Name: simple continuous glucose monitoring in freely moving mice
Uploaded: 2023-02-24T13:35:00
Description: Watch this Scientific Journal Video about Simple Continuous Glucose Monitoring in Freely Moving Mice at JoVE.com

我们感谢Dvir Mintz DVM和动物设施的兽医和畜牧工作人员以及我们小组的成员，进行了富有成效的讨论。这项研究得到了以色列科学基金会授予D.B.Z.的1541/21赠款的支持，D.B.Z.是Zuckerman STEM教师。

这些实验得到了希伯来大学机构动物护理和使用委员会（IACUC）的批准。
注意：所有工具都必须消毒，并且必须使用无菌技术处理套管。以下协议针对特定的 CGM 进行了微调。该协议可以适应其他CGM。
1.术前镇痛给药
<ol><li>皮下注射5%葡萄糖和0.45%生理盐水与美洛昔康，体重为5mg / kg。</li>
</ol>2.麻醉给药
<ol><li>...

<ol>
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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=Markers+of+glycemic+control+in+the+mouse:+Comparisons+of+6-h-and+overnight-fasted+blood+glucoses+to+Hb+A1c.">Markers of glycemic control in the mouse: Comparisons of 6-h-and overnight-fasted blood glucoses to Hb A1c.</a> American Journal of Physiology - Endocrinology and Metabolism. 295 (4), 981-986 (2008).</li><li>Xie, X., 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=Reduction+of+measurement+noise+in+a+continuous+glucose+monitor+by+coating+the+sensor+with+a+zwitterionic+polymer.">Reduction of measurement noise in a continuous glucose monitor by coating the sensor with a zwitterionic polymer.</a> Nature Biomedical Engineering. 2 (12), 894-906 (2018).</li><li>Van Der Meulen, T., 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=Urocortin3+mediates+somatostatin-dependent+negative+feedback+control+of+insulin+secretion.">Urocortin3 mediates somatostatin-dependent negative feedback control of insulin secretion.</a> Nature Medicine. 21 (7), 769-776 (2015).</li><li>Peterson, Q. P., 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=A+method+for+the+generation+of+human+stem+cell-derived+alpha+cells.">A method for the generation of human stem cell-derived alpha cells.</a> Nature Communications. 11, 2241 (2020).</li><li>Klueh, U., Liu, Z., Feldman, B., Kreutzer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interstitial+fluid+physiology+as+it+relates+to+glucose+monitoring+technologies:+Importance+of+Interleukin-1+and+Interleukin-1+receptor+antagonist+in+short-term+glucose+sensor+function+in+vivo.">Interstitial fluid physiology as it relates to glucose monitoring technologies: Importance of Interleukin-1 and Interleukin-1 receptor antagonist in short-term glucose sensor function in vivo.</a> Journal of Diabetes Science and Technology. 4 (5), 1073 (2010).</li><li>Klueh, U., Antar, O., Qiao, Y., Kreutzer, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+interleukin-1/interleukin-1+receptor+antagonist+family+of+cytokines+in+long-term+continuous+glucose+monitoring+in+vivo.">Role of interleukin-1/interleukin-1 receptor antagonist family of cytokines in long-term continuous glucose monitoring in vivo.</a> Journal of Diabetes Science and Technology. 7 (6), 1538 (2013).</li><li>Klueh, U., Kaur, M., Qiao, Y., Kreutzer, D. 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该协议提供了一种简单，廉价的方法来监测小鼠的葡萄糖水平，该方法不需要具有挑战性的显微手术，并且不涉及出血或处理小鼠。该方法易于在每个设施中实施，并且不会对小鼠造成死亡，疼痛或过度不适。协议中最关键的步骤是将葡萄糖传感器的套管插入小鼠的皮肤下。增加一些缝合线可以使套管在原位停留更长的时间。传感器很小，随着鼠标的移动，可能会被遮挡或放错位置。传感器必须?...

糖尿病（DM）是一种以高血糖水平为特征的毁灭性疾病。1型糖尿病可能是胰腺中产生胰岛素的β细胞的自身免疫攻击的结果。另一方面，2型DM和妊娠DM的特征在于β细胞无法分泌足够的胰岛素以响应葡萄糖水平的升高1。小鼠是用于研究DM的常见模式生物，因为它具有相似的生理机能，其正常葡萄糖水平接近人类。此外，由于关键信号通路的突变或暴露于特定饮食后，特定的小鼠品系可能会发展为DM，这使得疾病建模成为可能2，3，4。
通常使用为患者设计的血糖仪通过从小鼠尾巴尖端提取一小滴血液（1-2μL）来测量小鼠的血糖。这种方法会引起压力并需要处理小鼠，这会影响葡萄糖水平并禁止在自由行为的小鼠中或当研究人员不接近5时测量血糖水平。小鼠出血可能会对附近的小鼠造成压力，特别是对尚未测量血糖的同一笼子的小鼠，从而影响结果。小鼠的反应因处理者而异，测量葡萄糖的人可能会影响小鼠的葡萄糖水平。这些陷阱需要仔细的实验设计，并导致实验之间的一些不一致。
通过使用最先进的遥测技术将葡萄糖传感器植入小鼠的主动脉弓中，可以测量自由移动的小鼠的葡萄糖，而不会出血6。由此产生的测量结果非常好，可以持续很长时间，但是植入这些传感器具有挑战性，并且遥测系统价格昂贵，导致这种方法的适度采用，并且在非专业实验室中没有采用。近年来已经开发了针对小鼠尺寸及其生理机能量身定制的皮下或其他葡萄糖传感器，但这些再次需要高技能的专家，并且在某些情况下价格昂贵6，7，8，9，10。
最初开发用于监测DM患者血糖水平的商用连续血糖监测仪（CGM）提供了另一种在自由移动的小鼠中测量葡萄糖的选择，与植入探针相比，成本和技术专业知识要求更低。这些探针已被一些实验室5，11，12，13，14，15用于基础研究，包括使用该协议的同事16。这些设备通常包括传感器、安装设备、接收器和软件应用程序。传感器具有引导酶葡萄糖传感器的套管、胶带、能量源、短期记忆和存储数据并将其发送到接收器的无线通信模块。接收器可以显示当前的葡萄糖水平并将数据发送到服务器;这个接收器可以是手机。该软件应用程序为患者和医疗团队提供有关患者血糖的数据。在患者中，使用安装装置可以轻松连接传感器。通过将安装装置压在皮肤上来皮下插入套管，传感器在胶带的帮助下保持原位。
这是一个详细的协议，用于调整商业CGM设备以测量小鼠的葡萄糖水平。该协议描述了如何通过手术插入葡萄糖传感器并将其连接到鼠标。提供了用于基本数据分析和数据可视化的脚本。提供了潜在的陷阱、故障排除和标准结果的示例。以下协议特定于某个 CGM，但可以很容易地适应其他类型的商业 CGM。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2%&#160; Chlorhexidine Gluconate and 70%&#160; Isopropyl Alcohol</td>
			<td>3M</td>
			<td>ID&#160;7000136290</td>
			<td></td>
		</tr>
		<tr>
			<td>5% Dextrose and 0.45% Sodium Chloride Injection, USP</td>
			<td>Braun</td>
			<td>L6120</td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo needle holder</td>
			<td>FST</td>
			<td>12061-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Extra Fine Bonn scissors</td>
			<td>FST</td>
			<td>14084-08</td>
			<td></td>
		</tr>
		<tr>
			<td>FreeStyle Libre 1 reader</td>
			<td>Abbott</td>
			<td>ART27543&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>FreeStyle Libre sensor</td>
			<td>Abbott</td>
			<td>ART36687</td>
			<td></td>
		</tr>
		<tr>
			<td>FreeStyle Libre sensor applicator</td>
			<td>Abbott</td>
			<td>ART36787</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze pads</td>
			<td>Sion medical</td>
			<td>PC912017</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>FST</td>
			<td>11052-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair Removal Cream</td>
			<td>Veet</td>
			<td>3116523</td>
			<td></td>
		</tr>
		<tr>
			<td>High-fat high-sucrose diet</td>
			<td>Envigo Teklad diets</td>
			<td>TD.08811</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane, USP Terrell</td>
			<td>Piramal</td>
			<td>26675-46-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Meloxicam 5 mg/mL</td>
			<td>Chanelle Pharma</td>
			<td>08749/5024</td>
			<td></td>
		</tr>
		<tr>
			<td>MiniARCO Clipper kit</td>
			<td>Moser</td>
			<td>CL8787-KIT</td>
			<td></td>
		</tr>
		<tr>
			<td>PROLENE Polypropylene Suture 5-0</td>
			<td>Ethicon</td>
			<td>8725H</td>
			<td></td>
		</tr>
		<tr>
			<td>Puralube Opthalmic Ointment</td>
			<td>Perrigo</td>
			<td>574402511</td>
			<td></td>
		</tr>
		<tr>
			<td>Q-tips&#160;</td>
			<td>B.H.W</td>
			<td>271676</td>
			<td></td>
		</tr>
		<tr>
			<td>SomnoSuite Low-Flow Anesthesia System</td>
			<td>Kent Scientific</td>
			<td>SOMNO</td>
			<td></td>
		</tr>
	</tbody>
</table>

simple continuous glucose monitoring in freely moving mice

小鼠是用于研究糖尿病等代谢疾病的常见模式生物。葡萄糖水平通常通过尾出血来测量，这需要处理小鼠，引起压力，并且不能提供在黑暗周期中自由行为的小鼠的数据。在小鼠中进行最先进的连续葡萄糖测量需要将探针插入小鼠的主动脉弓以及专门的遥测系统。大多数实验室尚未采用这种具有挑战性且昂贵的方法。在这里，我们提出了一个简单的协议，涉及利用数百万患者使用的市售连续血糖监测仪来连续测量小鼠的葡萄糖，作为基础研究的一部分。葡萄糖感应探针通过皮肤的小切口插入小鼠背部的皮下空间，并使用几条缝合线紧紧固定到位。将设备缝合到鼠标皮肤上，以确保其保持在原位。该设备可以测量长达2周的葡萄糖水平，并将数据发送到附近的接收器，而无需处理小鼠。提供了用于记录的葡萄糖水平的基本数据分析的脚本。这种方法从手术到计算分析，具有成本效益，并且在代谢研究中可能非常有用。

手术结果 显示了八只HSD：ICR小鼠（8周龄）喂食高脂肪高蔗糖饮食（HFHS）18周和五只瘦HSD：ICR小鼠（12周龄）的结果。我们使用的设备存储数据长达 8 小时。进入当地动物设施的时间限制在07：00-19：00，因此禁止在小鼠活跃的下午晚些时候收集数据。因此，在手术前将小鼠放置在有反向照明的房间中7天，黑暗时间在8：30至20：30之间。并非所有设备或动物设施都需要这样做，我们建?...

Watch this Scientific Journal Video about Simple Continuous Glucose Monitoring in Freely Moving Mice at JoVE.com

Simple Continuous Glucose Monitoring in Freely Moving Mice

在这里，我们描述了一种简单的方法，将专为患者设计的商业连续血糖监测仪植入小鼠，并提供脚本来分析结果。

自由移动小鼠的简单连续血糖监测

Mice are a common model organism used to study metabolic diseases such as diabetes mellitus. Glucose levels are typically measured by tail-bleeding, which ...

我们的协议提供了一种简单廉价的方法来监测小鼠的葡萄糖水平。该技术的主要优点是它不需要具有挑战性的显微手术，并且不涉及出血或处理小鼠。该方法可用于鉴定肥胖和糖尿病小鼠模型中难以捉摸的葡萄糖管理表型。在尝试此方案时，最关键的步骤是将套管插入皮肤下，并收紧内部缝合线以保持套管就位。开始剃除麻醉小鼠背部的中线，并使用脱毛膏以确保完全脱毛。打开并加载传感器。然后将传感器安装到传感器安装设备上，露出传感器的胶带和套管侧。套管从针头上空无。将两个5-0锥点缝合到套管两侧的胶带上。对于背侧皮肤准备，使用锋利的剪刀在脊柱上方剃光区域的中心做一个两毫米的切口。在皮肤下短暂插入具有钝边的小镊子，形成一个小的皮下袋，以便套管可以很容易地插入口袋。将缝合线穿过切口两侧的皮肤。插入传感器时，请将传感器从传感器安装装置上完全取下，并用镊子握住传感器，以防止周围的胶带粘在自身上。小心地将套管插入皮下袋。拉动两侧的缝合线，然后拧紧并系好它们，将传感器牢固地固定到位。一旦胶带随着时间的推移松动，这将防止套管从皮下袋中滑出，切断缝合线的末端并丢弃它们。在传感器周围贴上胶带，并在传感器周围缝合八条不连续的缝合线，将传感器的边界贴在皮肤上。激活传感器。插入套管并成功激活 CGM 后，使用接收器收集数据。此处显示了使用提供的代码从单个鼠标获得的三天原始输出数据。轴参数和所有其他参数都可以在代码中调制。为了清楚起见，这里显示了三天的数据。制作一个皮下口袋并增加一些缝合线可以使传感器的套管在原位停留更长时间。使用该技术获得的连续数据使我们能够发现特定小鼠品系中难以捉摸的代谢表型。

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A Simple and Efficient Method for In Vivo Cardiac-specific Gene Manipulation by Intramyocardial Injection in Mice

A Simple and Efficient Method for In Vivo Cardiac-specific Gene Manipulation by Intramyocardial Injection in Mice

一种简便、有效的心肌注射小鼠心脏特异基因操作方法--体内

Gene manipulation specifically in the heart significantly potentiate the investigation of cardiac disease pathomechanisms and their therapeutic potential. ...

Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection

考达尔静脉葡萄糖注射剂在益生菌小鼠中检测共享血液供应的建立

Parabiosis is an experimental method for surgically combining two parallel animals along the longitudinal axis of the body. We present a protocol for ...

Simple Detection of Primary Cilia by Immunofluorescence

通过免疫荧光简单检测原发性西莉亚

Primary cilia are dynamically regulated during cell cycle progression, specifically during the G0/G1 phases of the cell cycle, being resorbed prior to ...