视化 (图 3A)。使用一对 #7 弯曲的微型钳和纵向分开的肌肉。膜将显示为明亮的白色结构 (图 3B)。使用微钳和微剪刀, 充分暴露膜, 以查看其课程 (图 3C)。</li>
	<li>校准系统设置后 , 通过会的皮肤插入油管 , 确保它与小弟弟小腿平行 (图 4) 。保持线到位, 使切口湿润生理盐水。</li>
</ol>5. 用于刺激的 CN 解剖
<ol><li>做一个2厘米低, 中线腹部切口通过皮肤使用, 首先, 手术刀, 然后一对直剪刀和微钳。创建一个匹配切口通过筋膜沿 linea 和底层肌肉组织暴露膀胱和前列腺。</li>
	<li>使用拉钩达到良好的曝光率。使用棉尖拭子分离前列腺从脂肪组织, 以获得清晰可视化的主要骨盆神经节 (加仑) 和 CN, 运行在侧方面的前列腺 (图 5)。</li>
	<li>在对加仑和 CN 的可视化后, 小心地切开上面2-5 毫米神经的筋膜, 用角度微剪刀 (图 6a)。使用 #5 微型钳, 传播神经的每一侧和它下面的组织, 以释放4毫米长的部分 (图 6b), 并在神经下滑动9-0 缝合 (图 6c)。</li>
	<li>在缝合 (图 7a) 的帮助下轻微地提升神经, 以便在神经周围放置双极电极的钩子 (图 7b)。让助手将双组分硅胶与胰岛素针的尖端混合在一起5秒. 干神经, 将胶水涂在钩子和神经周围的区域 (图 7c, d)。在电极上稍微拉上一点, 使其神经升高大约1分钟, 让胶水干。</li>
	<li>除去拉钩, 除了右侧的拉钩, 以避免任何拉或扭的电极。用生理盐水将裸露的器官湿润, 在切口上浸泡在生理盐水中。</li>
</ol>6. ICP 测量用海绵体插管法
<ol><li>用拉钩恢复膜的可视化。确保不要将拉钩附着在 ischiocavernosus 肌肉上, 因为它会扭曲小腿。</li>
	<li>将针和血气盐水冲洗油管, 然后再将其引入膜。保持膜拉伸, 使用 #7 弯曲的微型钳一只手 (非显性), 持有的膜和其余的上覆肌肉远端到插入点。用直的微钳握住针在另一主导的手, 并确保介绍它与海绵体的过程平行 (图 8)。</li>
	<li>将针5-8 毫米推入海绵体。冲洗油管并按下小腿以测试线路 (图 9)。确保没有泄漏。用胶带将油管固定在桌子上, 以防意外拉线。删除拉钩。</li>
</ol>7. 对 CN 的刺激
<ol><li>与录音程序 (例如, 穗 2) 运行, 连续记录海绵和平均动脉压力。</li>
	<li>在刺激器上设置以下参数 (例如, SD9 草仪器, 请参见材料表), 用于 CN 刺激: 电流为1.5 毫安, 频率为16赫兹, 电压为3伏, 脉宽为5毫秒. 应用五十年代刺激以极小值 1 min在刺激之间休息。 注意:第一次刺激通常会导致响应减少 (图 10)。</li>
</ol>

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作者没有什么可透露的。

本研究的主要目的是描述一个简化的手术技术的小弟弟小腿插管的 ICP 记录和分离的 CN 为刺激。我们介绍了海绵体解剖的修改, 以简化手术, 并提供可重现性的记录, 增加的 ICP 与 CN 刺激。与1厘米垂直皮肤切口, 侧向基部的小弟弟, 用可触摸的坐骨股骨作为指导, 我们取得了良好的暴露 ischiocavernosus 肌肉和膜。此过程比文献中描述的更快 (低于15分钟), 导致最小组织中断2,3,4。
目前使用的 cn 刺激技术包括在每个刺激应用10之前, cn 的挂钩、提升和烘干。这种技术不能保证每种刺激条件都保持不变。此外, 解除神经需要使用机器人和反复伸展和释放的神经, 这可能导致 neuropraxia。与目前的 CN 刺激技术相比, 使用生物相容性的硅胶分离神经电极复合体, 可减少神经的频率, 伸展到两个情况;一旦将两极电极放在神经周围, 一旦用胶水隔离神经和电极。随后, 可以在不需要神经操作的情况下进行多种 neurostimulations。
将针头插入海绵体内进行压力记录是一个关键步骤。这种技术的新用户在开始实验之前应该练习这一部分。当将针头部署到小腿时, 小腿的拉伸和针与它的航向平行是至关重要的。另一个关键步骤是神经处理过程中的解剖从周围的组织和放置电极下方。虽然解剖本身不是一个困难的任务, 任何伸展或粉碎的神经可能会造成损害。我们对这一步骤的修改使得需要机器人不必要, 并简化了神经处理。使用生物相容性的硅胶水隔离神经和提供稳定和可靠的接触电极和神经, 减少必要的操作。硅胶胶也可用于其他动物模型中, 其中应用了体内神经刺激。
许多研究中列出的刺激参数不同于14-20 赫兹和 1.5-12 V11,13,14。这项技术表明, 刺激使用 1.5 mA, 16 赫兹, 3 V, 和 5 ms 脉冲产生了充分的反应。随着刺激参数的增加, ICP 不会增加 (图 18)。这表明, 刺激使用任何参数, 这是高于阈值触发动脉 (海绵状动脉), 动脉和正弦平滑肌肉松弛, 足以触发反射, 并将导致一个完整的生理响应。随后增加的频率, 振幅, 或脉宽不会导致更强的生理反应, 它可能耗尽神经递质甚至导致神经损伤。通过上述参数, 我们能够实现一个可重现的响应, 其休息期短于三十年代, 连续40次。
麻醉深度明显影响生理反应。随着吸入麻醉, 它可以很好地控制, 但峰值反应约25% 低, 与芬太尼/咪达唑仑麻醉治疗注射。我们观察到, 异氟醚的剂量可以维持在水平确定的最小异氟醚浓度, 以消除脚趾捏反应。大多数调查人员, 使用戊巴比妥钠, 芬太尼/咪唑安定, 氯胺酮/甲苯噻嗪, 或氯胺酮/咪唑安定的注射麻醉,15。
所有发表的文章都没有提到使用控制的氧合。研究结果表明, 这对 ICP 产生了重大影响。因此, 重要的是, 无论使用何种麻醉方式, 动物都能通过鼻锥得到氧气。
使用大鼠是有利的, 由于其大小和韧性。在以前发表的研究和证实在这项研究中, 大大鼠已经被证明对 CN 刺激在6-12 周大的范围内有一个完整的反应, 重200-550 克. 使用小鼠更具挑战性, 但得益于转基因技术的可用性12。

体内对实验动物勃起功能的研究始于 1863年, 埃克哈德1的开创性实验工作。刺激的骨盆神经被用来诱发更高的 ICP 的狗。在整个 20th世纪, 类似的实验性协议被用于大型动物, 如狗, 猴子, 猫和兔子。在 1989年2中, 由昆兰et 等) 首次开发出大鼠勃起功能的评价。此后, 该方法已被其他几个组34修改和更新。今天, 大鼠是最广泛使用的动物模型, 研究勃起功能障碍的病理学和评估新的治疗方案。该程序的主要步骤包括, 记录全身血压使用颈动脉线, 插管的小弟弟小腿, 以测量 icp, 并刺激 CN 诱导增加的 icp。虽然有几个研究人员对模型进行了改进, 但其重现性仍然是一个问题, 不同的实验室也报告了可变的结果。有几个陷阱仍然存在。
上一篇文章5,6,7,8,9,10描述使用全小弟弟暴露与脱的生殖器海绵体插管。这不是一个最佳的方法, 因为操纵和破坏性解剖导致结构损伤, 这是必不可少的勃起功能。CN 的解剖已被很好地描述为10,11, 但是由于多种因素可能会影响实验结果, 神经的刺激不是最佳的。CN 刺激技术包括通过拉住神经周围的双极钩电极来解除周围组织的神经, 并在每次刺激前干燥神经。这可能导致各种程度的神经损伤和电流泄漏, 从而导致通过刺激替代途径例如、盆底肌肉、膀胱和胃肠道而减少反应或假增加 ICP道12。所有这些因素都限制了重现性。
在我们的研究中, 我们观察到, 麻醉的深度和类型都对 ICP 产生了深远的影响。使用的麻醉剂是戊巴比妥钠, 氯胺酮/甲苯噻嗪或氯胺酮/咪达唑仑注射液, 或异氟醚/氧吸入。
在这里, 我们描述了一个简化的手术方法, 并提供数据支持标准化的实验协议。

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			<td height="21" style="height:21px;width:257px;">Dumont #7 forceps</td>
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			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Dumont #5 forceps</td>
			<td style="width:199px;">Fine Science Tool</td>
			<td style="width:119px;">11273-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">ToughCut Mayo scissor</td>
			<td style="width:199px;">Fine Science Tool</td>
			<td style="width:119px;">14110-15</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Miniature Vannas Spring scissor</td>
			<td style="width:199px;">Fine Science Tool</td>
			<td style="width:119px;">15006-09</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Ultra Fine Hemostat</td>
			<td style="width:199px;">Fine Science Tool</td>
			<td style="width:119px;">13020-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Crile Hemostat</td>
			<td style="width:199px;">Fine Science Tool</td>
			<td style="width:119px;">13004-14</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Kwik-Sil Adhesive</td>
			<td style="width:199px;">World Precision Instruments</td>
			<td style="width:119px;">KWIK-SIL</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Teflon coated silver wire 0.125 mm</td>
			<td style="width:199px;">World Precision Instruments</td>
			<td style="width:119px;">AGT0510</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Elastic wire retractors</td>
			<td style="width:199px;">Custom made</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Scalpel blade</td>
			<td style="width:199px;">Fine Science Tool</td>
			<td style="width:119px;">10023-00</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">PE-50 tubing</td>
			<td style="width:199px;">Warner Instruments</td>
			<td style="width:119px;">64-0753</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">23 G Needle</td>
			<td style="width:199px;">Kruuse</td>
			<td style="width:119px;">121272</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">SD-9 Square Pulse Stimulator</td>
			<td style="width:199px;">Somatco</td>
			<td style="width:119px;">1077/183</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Blood pressure transducer and cable</td>
			<td style="width:199px;">World Precision Instruments</td>
			<td>BLPR2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Raucotupf Cotton-tipped Applicators</td>
			<td style="width:199px;">Lohmann-Raucher</td>
			<td style="width:119px;">11966</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Pro-ophta Ocular Sticks</td>
			<td style="width:199px;">Lohmann-Raucher</td>
			<td style="width:119px;">16515</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">NaCl 0,9 % 100 mL</td>
			<td style="width:199px;">Local pharmacy</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Heparin</td>
			<td style="width:199px;">Local pharmacy</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">25 mL Syringe</td>
			<td style="width:199px;">Odense University Hospital</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Vet eye ointment - Viscotears</td>
			<td style="width:199px;">Local pharmacy</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">silver wires&#160;</td>
			<td colspan="2">Science Products GmbH, Heidelberg, Germany</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:257px;">Silicon Glue&#160;</td>
			<td colspan="3">Kwik-Sil, World Precision Instruments, Sarasota, FL, USA</td>
		</tr>
	</tbody>
</table>

cavernous nerve stimulation and recording of intracavernous pressure in a rat

海绵状神经 (CN) 的刺激和海绵压力的测量已被广泛用于测试和评估勃起功能障碍的治疗方法。然而, 所使用的方法在实验室之间有所不同, 而且仍然存在陷阱。这项研究的目的是描述一种外科技术, 将提供一个可靠的和可重复的模式。通过暴露 ischiocavernosus 肌肉在其插入点上的坐骨股骨, 小弟弟小腿可能是空心的最小解剖和伤害的结构涉及勃起功能。在不需要提升和干燥的情况下, 对 CN 进行反复刺激, 通过使用125µm 双极银电极和生物相容的硅胶来分离电极-神经复合体。该方法通过减少伸展和干燥神经, 提供完全的神经隔离, 消除电漏, 防止刺激的替代途径, 防止 neuropraxia。

使用本协议的建议刺激设置, 在吸入麻醉下, 异氟醚2.0% 氧0.8 升/分, 应产生的结果, 如图 11和图12, 其中有多个回到背面刺激之间的75-80 毫米汞.图 13显示与20分钟的激发相同的稳定响应, 其响应稳定在73-77 毫米汞. 通过冲洗管道和敲击小腿 (图 9) 来测试 ICP 测量的线路。回复基线的快速反应是一条很好的线的标志。如果膜的完整性受损, 测试将导致在冲洗和攻丝后的峰值压力降低, 响应速度减慢, 并在激发时响应减弱 (图 14)。在刺激过程中冲洗和流血时, 也会有血气氯化钠的渗漏。
麻醉的类型和水平以及氧气的使用对 ICP 产生了重大影响。图 15演示了不同级别的异氟醚对 ICP 的影响, 其响应减少, 且稳定性较差。与异氟醚在 2%, 在78毫米汞多刺激的 ICP 测量有一个稳定的反应. 然而, 将异氟醚的浓度增加到 3.5%, 导致在多次, 随后的刺激下快速50% 减少到34毫米汞。在将异氟醚从2.0% 转到3.0% 时观察到了同样的效果, 观察到了19% 的反应减少, 从2.5% 到 5%, 在反应的70% 中, 出现了更迅速的减少。在所有刺激中血压保持稳定。在使用异氟醚/氧麻醉的大鼠在手术和最初的刺激, 然后收到25% 的建议剂量芬太尼/咪达唑仑 (而异氟醚停止), 有类似的稳定反应, 但它与异氟醚相比, 芬太尼/咪唑仑麻醉期间增加 25% (图 16)。
通过鼻锥的氧气管理增加了血液中的氧气饱和度从61-75% 到99-100% 在大约二十年代。当氧合停止时, 同样的减少在大约1分钟内被看见在刺激期间血压是稳定的, 但通过鼻子锥体的氧气管理 (0.8 升/分钟) 对最大 ICP 测量有很大影响, 减少它由35-45% 在后向后退刺激 (图 17) 中。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56807/56807fig1.jpg"> 图 1.双极聚四氟乙烯涂层银电极.(A) 在涂层和未涂布电极之间的过渡区内粘附气泡。(B) 电极的远端2厘米紧 braded。(C) 平行未涂覆的钩1毫米分开。<a href="/files/ftp_upload/56807/56807fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56807/56807fig2.jpg"> 图 2.海绵体的解剖 - 地标.(A) 1 厘米垂直皮肤切口, 向下开始2毫米侧向从基部的鸡巴。(B) 用棉尖拭子分离阴囊的侧筋膜。(C) 拉钩放置后的操作字段视图 (pm: 新疆杨肌肉, 它: 小腿的插入点到坐骨股骨)。<a href="/files/ftp_upload/56807/56807fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56807/56807fig3.jpg"> 图 3.暴露膜.(A) ischiocavernosus 肌肉 (箭头)。(B) 膜 (箭头)。(C) 显示海绵体的过程的低功耗放大倍数。<a href="/files/ftp_upload/56807/56807fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56807/56807fig4.jpg"> 图 4.海绵状压力记录线.针通过皮肤在会平行运行与海绵体 ( 箭头 ) 。<a href="/files/ftp_upload/56807/56807fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56807/56807fig5.jpg"> 图 5.在前列腺的侧方面, 由箭头和 CN 垂直运行的加仑暴露.主要骨盆神经节 (英里), 以箭、海绵状神经 (CN) 为标志。<a href="/files/ftp_upload/56807/56807fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56807/56807fig6.jpg"> 图 6.海绵状神经的解剖.(A) 用微型剪刀切割海绵状神经上的筋膜。(B) 使用微钳将神经与底层组织分离。(C) 在神经下方放置9-0 结扎。主要骨盆神经节 (英里), 以箭、海绵状神经 (CN) 为标志。<a href="/files/ftp_upload/56807/56807fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/56807/56807fig7.jpg"> 图 7.挂钩的神经.(A) 通过轻轻拉缝线来提升神经。(B) 神经在电极的钩上休息。(C) 与生物相容的硅胶水隔离的神经和电极复合体。(D) 添加的附加胶水气泡完全隔离神经电极复合体。<a href="/files/ftp_upload/56807/56807fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/56807/56807fig8.jpg"> 图 8.插管的膜。23克针连接到 PE-50 油管插入到膜.用箭头标记的插入点。<a href="/files/ftp_upload/56807/56807fig8large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/56807/56807fig9.jpg"> 图 9.测试海绵行.用正确的行位置看到的响应。注意线的冲洗和对 cru 的攻丝响应。另外, 请注意快速压力降回基线。<a href="/files/ftp_upload/56807/56807fig9large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 10" class="xfigimg" src="/files/ftp_upload/56807/56807fig10.jpg"> 图 10.海绵状神经刺激的初步反应.减少了第一反应。首先刺激50毫米汞和一个起伏的高原. 二次和第三次刺激66毫米汞。以下测量记录在正常水平的73毫米汞.<a href="/files/ftp_upload/56807/56807fig10large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 11" class="xfigimg" src="/files/ftp_upload/56807/56807fig11.jpg"> 图 11.反复海绵神经刺激和海绵压力记录.使用此协议显示结果的稳定性。十的背部到背部刺激在75-78 毫米汞之间.<a href="/files/ftp_upload/56807/56807fig11large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 12" class="xfigimg" src="/files/ftp_upload/56807/56807fig12.jpg"> 图 12.海绵状神经刺激和压力记录.约30的背部和背部刺激与 < 6 毫米汞的变异性在压力。<a href="/files/ftp_upload/56807/56807fig12large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 13" class="xfigimg" src="/files/ftp_upload/56807/56807fig13.jpg"> 图 13.连续刺激持续20分钟最后增加的波动, 但随后的刺激, 在1分钟休息后, 产生了稳定的反应。<a href="/files/ftp_upload/56807/56807fig13large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 14" class="xfigimg" src="/files/ftp_upload/56807/56807fig14.jpg"> 图 14.泄漏膜.冲洗和攻丝后, 长时间响应回基线。刺激后反应减少。<a href="/files/ftp_upload/56807/56807fig14large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 15" class="xfigimg" src="/files/ftp_upload/56807/56807fig15.jpg"> 图 15.麻醉剂量对海绵压力的影响.相对于前两个2% 的稳定反应, 以及过去三的刺激反应, 对异氟醚增加的响应减少, 波动更大。上面的蓝色痕迹显示恒定平均动脉压。<a href="/files/ftp_upload/56807/56807fig15large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 16" class="xfigimg" src="/files/ftp_upload/56807/56807fig16.jpg"> 图 16.麻醉类型对海绵压力的影响.在异氟醚麻醉下进行的初始刺激显示80毫米汞的压力增加, 一旦芬太尼/咪达唑仑被管理, 对110毫米汞的反应增加.<a href="/files/ftp_upload/56807/56807fig16large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 17" class="xfigimg" src="/files/ftp_upload/56807/56807fig17.jpg"> 图 17.通过鼻锥的氧气管理的效果.通过鼻锥停止氧的管理, 导致海绵状神经刺激明显减少。未注意到平均动脉压的影响 (上面的蓝色痕迹)。<a href="/files/ftp_upload/56807/56807fig17large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 18" class="xfigimg" src="/files/ftp_upload/56807/56807fig18.jpg"> 图 18.电压对压力响应的影响.1.5-6 V 之间的电压产生了相同的压力响应。响应减少到 1.5 v 以下<a href="/files/ftp_upload/56807/56807fig18large.jpg" target="_blank">. 请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat at JoVE.com

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

本研究介绍了用硅胶胶和海绵压力测量分离神经电极复合物进行海绵状神经刺激的简化手术方法和技术。

大鼠海绵状神经刺激与海绵压力记录

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for ...

cavernous-nerve-stimulation-recording-intracavernous-pressure

Nanyang Technological University

Research

JoVE Journal

Medicine

12.4K 次观看.  University of Southern Denmark. 本研究介绍了用硅胶胶和海绵压力测量分离神经电极复合物进行海绵状神经刺激的简化手术方法和技术。

视频: Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

科研

医学

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

脑Microslices的兴奋性刺激作为<em&gt;在体外</em&gt;中风模型

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations over time in the blood-oxygenation level dependent signal. rs-fcMRI has been applied extensively to identify abnormalities in brain connectivity in different neurologic and psychiatric diseases. However, the relationship among rs-fcMRI connectivity abnormalities, brain electrophysiology and disease state is unknown, in part because the causal significance of alterations in functional connectivity in disease pathophysiology has not been established. Transcranial Magnetic Stimulation (TMS) is a technique that uses electromagnetic induction to noninvasively produce focal changes in cortical activity. When combined with electroencephalography (EEG), TMS can be used to assess the brain's response to external perturbations. Here we provide a protocol for combining rs-fcMRI, TMS and EEG to assess the physiologic significance of alterations in functional connectivity in patients with neuropsychiatric disease. We provide representative results from a previously published study in which rs-fcMRI was used to identify regions with abnormal connectivity in patients with epilepsy due to a malformation of cortical development, periventricular nodular heterotopia (PNH). Stimulation in patients with epilepsy resulted in abnormal TMS-evoked EEG activity relative to stimulation of the same sites in matched healthy control patients, with an abnormal increase in the late component of the TMS-evoked potential, consistent with cortical hyperexcitability. This abnormality was specific to regions with abnormal resting-state functional connectivity. Electrical source analysis in a subject with previously recorded seizures demonstrated that the origin of the abnormal TMS-evoked activity co-localized with the seizure-onset zone, suggesting the presence of an epileptogenic circuit. These results demonstrate how rs-fcMRI, TMS and EEG can be utilized together to identify and understand the physiological significance of abnormal brain connectivity in human diseases.

Resting-state functional-connectivity MRI has identified abnormalities in patients with a wide range of neuropsychiatric disorders, including epilepsy due to malformations of cortical development. Transcranial Magnetic Stimulation in combination with EEG can demonstrate that patients with epilepsy have cortical hyperexcitability in regions with abnormal connectivity.

评价中的连接有关的大脑兴奋患者癫痫多式联运Imaging-和激励为基础的方法

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations ...

Novel Approach for Simultaneous Recording of Renal Sympathetic Nerve Activity and Blood Pressure with Intravenous Infusion in Conscious, Unrestrained Mice.

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity (RSNA) is of great interest in many areas of research such as chronic kidney disease, hypertension, heart failure, diabetes and obesity. Unequivocal assessment of the role of the sympathetic nervous system is thus imperative for proper interpretation of experimental results and understanding of disease processes. RSNA has been traditionally measured in anesthetized rodents, including mice. However, mice usually exhibit very low systemic blood pressure and hemodynamic instability for several hours during anesthesia and surgery. Meaningful interpretation of RSNA is confounded by this non-physiological state, given the intimate relationship between sympathetic nervous tone and cardiovascular status. To address this limitation of traditional approaches, we developed a new method for measuring RSNA in conscious, freely-moving mice. Mice were chronically instrumented with radio-telemeters for continuous monitoring of blood pressure as well as a jugular venous infusion catheter and custom-designed bipolar electrode for direct recording of RSNA. Following a 48-72 hour recovery period, survival rate was 100% and all mice behaved normally. At this time-point, RSNA was successfully recorded in 80% of mice, with viable signals acquired up to 4 and 5 days post-surgery in 70% and 50% of mice, respectively. Physiological blood pressures were recorded in all mice (116&plusmn;2 mmHg; n=10). Recorded RSNA increased with eating and grooming, as well-established in the literature. Furthermore, RSNA was validated by ganglionic blockade and modulation of blood pressure with pharmacological agents. Herein, an effective and manageable method for clear recording of RSNA in conscious, freely-moving mice is described.

Anesthetized mice exhibit non-physiological systemic blood pressure, which precludes meaningful assessment of autonomic tone given the intimate relationship between blood pressure and the autonomic nervous system. Thus, a novel method to simultaneously record renal sympathetic nerve activity and blood pressure with intravenous infusion in conscious mice is outlined.

清醒、无拘无束的小鼠静脉输液同时记录肾交感神经活动和血压的新方法。

Renal sympathetic nerves contribute significantly to both physiological and pathophysiological phenomena. Evaluating renal sympathetic nerve activity ...

Subcutaneous Trigeminal Nerve Field Stimulation for Refractory Facial Pain

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. Neuromodulation techniques for chronic facial pain include deep brain stimulation and motor cortex stimulation, which are complex to perform. Subcutaneous nerve field stimulation is certified for chronic back pain and is the least invasive form of neuromodulation. We applied this technique to treat chronic and neuropathic trigeminal pain as an individual therapy concept. First, trial stimulation is performed. Subcutaneous leads are placed in the painful trigeminal dermatome under local anesthesia. The leads are connected to an external neurostimulator that applies constant stimulation. Patients undergo a 12 day outpatient trial to assess the effect of the stimulation. Electrodes are removed after the trial. If the patient reports pain reduction of at least 50% in intensity and/or attack frequency, a reduction in medication or increase in quality of life, permanent implantation is scheduled. New electrodes are implanted under general anesthesia and are subcutaneously tunneled to an infraclavicular internal pulse generator. Patients are able to turn stimulation on and off and to increase or decrease the stimulation amplitude as needed.
This technique represents a minimal invasive alternative to other more invasive means of neuromodulation for trigeminal pain such as motor cortex stimulation or deep brain stimulation.

Treating chronic or neuropathic facial pain can be challenging when medical or standard treatment fails. Subcutaneous nerve field stimulation is the least invasive form of neuromodulation and is used for chronic back pain. We applied this technology to treat chronic and neuropathic trigeminal facial pain.

皮下三叉神经场刺激难治性面部疼痛

Chronic or neuropathic trigeminal facial pain can be challenging to treat. Neurosurgical procedures should be applied when conservative treatment fails. ...

Induction and Micro-CT Imaging of Cerebral Cavernous Malformations in Mouse Model

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have been established by tamoxifen induced deletion of Ccm genes in postnatal animals. These mouse models provide invaluable tools to investigate molecular mechanism and therapeutic approaches for CCM disease. An accurate and quantitative method to assess lesion burden and progression is essential to harness the full value of these animal models. Here, we demonstrate the induction of CCM disease in a mouse model and the use of the contrast enhanced X-ray micro computed tomography (micro-CT) method to measure CCM lesion burden in mouse brains. At postnatal day 1 (P1), we used 4-hydroxytamoxifen (4HT) to activate Cre recombinase activity from the Cdh5-CreErt2 transgene to cleave the floxed allele of Ccm2. CCM lesions in mouse brains were analyzed at P8. For micro-CT, iodine based Lugol&#39;s solution was used to enhance contrast in brain tissue. We have optimized the scan parameters and utilized a voxel dimension of 9.5 &#181;m, which lead to a minimum feature size of approximately 25 &#181;m. This resolution is sufficient to measure CCM lesion volume and number globally and accurately, and provide high-quality 3-D mapping of CCM lesions in mouse brains. This method enhances the value of the established mouse models to study the molecular basis and potential therapies for CCM and other cerebrovascular diseases.

This protocol demonstrates the induction of cerebral cavernous malformation disease in a mouse model and uses contrast enhanced micro computed tomography to measure lesion burden. This method enhances the value of established mouse models to study the molecular basis and potential therapies for cerebral cavernous malformation and other cerebrovascular diseases.

小鼠脑海绵体畸形的诱导及显微 CT 成像

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have ...

Vagus Nerve Stimulation As an Adjunctive Neurostimulation Tool in Treatment-resistant Depression

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression (TRD). VNS exhibits antidepressive and stabilizing effects. This method is particularly useful as a long-term treatment, in which up to two-thirds of patients respond. The vagus nerve stimulator is positioned on the left vagus nerve during a surgical procedure and is activated telemetrically by a wand connected to a handheld computerized device. The treating physician can perform various adjustments of the vagus nerve stimulator during in-office visits (e.g., by modifying stimulation intensity or stimulation frequency) to achieve maximum therapeutic effects with low side effects. Set-up of the device usually takes several months. Typical side effects include wound infection, temporary salivation, coughing, paralysis of the vocal cords, bradycardia, or even asystole. The patient can stop the VNS by placing a magnet over the generator. The current protocol describes delivery of the specific stimulation tool and methods for adjusting the tuning parameters to achieve the best remission rates in patients with TRD.

Vagus nerve stimulation (VNS) has been shown to be effective as an adjunct treatment for treatment-resistant depression (TRD). VNS leads to antidepressive and antisuicidal effects and quality of life improvements. This protocol offers a step-by-step guide to managing and adjusting a vagus nerve stimulator for efficient treatment of TRD.

迷走神经刺激作为抗抑郁抑郁症的辅助神经刺激工具

Vagus nerve stimulation (VNS) is an approved neurostimulation therapy. The purpose of the method is to treat patients with therapy-resistant depression ...

Simultaneous Electrocardiography Recording and Invasive Blood Pressure Measurement in Rats

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. Research focusing on cardiovascular disease models, potential cardiovascular therapeutic targets or pharmaceutical agents requires assessment of systemic arterial pressure and heart rhythm changes. In situations where radio telemetry systems are not available or affordable, the technique of femoral artery cannulation is an alternative way to obtain intra-arterial pressure waveform recordings and systemic BP measurements. This technique is economical and can be performed with standard equipment in animal facilities. However, invasive arterial pressure recording requires cannulation of small arteries, which can be a challenging surgical skill. Here, we present step-by-step protocols for femoral artery cannulation procedures. Key procedures include the calibration of the data acquisition system, tissue dissection and femoral artery cannulation, and setup of the arterial cannulation system for pressure recording. Surface electrocardiography recording procedures are also included. We also present examples of BP recordings from normotensive and hypertensive rats. This protocol allows reliable direct recordings of systemic BP with simultaneous electrocardiography.

Here, we describe a setup for simultaneous recording of electrocardiography and intra-arterial blood pressure (BP) in experimental rats, which can be done with standard equipment in animal facilities and can be applied to physiological or pharmacological studies to investigate pathogenic or therapeutic mechanisms in cardiovascular medicine.

大鼠同时心电图记录及侵入性血压测量

For studies related to cardiovascular physiology or pathophysiology, blood pressure (BP) and electrocardiography are basic observational parameters. ...

Updated Technique for Reliable, Easy, and Tolerated Transcranial Electrical Stimulation Including Transcranial Direct Current Stimulation

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of brain stimulation presents several potential advantages compared to other techniques, as it is noninvasive, cost-effective, broadly deployable, and well-tolerated provided proper equipment and protocols are administered. Even though tDCS is apparently simple to perform, correct administration of the tDCS session, especially the electrode positioning and preparation, is vital for ensuring reproducibility and tolerability. The electrode positioning and preparation steps are traditionally also the most time consuming and error-prone. To address these challenges, modern tDCS techniques, using fixed-position headgear and pre-assembled sponge electrodes, reduce complexity and setup time while also ensuring that the electrodes are consistently placed as intended. These modern tDCS methods present advantages for research, clinic, and remote-supervised (at home) settings. This article provides a comprehensive step-by-step guide for administering a tDCS session using fixed-position headgear and pre-assembled sponge electrodes. This guide demonstrates tDCS using commonly applied montages intended for motor cortex and dorsolateral prefrontal cortex (DLPFC) stimulation. As described, selection of the head size and montage-specific headgear automates electrode positioning. Fully assembled pre-saturated snap-electrodes are simply affixed to the set position snap-connectors on the headgear. The modern tDCS method is shown to reduce setup time and reduce errors for both novice and expert operators. The methods outlined in this article can be adapted to different applications of tDCS as well as other forms of transcranial electrical stimulation (tES) such as transcranial alternating current stimulation (tACS) and transcranial random noise stimulation (tRNS). However, since tES is application specific, as appropriate, any methods recipe is customized to accommodate subject, indication, environment, and outcome specific features.

When administering transcranial direct current stimulation (tDCS), reproducible electrode preparation and placement are vital for a tolerated and effective session. The purpose of this article is to demonstrate updated modern setup procedures for the administration of tDCS and related transcranial electrical stimulation techniques, such as transcranial alternating current stimulation (tACS).

用于可靠、简单和耐受的颅内电刺激（包括颅内直流刺激）的更新技术

Transcranial direct current stimulation (tDCS) is a noninvasive method of neuromodulation using low-intensity direct electrical currents. This method of ...

大鼠海绵状神经刺激与海绵压力记录

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