我们要感谢 Kellie Breen Church 博士、Michael Kreisman 先生和 Jessica Jang 女士为收集代表性结果中显示的数据所做的贡献。这项工作得到了美国国立卫生研究院（NIH）R00 HD104994（R.B.M.）的支持。

所有程序均由科罗拉多州立大学 （#3960） 和加州大学圣地亚哥分校机构动物护理和使用委员会批准，并收集了代表性数据（S13235，PI Kellie Breen Church）。来自5只成年雌性和2只成年雄性C57 / BL6小鼠（9-16周龄）的数据在代表性数据部分中描述。如前所述，雌性小鼠在ICV注射和采血前3-4周进行卵巢切除术7。在实验之前，这些小鼠被饲养在12小时光照/ 12小时暗光循环中，并根据《实验动物护理和使用指南》自由获取饲料和水。
1. 进行开颅手术
注意：开颅手术可以在实际注射前一天或多天进行，这使得实验当天的注射过程更快。
<ol><li>如下所述准备开颅手术的材料。<ol><li>在工作台面上放置干净的台垫或悬垂材料。将异氟烷蒸发器的鼻锥粘在工作表面上（靠近实验者，但背对实验者;见 图1A）。</li>
		<li>将硅橡胶管放在无菌锋利的 18 G 针头的末端，使针尖突出 ~1 mm。将硅橡胶管放在无菌钝针的末端，使针尖突出~1毫米。 注意：每只小鼠应使用单独且无菌的预包装针头（锋利和钝）。</li>
		<li>收集电动剃须刀、碘磨砂膏、无菌纱布和酒精棉片，用于准备注射部位。</li>
	</ol></li>
	<li>如下所述，练习移动针头 2 毫米的横向和 1 毫米的尾部。<ol><li>用尺子和笔在一张纸上标记一个起点（这将是bregma），一个标记向左或向右2毫米，另一个标记在最后一个上方1毫米处（这将是注射部位;见 图1B）。</li>
		<li>练习将针头从起点移动到标记点 1 再到标记点 2（前囟到注射部位），直到确信该运动在没有导轨的情况下是可重复的。 注：此处描述的坐标在成年 C57/BL6 小鼠中有效，但其他品系或年龄组可能需要调整。</li>
	</ol></li>
	<li>如下所述准备小鼠进行开颅手术。<ol><li>将小鼠置于诱导室中，并在医用级氧气中用3%-4%异氟烷麻醉小鼠。通过练习和熟悉程序，将开颅手术的异氟烷暴露总持续时间减少到10分钟以内。 注意：异氟醚暴露对人体健康有害;在通风良好的地方使用经批准和检查的异氟烷汽化器，并清除废气。</li>
		<li>一旦脚趾反射消失，剃掉动物的头部。涂抹眼部润滑剂。</li>
		<li>将鼠标放在工作台面上，头部在鼻锥中以保持麻醉。</li>
		<li>按照供应商的指示施用镇痛剂，例如丁丙诺啡（0.6-0.8 mg/kg 小鼠体重，皮下注射）。</li>
		<li>用蘸有碘溶液的无菌纱布擦拭头部清洁注射部位（进行三次擦洗），然后用酒精擦洗垫擦拭（进行三次）。 注意：按照美国兽医协会的建议，去除毛发并清洁注射部位周围的皮肤，并使用无菌器械和针头，以防止感染。</li>
	</ol></li>
	<li>用非惯用手牢牢握住鼠标头部。将头部尽可能平放在工作台面上。</li>
	<li>找到注射部位。<ol><li>使用准备好的锋利的 18 G 针首先识别前盂;为此，将针沿着中线拖过头部皮肤，在喙尾平面上移动。想象一个等边三角形，其中两个顶点是眼睛，第三个顶点是囟的大致位置（见 图 1C）。</li>
		<li>从前乳开始，将针头向外侧移动2毫米，尾部向1毫米移动至注射部位。</li>
	</ol></li>
	<li>垂直握住针头的同时，用力将针头穿过皮肤和骨骼，直到管子与皮肤齐平。</li>
	<li>缩回针头，旋转针头，然后再次按压皮肤和骨骼。重复该过程，直到在骨头上形成一个小孔。</li>
	<li>使用钝针检查骨头上是否产生了足够的孔。目标是在骨头上开一个足够大的开口，以便钝针穿过。</li>
	<li>将鼠标从鼻锥中取出，用无菌纱布清除注射部位的任何血液，然后将鼠标放在笼子加热器上的笼子中，直到醒来。由于开颅手术的开口很小（18 G 针头），因此不需要用缝合线或伤口夹闭合该部位。 注意：这里产生的开口很小（18 G），因此许多机构认为这种手术是注射而不是手术。此外，虽然通过皮肤和骨骼向大脑开了一个口，但保持皮肤绷紧会导致孔不对齐，这可能有助于防止皮肤菌群或笼子垫料接触大脑。</li>
</ol>2. 进行注射
<ol><li>如下所述准备注射材料。<ol><li>在工作台面上放置干净的台垫或悬垂材料。将异氟烷蒸发器的鼻锥粘在工作表面上（靠近实验者，但背对实验者;见 图1A）。</li>
		<li>将一根 10 mm 长的 27 G 针头（斜角为 45°）连接到 5 μL 玻璃注射器上。将硅橡胶管放在针头上，使针尖突出 3.5 毫米;使用实验室胶带将管子固定在注射器主体上（见 图1D）。</li>
		<li>在试管中制备注射介质（药物、病毒或其他注射液）。对于本研究的代表性结果，注射无菌等渗盐水。 注意： 选择允许玻璃注射器和针头进入的试管，例如 2 mL 微量离心管。</li>
		<li>将 3 μL 注射介质吸入玻璃注射器中。首先，抽取超过所需体积的体积，然后弹出，直到注射器中残留 3 μL。</li>
		<li>收集碘磨砂膏、无菌纱布和酒精垫，用于准备注射部位。</li>
		<li>在计数模式下启动实验室计时器，并放置计时器，以便实验者在进样时可以看到它。</li>
		<li>练习像前一天一样移动针头 2 毫米的横向和 1 毫米的尾部。</li>
	</ol></li>
	<li>如下所述准备注射鼠标。<ol><li>将小鼠置于诱导室中，并用异氟醚麻醉小鼠。一旦脚趾反射消失，涂抹眼部润滑剂，然后将鼠标放在工作台面上，头部在鼻锥内。</li>
		<li>用碘和酒精各擦三片擦拭清洁注射部位。如步骤1.8所述，用18 G针（或钝针）识别注射部位。开颅手术期间形成的孔洞应该是可检测到的。 注意：如果没有提前进行开颅手术，或者如果无法检测到颅骨上的孔，则可以在此处进行开颅手术。</li>
	</ol></li>
	<li>如下所述用玻璃注射器进行注射。<ol><li>用非惯用手牢牢握住鼠标头部。将头部尽可能平放在工作台面上。</li>
		<li>将玻璃注射器的针头穿过头骨上的孔，直到放置在步骤2.1.2中制备的针头上的管子停留在小鼠的皮肤上。</li>
		<li>尽可能垂直握住注射器，注意冠状面和矢状面。在1分钟内缓慢注入培养基。</li>
		<li>注射完成后，将注射器和针头固定到位一分钟，以尽量减少回流。慢慢地将针头从鼠标头上缩回。</li>
		<li>将鼠标从鼻锥中取出，用无菌纱布（如果存在）清除注射部位的任何血液，然后将鼠标放在笼子加热器上的笼子中，直到醒来。</li>
	</ol></li>
</ol>3. 确认注射位置
<ol><li>如前所述，用盐水和4%多聚甲醛在磷酸盐缓冲盐水（PBS）中深度麻醉并灌注小鼠8。将大脑储存在4°C的PBS中的30%蔗糖中，直到大脑下沉（通常为2天）。</li>
	<li>如前所述，在低温恒温器上切割 20-50 μm 冠状切片9.切片时，观察组织块中的注射道。记录注射道是否与侧脑室明显相交。</li>
</ol>

使用徒手方法对小鼠进行脑室内注射

<ol>
	<li>Roseweir, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Discovery+of+potent+kisspeptin+antagonists+delineate+physiological+mechanisms+of+gonadotropin+regulation.">Discovery of potent kisspeptin antagonists delineate physiological mechanisms of gonadotropin regulation.</a> Journal of Neuroscience. 29 (12), 3920-3929 (2009).</li><li>Kim, J. Y., Grunke, S. D., Levites, Y., Golde, T. E., Jankowsky, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracerebroventricular+viral+injection+of+the+neonatal+mouse+brain+for+persistent+and+widespread+neuronal+transduction.">Intracerebroventricular viral injection of the neonatal mouse brain for persistent and widespread neuronal transduction.</a> Journal of Visualized Experiments. (91), e51863 (2014).</li><li>Taylor, Z. V., Khand, B., Porgador, A., Monsonego, A., Eremenko, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+optimized+intracerebroventricular+injection+of+CD4(+)+T+cells+into+mice.">An optimized intracerebroventricular injection of CD4(+) T cells into mice.</a> STAR Protocols. 2 (3), 100725 (2021).</li><li>Russo, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+control+of+the+female+reproductive+axis+through+gated+responsiveness+of+the+RFRP-3+system+to+VIP+signaling.">Circadian control of the female reproductive axis through gated responsiveness of the RFRP-3 system to VIP signaling.</a> Endocrinology. 156 (7), 2608-2618 (2015).</li><li>Laursen, S. E., Belknap, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracerebroventricular+injections+in+mice.+Some+methodological+refinements.">Intracerebroventricular injections in mice. Some methodological refinements.</a> Journal of Pharmacological Methods. 16 (4), 355-357 (1986).</li><li>Haley, T. J., McCormick, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+effects+produced+by+intracerebral+injection+of+drugs+in+the+conscious+mouse.">Pharmacological effects produced by intracerebral injection of drugs in the conscious mouse.</a> British Journal of Pharmacology and Chemotherapy. 12 (1), 12-15 (1957).</li><li>McCosh, R. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin-induced+hypoglycaemia+suppresses+pulsatile+luteinising+hormone+secretion+and+arcuate+Kiss1+cell+activation+in+female+mice.">Insulin-induced hypoglycaemia suppresses pulsatile luteinising hormone secretion and arcuate Kiss1 cell activation in female mice.</a> Journal of Neuroendocrinology. 31 (12), e12813 (2019).</li><li>Wu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transcardiac+perfusion+of+the+mouse+for+brain+tissue+dissection+and+fixation.">Transcardiac perfusion of the mouse for brain tissue dissection and fixation.</a> Bio-Protocol. 11 (5), e3988 (2021).</li><li>Comba, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+capture+microdissection+of+glioma+subregions+for+spatial+and+molecular+characterization+of+intratumoral+heterogeneity,+oncostreams,+and+invasion.">Laser capture microdissection of glioma subregions for spatial and molecular characterization of intratumoral heterogeneity, oncostreams, and invasion.</a> Journal of Visual Experiments. (158), e60939 (2020).</li><li>Porteous, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reformulation+of+PULSAR+for+analysis+of+pulsatile+LH+secretion+and+a+revised+model+of+estrogen-negative+feedback+in+mice.">Reformulation of PULSAR for analysis of pulsatile LH secretion and a revised model of estrogen-negative feedback in mice.</a> Endocrinology. 162 (11), (2021).</li><li>Hohmann, J. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+role+of+melanocortins+in+mediating+leptin's+central+effects+on+feeding+and+reproduction.">Differential role of melanocortins in mediating leptin's central effects on feeding and reproduction.</a> American Journal of Physiology: Regulatory, Integrative, and Comparative Physiology. 278 (1), R50-R59 (2000).</li><li>Gottsch, M. 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作者没有什么可透露的。

在这里，描述了一种在小鼠中进行ICV注射的简单有效的方法。由于该技术不需要立体定向框架，因此这种用于药物和实验剂集中递送的方法可供更多研究人员使用。此外，这种方法的通量相对较高，因为制备和注射程序可以快速执行。
由于此过程需要使用近似距离手动操作针头和玻璃注射器，因此建议新从业者在与活体动物一起工作之前花一些时间练习这些动作。此外，小?...

将实验药物（如药物1、病毒2 或细胞3）输送到大脑通常是神经内分泌研究所必需的。如果该药剂不容易穿过血脑屏障，或者实验目标是专门测试药剂的中枢作用，那么有一种可靠的方法将注射剂输送到大脑中是很重要的。此外，注射到脑室内 （ICV） 空间提供了将药物广泛分布在大脑中的机会，并提供了较大的靶区，从而增加了成功注射的可能性2. 
进行 ICV 注射的常用方法是放置永久性留置插管。在这种方法中，需要立体定位框架来定位市售或定制的套管，因为套管被粘合或胶合到位。通常，在康复后，通过套管给予超生理剂量的血管紧张素 II，如果立即观察到饮酒行为，则认为套管放置正确4.这种方法具有许多优点，包括能够进行长期输注和多次注射同一只动物的能力;此外，如果使用血管紧张素II，则可以在施用实验化合物之前确认正确的放置。然而，放置永久性插管存在一些局限性，包括对昂贵设备（立体定位框架）的要求、放置后插管损坏的可能性（例如，小鼠可以咀嚼笼子伴侣的插管）以及永久性插管周围感染的可能性。可以使用立体定位框架3 进行单次 ICV 注射，这虽然有效，但需要大量暴露于麻醉，因此可能会掩盖治疗的一些急性生理和行为影响。此外，将小鼠放置在立体定位框架中需要大量训练才能实现稳定放置并防止耳道破裂。
在这里，描述了一种在小鼠中进行徒手注射的既定方法。这种方法基于以前的报告5,6。这种技术的优点是简单、快速，不需要立体定位框架等专用设备。如下所述，该过程涉及操纵玻璃注射器相对于小鼠头部的标志进行注射，这可以快速完成，因此在实验当天只需要几分钟的气体麻醉。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>18-gauge blunt needles</td>
			<td>SAI Infusion</td>
			<td>B18-150</td>
			<td></td>
		</tr>
		<tr>
			<td>18-gauge needles</td>
			<td>BD Medical</td>
			<td>305195</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol pads</td>
			<td>Fisher Scientific</td>
			<td>22-363-750</td>
			<td></td>
		</tr>
		<tr>
			<td>Bench pad</td>
			<td>Fisher Scientific</td>
			<td>14-206-62AC22</td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine solution</td>
			<td>Fisher Scientific</td>
			<td>NC1696484</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Patterson Vet Supply</td>
			<td>07-892-5235</td>
			<td>Controlled substance</td>
		</tr>
		<tr>
			<td>Eyelube</td>
			<td>Fisher Scientific</td>
			<td>50-218-8442</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass syringe</td>
			<td>Hamilton</td>
			<td>7634-01</td>
			<td></td>
		</tr>
		<tr>
			<td>Injection needle</td>
			<td>Hamilton</td>
			<td>7803-01</td>
			<td>27 gauge, Small Hub RN needle, point style: 4, Needle length: 10cm, Angle: 45</td>
		</tr>
		<tr>
			<td>Isoflurane&#160;&#160;</td>
			<td>Patterson Vet Supply</td>
			<td>07-893-8441</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>Vet Equip</td>
			<td>V-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory Tape</td>
			<td>VWR</td>
			<td>89098-128</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical grade oxygen</td>
			<td>Airgas</td>
			<td>OX USPEA</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Millipore-Sigma</td>
			<td>8.18715.1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Fisher Scientific</td>
			<td>J67802.K2</td>
			<td></td>
		</tr>
		<tr>
			<td>PulsaR Software</td>
			<td>Open source, University of Otago</td>
			<td></td>
			<td>See ref 9</td>
		</tr>
		<tr>
			<td>Ruler</td>
			<td>Fisher Scientific</td>
			<td>12-00-152</td>
			<td></td>
		</tr>
		<tr>
			<td>Silastic tubing (0.040&#34; I.D.)</td>
			<td>DOW</td>
			<td>508-005</td>
			<td></td>
		</tr>
		<tr>
			<td>Silastic tubing (0.078&#34; I.D.)</td>
			<td>DOW</td>
			<td>508-009</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile saline</td>
			<td>VWR</td>
			<td>101320-574</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose&#160;</td>
			<td>Fisher Scientific</td>
			<td>S5-500</td>
			<td></td>
		</tr>
	</tbody>
</table>

free-hand intracerebroventricular injections in mice

神经内分泌系统的研究通常需要将药物、病毒或其他实验剂直接输送到小鼠的大脑中。脑室内 （ICV） 注射允许实验药物在整个大脑中广泛递送（特别是在心室附近的结构中）。在这里，描述了在成年小鼠中进行徒手ICV注射的方法。通过在小鼠头部使用视觉和触觉标志，可以快速可靠地向侧脑室注射。注射是用实验者手中的玻璃注射器进行的，并放置在与地标大致距离的地方。因此，该技术不需要立体定位框架。此外，该技术只需要短暂的异氟醚麻醉，这允许随后评估清醒、行为自由的小鼠的小鼠行为和/或生理学。徒手ICV注射是将实验剂有效输送到活体小鼠大脑中的强大工具，可以与其他技术相结合，如频繁的血液采样、神经回路操作或 体内 记录，以研究神经内分泌过程。

The delivery of experimental agents, such as drugs1, viruses2, or cells3, to the brain is often necessary for neuroendocrine research. If the agent does not readily cross the blood-brain barrier or the experimental objective is to specifically test the central effects of the agent, it is important to have a reliable method for delivering injections into the brain. Moreover, injection into the intracerebroventricular (ICV) space provides the opportunity to distribute the agent widely in the brain and provides a large target area, thus increasing the likelihood of successful injection2. 
A common method for making ICV injections involves the placement of a permanent indwelling cannula. In this approach, a stereotaxic frame is necessary to position the commercially available or custom-made cannula, as the cannula is glued or cemented in place. Often, upon recovery, a supraphysiological dose of angiotensin II is administered through the cannula, and if drinking behavior is immediately observed, then the cannula is considered correctly placed4. This approach has many advantages, including the ability to perform long-term infusion and the ability to inject the same animal multiple times; additionally, if angiotensin II is employed, correct placement can be confirmed prior to the administration of experimental compounds. However, there are some limitations to placing a permanent cannula, including the requirement for expensive equipment (stereotaxic frame), the possibility of damage to the cannula after placement (e.g., mice can chew on the cannula of a cage mate), and the possibility of infections around the permanent cannula. Single ICV injections can be made with the use of a stereotaxic frame3, which, although effective, requires substantial exposure to anesthesia and, thus, may obscure some acute physiological and behavioral effects of the treatment. Additionally, the placement of mice in a stereotaxic frame requires substantial training to achieve stable placement and prevent the rupturing of the ear canals.
Here, an established method for making free-hand injections in mice is described. This method is based on previous reports5,6. The advantages of this technique are that it is simple, rapid, and does not require specialized equipment such as a stereotaxic frame. As described below, this procedure involves manipulating a glass syringe relative to landmarks on the mouse head to make the injections, which can be done rapidly and, thus, requires only a few minutes of gas anesthesia on the experimental day.

作者聚焦：生殖神经控制的见解、创新和未来途径 

小鼠徒手脑室内注射

We are interested in the neural control of reproductive function, particularly how the brain regulates the release of hormones that control the gonads, both in healthy conditions and in response to stress. The main advantage of this technique is that it is relatively simple and rapid. Delivering drugs into the lateral ventricle is a great way to cast a wide net in testing a signaling pathway to support more detailed follow-up experiments.There's still much to be learned about the neuroendocrine control of reproduction, including the molecular mechanisms for luteinizing hormone pulse generation, mechanisms for modulating pulse frequency during stress, and generation of the preovulatory LH surge. In the future, we are particularly interested in integrating disparate findings into models.

成功执行后，该技术允许将实验剂快速输送到心室系统中。 图2A显示了接受ICV注射3μL无菌等渗盐水（许多药理学化合物的载体）的卵巢切除小鼠的促黄体激素（LH）脉冲曲线。该示例表明，短暂暴露于气体麻醉和单独向心室系统注射 3 μL 液体不会改变搏动性 LH 分泌。ICV注射后3 h，对动物实施安乐死，收集新鲜冷冻的神经组织，在低温恒温器上切割;注射道与侧脑室明显相交?...

Watch this Scientific Journal Video about Insights, Innovations, and Future Avenues in Reproductive Neurocontrol at JoVE.com

在这里，描述了一种使用徒手方法（即没有立体定位装置）在小鼠中进行脑室内注射的简单快速的方法。

The investigation of neuroendocrine systems often requires the delivery of drugs, viruses, or other experimental agents directly into the brains of mice. ...

我们对生殖功能的神经控制感兴趣，特别是大脑如何调节控制性腺的激素的释放，无论是在健康条件下还是在应对压力时。这种技术的主要优点是相对简单和快速。将药物输送到侧脑室是在测试信号通路以支持更详细的后续实验时广撒网的好方法。关于生殖的神经内分泌控制还有很多东西需要了解，包括促黄体生成素脉冲的分子机制、应激期间调节脉冲频率的机制以及排卵前 LH 激增的产生。未来，我们对将不同的发现整合到模型中特别感兴趣。

free-hand-intracerebroventricular-injections-in-mice

Research

JoVE Journal

Neuroscience

Free-Hand Intracerebroventricular Injections in Mice

6.5K 次观看.  Colorado State University. 我们对生殖功能的神经控制感兴趣，特别是大脑如何调节控制性腺的激素的释放，无论是在健康条件下还是在应对压力时。这种技术的主要优点是相对简单和快速。将药物输送到侧脑室是在测试信号通路以支持更详细的后续实验时广撒网的好方法。关于生殖的神经内分泌控制还有很多东西需要了解，包括促黄体生成素脉冲的分子机制、应激期间调节脉冲频率的机制以及?...

视频: 作者聚焦：生殖神经控制的见解、创新和未来途径 

The investigation of neuroendocrine systems often requires the delivery of drugs, viruses, or other experimental agents directly into the brains of mice. An intracerebroventricular (ICV) injection allows the widespread delivery of the experimental agent throughout the brain (particularly in the structures near the ventricles). Here, methods for making free-hand ICV injections in adult mice are described. By using visual and tactile landmarks on the heads of mice, injections into the lateral ventricles can be made rapidly and reliably. The injections are made with a glass syringe held in the experimenter&#39;s hand and placed at approximate distances from the landmarks. Thus, this technique does not require a stereotaxic frame. Furthermore, this technique requires only brief isoflurane anesthesia, which permits the subsequent assessment of mouse behavior and/or physiology in awake, freely behaving mice. Free-hand ICV injection is a powerful tool for the efficient delivery of experimental agents into the brains of living mice and can be combined with other techniques such as frequent blood sampling, neural circuit manipulation, or in vivo recording to investigate neuroendocrine processes.

Here, a simple and rapid approach for performing intracerebroventricular injections in mice using a free-hand approach (that is, without a stereotaxic device) is described.