Name: detecting establishment of shared blood supply in parabiotic mice by caudal vein glucose injection
Uploaded: 2020-02-06T12:30:00
Description: Watch this Scientific Journal Video about Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection at JoVE.com

这项工作得到了国家自然科学基金（81570399和81773735）、国家重点研究发展计划——中医药现代化研究项目（2017YFC1702003）和黑龙江的支持。杰出青年科学基金（JC2017020）。

所有涉及动物及其护理的程序均经哈尔滨医科大学动物护理与使用委员会批准。
注：该方法所需的工具和设备列在材料表中。
1. 材料和动物的制备
<ol><li>从标准实验室动物供应商处订购重量在 20 g-25 g 之间的 C57BL/6 雄性小鼠。</li>
	<li>将小鼠以12小时光的周期内房屋：在24~26°C下黑暗12小时，并可获取水和食物。</li>
</ol>2. 寄?...

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寄生虫学是指通过实验方法6、7、8连接两只活体动物建立共同血管系统的外科技术。该模型的优点是，单个个体产生的物质可以同时作用于两种动物。因此，寄生虫病模型可用于探索物质或因素在相关疾病中的作用，得出许多有意义的创新结论。鉴于其巨大的应用价值，该模型使人们对心血管系统疾病<sup clas...

寄生虫症是一种建模方法，其中两个活生物体通过手术连接，并发展成为一个与共享循环系统1的单一生理系统。由于单个个体产生的物质可以通过共享循环系统同时作用于两种动物，因此这些模型已广泛用于生理学研究。自19世纪中叶保罗·伯特2号开创的益生菌实验以来，构建生物模型的方法已经标准化。然而，一个简单方便的方法，以验证成功建立血性。据报道，交叉循环可以通过在其中一种副生物中注射0.5%埃文斯蓝色染料，然后用微孔板读取器3测量两种副生物血液中的Evans蓝色吸收率，从而成功地评估。另一种方法需要特定小鼠品种，其中包含 CD45.1+- 和 CD45.2=- 标记单核细胞在每个寄生虫中。然后，细胞细胞测定通过测量脾脏或血液4中单核细胞中的两个标记的频率来确定血液的血性。然而，这些方法往往对动物是致命的或繁琐的，并且非常需要一种安全而简单的方法来快速可靠地验证副生物模型。在这项研究中，我们建立了一种用于此目的的新方法，在寄生虫化小鼠模型中进行了验证。从尾静脉抽取的血液样本中的葡萄糖浓度使用胶质测定仪进行测量，供体小鼠和受体小鼠的葡萄糖水平变化模式被认为是循环性血球症的征兆。我们给这种方法命名为"葡萄糖波动法"。这种验证方法的应用并不仅限于小鼠，但可以扩展到不同的病理模型，但那些葡萄糖代谢严重调节不良的模型除外。该过程简单、省时且安全。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>curved forceps</td>
			<td>JZ surgical Instruments, China</td>
			<td>J31340</td>
			<td></td>
		</tr>
		<tr>
			<td>fine scissors</td>
			<td>JZ surgical Instruments, China</td>
			<td>WA1030</td>
			<td></td>
		</tr>
		<tr>
			<td>needle forcep</td>
			<td>JZ surgical Instruments, China</td>
			<td>60017961</td>
			<td></td>
		</tr>
		<tr>
			<td>2.5 mL syringes</td>
			<td>Agilent, USA</td>
			<td>5182-9642</td>
			<td></td>
		</tr>
		<tr>
			<td>Tribromoethano</td>
			<td>Sigma-Aldrich, USA</td>
			<td>T48402-5G</td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin</td>
			<td>Solarbio, China</td>
			<td>IP0150</td>
			<td></td>
		</tr>
		<tr>
			<td>tramadol</td>
			<td>Yijishiye, China</td>
			<td>YJT712520</td>
			<td></td>
		</tr>
		<tr>
			<td>glucometer</td>
			<td>Roche Diabetes Care, Indiana</td>
			<td>Accu-Chek Active test strips</td>
			<td></td>
		</tr>
		<tr>
			<td>Evan&#39;s blue Counterstain</td>
			<td>Solarbio, China</td>
			<td>G1810</td>
			<td></td>
		</tr>
		<tr>
			<td>depilatory cream</td>
			<td>Nair, USA</td>
			<td>LL9161</td>
			<td></td>
		</tr>
		<tr>
			<td>Warming Blanket (Heating pad)</td>
			<td>Kent Scientific Corp, USA</td>
			<td>TP-22G</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrical shaver</td>
			<td>Codos, China</td>
			<td>CP-5000</td>
			<td></td>
		</tr>
		<tr>
			<td>3-0,5-0 
			surgical suture</td>
			<td>Shanghai Medical Suture Needle Factory, China</td>
			<td>SYZ 3-0#, SYZ 5-0#</td>
			<td></td>
		</tr>
	</tbody>
</table>

detecting establishment of shared blood supply in parabiotic mice by caudal vein glucose injection

寄生虫症是一种实验方法，将两种平行动物沿着身体的纵轴进行手术结合。我们提出了一个方案，用于检测通过注射葡萄糖的胆热静脉在副生物中成功建立血性。建造了益生菌小鼠。通过尾静脉将葡萄糖注射到供体小鼠中，并在不同时间点使用血胶仪测量两只小鼠的血糖水平波动。结果表明，葡萄糖注射后，供体小鼠血糖水平在1分钟后急剧上升，此后缓慢下降。同时，受体小鼠的血糖水平在注射后15分钟达到峰值。与埃文斯蓝，用作葡萄糖的阳性对照获得类似的结果。血糖水平的同步波动表明两小鼠之间的血流成功。

在6只供体小鼠中，通过尾静脉注射100μL葡萄糖（1.2克/千克）后，血糖水平急剧上升至26.5μmol/L（173%增加），然后逐渐降至13.3μmol/L，60分钟。在受体小鼠中，注射后血糖缓慢升高，并在15分钟达到第一峰水平（增加47%，12.2μmol/L）。根据上述结果，循环血球化的标准设定如下：1）注射葡萄糖后1分钟内供体小鼠血糖水平急剧上升（至少增加100%或>20 μmol/L），2）注射后15分钟?...

Watch this Scientific Journal Video about Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection at JoVE.com

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

在这里，我们描述了一种新方法，通过注射葡萄糖，检测两种副生物的共享血液循环的成功建立，这种注射对副生子的损害最小，而且对寄生虫没有致命。

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 ...

本协议为循环奇体主义提供了一种改进的验证方法，也为生物医学研究成功建立寄生虫模型提供了一种方法。与现有的寄生虫病中血液志智检测方法相比，我们的协议既安全又简单，能够快速可靠地进行验证。虽然这种方法是特定在寄生虫病模型中检测血液千叶病的，但它可以应用于任何涉及寄生虫学程序的生物医学研究。最具挑战性的一步是葡萄糖注射。因此，在进行实验程序之前，请务必练习尾静脉注射，或使用静脉视觉小鼠尾部固定器。在确认对踏板反射缺乏反应后，将两只20至25克麻醉的C57黑色6雄性小鼠放在超膝位置，并用电剃须刀彻底剃光一只小鼠的左侧和第二只小鼠的右侧，从肘部上方约1厘米到膝盖以下一厘米。在将毛发膏去除任何剩余头发之前，先在每只动物的清除皮肤上切开纵向皮肤切口，从肘部上方 0.5 厘米到膝盖关节以下 0.5 厘米不等，然后轻轻地将皮肤从皮下筋膜上分离。用 3- 0 缝合连接副生物的奥勒克拉农和膝盖，用连续 5-0 缝合剃光的皮肤。然后皮下注射0.5毫升0.9%氯化钠以防止脱水，并在肌肉内每25克小鼠注射10毫克曲马多，以减轻疼痛，每天注射到每只小鼠三天。为了验证葡萄糖波动中副生物之间节流的成功构建，在副生物手术后的第10天，将供体小鼠固定在静脉视觉小鼠尾部固定器中，并使用70-75%乙醇浸泡的棉球摩擦一个副生物尾侧的果氏静脉。将一毫升注射器针头插入距离尾尖两到四厘米的胆碱静脉，并尽可能与静脉平行，在100微升盐水溶液中将每公斤1至2克葡萄糖注射到供体中，为期10秒。接下来用剪刀从供体和接受动物身上取下手趾，以便注射后在不同时间点收集一滴血。然后将每个血液样本的一滴添加到血糖仪测试条的中心，以检测葡萄糖水平。为了验证埃文斯蓝色染料吸收剂的循环成功构建，将200微升0.5%的埃文斯蓝色反面体注入供体小鼠，并在两小时后通过心脏穿刺从两个寄生虫中采集血液。通过离心分离血清，将血清转移到锥形管中，以0.9%氯化钠的1比50的比例稀释。然后在分光光度计上测量稀释血清样品在620纳米的吸光度。在这项对6只供体小鼠血糖水平的代表性分析中，在通过尾静脉注射100微升葡萄糖后，血糖急剧上升至每升26.5微摩尔。。血糖水平然后逐渐下降到13.3微升每升在60分钟。在接受小鼠中，注射后血糖缓慢升高，在15分钟内达到第一个峰值水平。Evans蓝色染料在准生物血清中的浓度也表明循环志结的成功构建，可以清楚地观察到副生物之间的皮下血管结。在寄生虫在寄生虫手术后15天没有建立血液反应的寄生虫中，接受小鼠在葡萄糖注射到供体动物后60分钟内没有表现出血糖水平升高。两个接受者中埃文斯蓝的血液浓度也未升高，表明葡萄糖波动法与埃文斯蓝法一样敏感。此外，由于葡萄糖水平迅速升高，在葡萄糖注射一小时后，血液胰岛素水平明显下降，并恢复到正常水平三小时，表明注射葡萄糖对胰岛素代谢的影响是可恢复的。小心确保针头直接插入尾静脉。在用这种方法确认准生物的血液囊反应后，可以研究活的准生物动物的生理病理变化。

Research

JoVE Journal

Biology

Drawing Blood from Rats through the Saphenous Vein and by Cardiac Puncture

从老鼠血液通过大隐静脉和心脏穿刺“

Drawing blood from rodents is necessary for a large number of both in vitro and in vivo studies. Sites of blood draws are numerous in rodents: ...

科研

生物学

Isolation and Culture of Pulmonary Endothelial Cells from Neonatal Mice

从新生小鼠的肺血管内皮细胞的分离和培养

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells ...

Detecting Somatic Genetic Alterations in Tumor Specimens by Exon Capture and Massively Parallel Sequencing

由外显子捕获和大规模平行测序检测体细胞遗传学改变在肿瘤标本

Efforts  to detect and investigate key oncogenic mutations have proven valuable to  facilitate the appropriate treatment for cancer patients. The ...

Identification of a Murine Erythroblast Subpopulation  Enriched in Enucleating Events by Multi-spectral Imaging Flow Cytometry

一个小鼠红细胞亚群富集去核活动由多光谱成像流式细胞仪鉴定

Erythropoiesis in mammals concludes with the dramatic process of enucleation that results in reticulocyte formation. The mechanism of enucleation has not ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Sampling Blood from the Lateral Tail Vein of the Rat

从大鼠的侧尾静脉血液取样

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, ...

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

糖皮质激素粪分析：在马属动物无创监测肾上腺

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

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 and Characterizing Protein Self-Assembly In Vivo by Flow Cytometry

通过流动细胞测定检测和描述 Vivo 中的蛋白质自组装

Protein self-assembly governs protein function and compartmentalizes cellular processes in space and time. Current methods to study it suffer from ...

Detecting Migration and Infiltration of Neutrophils in Mice

检测小鼠中性粒细胞的迁移和渗透

Neutrophils are a major member of the innate immune system and play pivotal roles in host defense against pathogens and pathologic inflammatory reactions. ...