The authors would like to acknowledge the support of NIH/NIDDK K12 DK100014 (Lan Lu), the Case Comprehensive Cancer Center (NIH/NCI P30 CA43703), and the Clinical and Translation Science Collaborative of Cleveland (NIH/NCATS UL1 TR000439).

所有动物研究都是根据Case Western Reserve大学批准的机构动物护理和使用委员会（IACUC）方案进行的。下述外科手术的持续时间约为45-60分钟。每个时间点MRI程序本身约为15分钟。 麻醉<ol><li>在异氟烷麻醉室内麻醉大鼠，其设定为与氧气混合的2％异氟烷，以便在腹膜内施用可注射麻醉之前促进动物处理和约束。 </li><li>暴露于异氟烷3-5分钟后，检查动物是否被麻醉，并且对脚趾没有反应。 </li><li>进一步镇静大鼠腹膜内注射赛拉嗪和氯胺酮:75mg / kg氯胺酮/ 10mg / kg甲苯噻嗪。当进行腹膜内注射时，拉回针头以确保肠或其他重要器官的部分未被穿刺。 </li><li>一世在切口部位皮下注射2 mg / kg布比卡因以提供局部疼痛缓解。 </li></ol> 2.手术区域的准备<ol><li>在外科手术器械和用品用于手术之前灭菌并放置在外科手术垫上进行无菌处理。大多数仪器和耗材可以进行高压灭菌和重新使用。 </li><li>使用无菌手套进行所有外科手术。 </li><li>使用电动剃须刀将皮毛从动物的右侧剃掉。将动物从肋骨的底部剃除到后腿的顶部，为切口提供一个大的无毛的区域。 </li><li>将动物放在无菌手术垫上，以将消毒区域与周围区域隔离。 </li><li>用消毒剂如聚维酮碘或betadine擦洗皮肤。开始在手术中心的擦洗，并以圆形的方式移动到外面。用新的聚维酮碘或betadine擦拭至少重复三次。 </LI&gt; <li>用70％酒精拭子擦洗手术部位，直到皮肤清晰，因为如果吸收碘可能会有毒性。 </li></ol>手术程序<ol><li>在无菌条件下维持此程序。 </li><li>将麻醉动物置于左侧卧位的暖暖床上，右侧腹部朝上。 注意:必须注意将动物的核心体温保持在35-37°C以防止体温过低。这个升温床也应根据需要进行灭菌以维持无菌条件。 </li><li>感觉为肋骨笼，并使用无菌尺寸10手术刀刀片从肋骨底部开始，做一个小2-3厘米的右背腹膜后切口。 </li><li>沿切口两侧纵向放置无菌纱布。 </li><li>解剖皮下组织，脂肪和肌肉，以便可视化和进入腹腔。使用弯刀Mayo剪刀可以进一步穿透配给伤口并切割厚厚的组织。 </li><li>一旦肝脏清晰可见，便可使用钝镊子将肝脏向上收缩。 </li><li>另一方面使用另一对钝钳，暴露右肾，使其位于腹腔外。 </li><li>使用左手的手指和拇指将肾脏固定就位。用右手慢慢稳定地将0.1ml UTI89 大肠杆菌溶液（浓度在1×10 8 -1×10 9 ）从无菌注射器注入肾盂（其呈现为白色气泡） 15 。 注意:按照参考文献15所述准备细菌滴度。 </li><li>在针上放置一条可吸收的止血钳，以防止接种物流入腹膜。慢慢将针从肾盂拉出。 注意:注意不要刺穿或感染周围的组织导致非目标感染症状和/或并发症。 </li><li>使用较大的注射器，用生理盐水彻底冲洗肾脏，然后放回腹腔。 </li></ol>缝合线注意:将被埋在组织中的缝线应为4-0不可吸收的编织缝线。可吸收或单丝缝线可用于身体表面。 <ol><li>将缝合线均匀并尽可能靠近组织边缘，以防止血流阻塞;通常距离边缘不超过0.3厘米。 </li><li>抓住皮肤并使用一对细齿镊略微转动，并将针头保持器旋转到旋前的位置，以准备刺穿皮肤。 </li><li>通过放置手腕旋转针头并将其穿过皮肤，将针头穿过皮肤的全部厚度。 </li><li>对最接近执行过程的个人的皮肤边缘重复此过程。 </li><li>紧缩缝线足以抵抗组织边缘。任何更紧的会阻塞血液供应，伤口愈合缓慢，并可能导致开裂。 </li><li>使用方形结缝合缝合线，就好像它是一个简单的中断线迹，只不过只有短绞线被切割，留下约3-4毫米的尾巴。 </li><li>在第一针被束缚之后，准备放置距离第一针约3毫米的第二针，然后继续缝合线。 </li><li>一旦达到切口的末端，不要将最后一个针头完全拉过来。相反，使用在此处将针保持器保持的环作为短股线，以便将缝合线封闭件的远端扣紧。 </li><li>使用仪器领带，使用方形结缝合缝线。这导致3股从远端的完成的结粘连。 </li></ol> 5.动物恢复<ol><li>手术后腹膜内注射2.1 mg / kg育亨宾逆转麻醉。一个3-5小时后，预计手术完全恢复。 </li><li>手术后将动物放在加热垫（避免体温过低）和吸收性软床上。 </li><li>提供口服或肠胃外液体的补液，直到它能够恢复正常进食（24小时内）。手术后立即口服约0.6ml生理盐水溶液以限制脱水效果。 </li><li>按照IACUC批准的动物保健和使用协议中所述提供止痛。皮下注射5 mg / kg卡洛芬用于疼痛管理。 </li><li>定期监测切口是否肿胀，渗出，疼痛或开裂。 </li></ol> 6. 通过磁共振成像验证<ol><li>对高场小动物MRI扫描仪进行体内 MRI实验。 </li><li>用3％异氟烷在氧气中诱导麻醉，并将动物与右肾在等中心放置在MRI扫描仪中吃射频线圈。在整个成像过程中通过鼻锥连续地给动物1-2％的异氟烷​​麻醉。 注意:对于图1所示的体内 MRI图像，使用大鼠体积线圈（内径= 72mm）。 </li><li>使用动物监测和控制系统来维持每只动物的呼吸速率（40-60次呼吸/分钟）和核心体温（35±1℃）。 </li><li>使用多层，多回波自旋回波MRI采集获得感染和对照肾脏的高分辨率，轴向T2加权图像。典型的MRI采集参数是重复时间= 5000msec，回波时间= 40msec，切片厚度= 2.0mm，平面内空间分辨率=200μm，3个信号平均值，并且采集时间为8分钟。 </li></ol>

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The authors have no other disclosures.

啮齿动物（即小鼠和大鼠）升高的急性肾盂肾炎可以通过经尿道导尿术产生。 16，17，18经尿道这种感染的方法是有利的，因为它是非侵入性的和模拟上行感染的人类病理生理学。 17，18，19，20然而，这种方法也可以遭受不?...

啮齿动物模型已被用于研究许多人类疾病表现，包括肾盂肾炎和尿路感染（UTI）。 UTI是一个全球性的健康问题，可以影响所有年龄段的儿童，男性和女性。 1，2，3尿路感染的初始表现形式包括膀胱炎，如果感染沿输尿管上升，肾脏感染（肾盂肾炎）可以遵循。与此同时，全球糖尿病患病率接近四亿人。 4,5重要的是，尿路感染发病率可能高达谁是病人肥胖或有2型糖尿病，导致复发性尿路感染感染（入替），败血症，从肾盂肾炎肾纤维化和膀胱功能障碍的风险增加4倍以上升高。 6，7，8鼠类模型在研究UTI方面很重要，因为目前的抗生素疗法仅在UTI患者的一个子集中产生持续的预防性应答。为了改善临床UTI护理，关键步骤是了解rUTI及其从急性感染到炎症到纤维化的病理生理过程的机制以及2型糖尿病的影响。 改进动物模型的目标是开发可以更准确地评估疾病进展和治疗干预措施的技术。已经采用几种不同的方法在大鼠和/或小鼠中诱导肾盂肾炎，以研究肾损伤的病理生理学，抗生素治疗的效果以及UTI的自然过程的其他方面。建立逆行性UTI的常见方法是经尿道导尿。 10，11，12，13 &lt;/ sup&gt;该方法通过尿道将细菌引入麻醉动物的膀胱。虽然这种技术密切模拟人类肾盂肾炎，但由于多种因素，肾盂肾炎感染的实际发生率和大小可能是非常可变的，包括在接种期间或之后紧随其后的输尿管反流自发或排尿不足。 11其结果是，在诱导上行肾盂肾炎的感染可以限制这种模型来研究肾脏感染以及治疗策略的效用的实验变化。 该报告描述了一种手术肾盂肾炎大鼠模型，其中将大肠杆菌直接注射入大鼠肾脏。尽管这种大鼠模型是侵入性的，但是可以有效地控制输送到肾脏的大肠杆菌的数量，从而能够强壮的肾脏感染和炎症。 14在这个过程中，我们还描述如何通过体内磁共振成像（MRI）纵向监测这些诱导的肾脏感染。

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			<td height="21" style="height:21px;">Isoflurane</td>
			<td>Henry Schein Animal Health</td>
			<td>050033</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazine</td>
			<td>Henry Schein Animal Health</td>
			<td>33197</td>
			<td>Inject IP</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ketamine</td>
			<td>Patterson Vetrinary&#160;</td>
			<td>07-881-9413</td>
			<td>Inject IP</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Yohimbine (Atipamezole)</td>
			<td style="width:195px;">Patterson Vetrinary&#160;</td>
			<td>07-867-7097</td>
			<td>Inject IP after surgery</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bupivacaine (Marcaine)</td>
			<td style="width:195px;">Patterson Vetrinary&#160;</td>
			<td>07-890-4584</td>
			<td>Inject SQ at site of incision&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Carprofen (Rimadyl)</td>
			<td style="width:195px;">Patterson Vetrinary&#160;</td>
			<td>07-844-7425</td>
			<td>Should be kept at 4 &#7506;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">4-0 Chromic Gut Suture</td>
			<td style="width:195px;">Ethicon Inc.</td>
			<td>U203H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4-0 Braided Vicryl Suture</td>
			<td>Ethicon Inc.</td>
			<td>J304H</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1mL SubQ Syringe</td>
			<td>BD Biosciences</td>
			<td>309597</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">E. coli&#160; UTI89 or CFT073</td>
			<td style="width:195px;">ATCC</td>
			<td style="width:168px;">700928</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Surgicel Absorbable Hemostat</td>
			<td style="width:195px;">Ethicon Inc.</td>
			<td>ETH1951CS&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biospec 9.4T MRI&#160;</td>
			<td style="width:195px;">Bruker&#160;</td>
			<td style="width:168px;">94/20 USR</td>
			<td></td>
		</tr>
	</tbody>
</table>

intrarenal injection of escherichia coli in a rat model of pyelonephritis

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal inflammation and fibrosis ultimately resulting in declining kidney function. Before improved clinical management and prevention of pyelonephritis can be instituted, a reliable animal model must be established in order to study the mechanisms of progression, recurrence, and therapeutic efficacy. The transurethral infection model closely mimics human pyelonephritis but exhibits considerable variation due to its reliance on urethral reflux to transport the bacteria to the kidney. Herein, a detailed surgical protocol for performing bacterial injections into the rat renal pelvis is provided and confirmed by non-invasive Magnetic Resonance Imaging (MRI). Using this protocol, animals receive direct exposure to a desired concentration of E. coli bacteria and can fully recover from the surgical procedure with adequate post-operative care. This facilitates subsequent longitudinal MRI assessments of the experimental animal models for comparison with saline (sham) controls. Using this direct delivery approach, the severity of infection is controllable and applicable for mechanistic studies of progression as well as development of novel treatment strategies.

医学成像技术提供了非侵入性评估UTI和治疗功效的机会。因此，MRI用于验证注射1-2×10 7 UTI89 大肠杆菌后的急性感染诱导，并可视化手术前后肾脏的变化。 图1a-b显示肾脏感染逐渐增加的区域（黄色箭头）。在感染后第1天和第4天为每只动物获得的MRI图像有助于表征急性肾盂肾炎感染的生长。 <img alt="?...

Watch this Scientific Journal Video about Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis at JoVE.com

Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

This manuscript describes a rat surgical model of pyelonephritis using direct intra-renal infection by Escherichia coli into the renal pelvis. The experimental procedure can be utilized to study the pathogenesis of pyelonephritis as well as the associated inflammation and fibrosis.

肾内注射<em&gt;大肠杆菌</em&gt;在肾性肾炎大鼠模型中

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal ...

intrarenal-injection-escherichia-coli-rat-model

Research

JoVE Journal

Medicine

Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

7.9K 次观看.  Case Western Reserve University. This manuscript describes a rat surgical model of pyelonephritis using direct intra-renal infection by Escherichia coli into the renal pelvis. The experimental procedure can be utilized to study the pathogenesis of pyelonephritis as well as the associated inflammation and fibrosis.

视频: Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo. With the advent of genetic modifications to the whole organism or the myocardium and an array of biological and/or synthetic materials, there is great potential for any combination of these to assuage the extent of acute ischemic injury and impede the onset of heart failure pursuant to myocardial remodeling. 
Here we present the methods and materials used to reliably perform this microsurgery and the modifications involved for temporary (with reperfusion) or permanent coronary artery occlusion studies as well as intramyocardial injections. The effects on the heart that can be seen during the procedure and at the termination of the experiment in addition to histological evaluation will verify efficacy. 
Briefly, surgical preparation involves anesthetizing the mice, removing the fur on the chest, and then disinfecting the surgical area. Intratracheal intubation is achieved by transesophageal illumination using a fiber optic light. The tubing is then connected to a ventilator. An incision made on the chest exposes the pectoral muscles which will be cut to view the ribs. For ischemia/reperfusion studies, a 1 cm piece of PE tubing placed over the heart is used to tie the ligature to so that occlusion/reperfusion can be customized. For intramyocardial injections, a Hamilton syringe with sterile 30gauge beveled needle is used. When the myocardial manipulations are complete, the rib cage, the pectoral muscles, and the skin are closed sequentially. Line block analgesia is effected by 0.25% marcaine in sterile saline which is applied to muscle layer prior to closure of the skin. The mice are given a subcutaneous injection of saline and placed in a warming chamber until they are sternally recumbent. They are then returned to the vivarium and housed under standard conditions until the time of tissue collection. At the time of sacrifice, the mice are anesthetized, the heart is arrested in diastole with KCl or BDM, rinsed with saline, and immersed in fixative. Subsequently, routine procedures for processing, embedding, sectioning, and histological staining are performed. 
Nonsurgical intubation of a mouse and the microsurgical manipulations described make this a technically challenging model to learn and achieve reproducibility. These procedures, combined with the difficulty in performing consistent manipulations of the ligature for timed occlusion(s) and reperfusion or intramyocardial injections, can also affect the survival rate so optimization and consistency are critical.

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

结扎冠状动脉内注射在小鼠模型梗死

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

科研

医学

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

利用磁共振成像，容积电脑断层扫描及超音波的裸鼠乳腺癌骨转移大鼠模型血管生成的多模态成像

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Rat Model of Blood-brain Barrier Disruption to Allow Targeted Neurovascular Therapeutics

Endothelial cells with tight junctions along with the basement membrane and astrocyte end feet surround cerebral blood vessels to form the blood-brain barrier1. The barrier selectively excludes molecules from crossing between the blood and the brain based upon their size and charge. This function can impede the delivery of therapeutics for neurological disorders. A number of chemotherapeutic drugs, for example, will not effectively cross the blood-brain barrier to reach tumor cells2. Thus, improving the delivery of drugs across the blood-brain barrier is an area of interest.
The most prevalent methods for enhancing the delivery of drugs to the brain are direct cerebral infusion and blood-brain barrier disruption3. Direct intracerebral infusion guarantees that therapies reach the brain; however, this method has a limited ability to disperse the drug4. Blood-brain barrier disruption (BBBD) allows drugs to flow directly from the circulatory system into the brain and thus more effectively reach dispersed tumor cells. Three methods of barrier disruption include osmotic barrier disruption, pharmacological barrier disruption, and focused ultrasound with microbubbles. Osmotic disruption, pioneered by Neuwelt, uses a hypertonic solution of 25% mannitol that dehydrates the cells of the blood-brain barrier causing them to shrink and disrupt their tight junctions. Barrier disruption can also be accomplished pharmacologically with vasoactive compounds such as histamine5 and bradykinin6. This method, however, is selective primarily for the brain-tumor barrier7. Additionally, RMP-7, an analog of the peptide bradykinin, was found to be inferior when compared head-to-head with osmotic BBBD with 25% mannitol8. Another method, focused ultrasound (FUS) in conjunction with microbubble ultrasound contrast agents, has also been shown to reversibly open the blood-brain barrier9. In comparison to FUS, though, 25% mannitol has a longer history of safety in human patients that makes it a proven tool for translational research10-12.
In order to accomplish BBBD, mannitol must be delivered at a high rate directly into the brain's arterial circulation. In humans, an endovascular catheter is guided to the brain where rapid, direct flow can be accomplished. This protocol models human BBBD as closely as possible. Following a cut-down to the bifurcation of the common carotid artery, a catheter is inserted retrograde into the ECA and used to deliver mannitol directly into the internal carotid artery (ICA) circulation. Propofol and N2O anesthesia are used for their ability to maximize the effectiveness of barrier disruption13. If executed properly, this procedure has the ability to safely, effectively, and reversibly open the blood-brain barrier and improve the delivery of drugs that do not ordinarily reach the brain 8,13,14.

Blood-brain barrier disruption aids the delivery of certain drugs to the brain. Mannitol delivered intra-arterially shrinks cells surrounding blood vessels in order to physically disrupt the barrier.

允许有针对性的大鼠模型血 - 脑屏障破坏神经血管治疗

Endothelial cells with tight junctions along with the basement membrane and astrocyte end feet surround cerebral blood vessels to form the blood-brain ...

A Model of Chronic Nutrient Infusion in the Rat

Chronic exposure to excessive levels of nutrients is postulated to affect the function of several organs and tissues and to contribute to the development of the many complications associated with obesity and the metabolic syndrome, including type 2 diabetes. To study the mechanisms by which excessive levels of glucose and fatty acids affect the pancreatic beta-cell and the secretion of insulin, we have established a chronic nutrient infusion model in the rat. The procedure consists of catheterizing the right jugular vein and left carotid artery under general anesthesia; allowing a 7-day recuperation period; connecting the catheters to the pumps using a swivel and counterweight system that enables the animal to move freely in the cage; and infusing glucose and/or Intralipid (a soybean oil emulsion which generates a mixture of approximately 80% unsaturated/20% saturated fatty acids when infused with heparin) for 72 hr. This model offers several advantages, including the possibility to finely modulate the target levels of circulating glucose and fatty acids; the option to co-infuse pharmacological compounds; and the relatively short time frame as opposed to dietary models. It can be used to examine the mechanisms of nutrient-induced dysfunction in a variety of organs and to test the effectiveness of drugs in this context.

A protocol for chronic infusions of glucose and Intralipid in rats is described. This model can be used to study the impact of nutrient excess on organ function and physiological parameters.

在大鼠模型慢性营养输液

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

Development of an Alpha-synuclein Based Rat Model for Parkinson's Disease via Stereotactic Injection of a Recombinant Adeno-associated Viral Vector

In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on animal models. The identification of PD-associated genes has led to the development of genetic PD models. Most transgenic &#945;-SYN mouse models develop gradual &#945;-SYN pathology but fail to display clear dopaminergic cell loss and dopamine-dependent behavioral deficits. This hurdle was overcome by direct targeting of the substantia nigra with viral vectors overexpressing PD-associated genes. Local gene delivery using viral vectors provides an attractive way to express transgenes in the central nervous system. Specific brain regions can be targeted (e.g. the substantia nigra), expression can be induced in the adult setting and high expression levels can be achieved. Further, different vector systems based on various viruses can be used. The protocol outlines all crucial steps to perform a viral vector injection in the substantia nigra of the rat to develop a viral vector-based alpha-synuclein animal model for Parkinson&#39;s disease.

This manuscript describes how viral vector-mediated local gene delivery provides an attractive way to express transgenes in the central nervous system. The protocol outlines all crucial steps to perform a viral vector injection in the substantia nigra of the rat to develop a viral vector-based animal model for Parkinson's disease.

通过重组腺相关病毒载体注射立体的α-突触核蛋白基于大鼠模型为帕金森氏病的发展

In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Acupuncture in a Rat Model of Asthma

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes methods for the fabrication of the high platform, establishes a rat model of asthma and measures changes in respiratory function using a noninvasive and real-time whole-body plethysmography (WBP) system.

哮喘鼠模型中的针灸

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

Retinal Pathophysiological Evaluation in a Rat Model

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal detachment, breakdown of the blood-retinal barrier (BRB), and retinal angiogenesis. An in vivo rat model is a valuable experimental tool to examine the changes in the structure and function of the retina. We propose three different experimental techniques in the rat model to identify morphological changes of retinal cells, retinal vasculature, and compromised BRB. Retinal histology is used to study the morphology of various retinal cells. Also, quantitative measurement is performed by retinal cell count and thickness measurement of different retinal layers. A BRB breakdown assay is used to determine the leakage of extraocular proteins from the plasma to vitreous tissue due to the breakdown of BRB. Fluorescence angiography is used to study angiogenesis and leakage of blood vessels by visualizing retinal vasculature using FITC-dextran dye.

Diabetic retinopathy is one of the leading causes of blindness. Histology, blood-retinal barrier breakdown assay, and fluorescence angiography are valuable techniques to understand the pathophysiology of the retina, which could further enhance the efficient drug screening against diabetic retinopathy.

大鼠模型中的视网膜病理生理学评估

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal ...

Development of a Uterosacral Ligament Suspension Rat Model

Pelvic organ prolapse (POP) is a common pelvic floor disorder (PFD) with the potential to significantly impact a woman's quality of life. Approximately 10%-20% of women undergo pelvic floor repair surgery to treat prolapse in the United States. PFD cases result in an overall $26.3 billion annual cost in the United States alone. This multifactorial condition has a negative impact on the quality of life and yet the treatment options have only dwindled in the recent past. One common surgical option is uterosacral ligament suspension (USLS), which is typically performed by affixing the vaginal vault to the uterosacral ligament in the pelvis. This repair has a lower incidence of complications compared to those with mesh augmentation, but is notable for a relatively high failure rate of up to 40%. Considering the lack of standard animal models to study pelvic floor dysfunction, there is an urgent clinical need for innovation in this field with a focus on developing cost-effective and accessible animal models. In this manuscript, we describe a rat model of USLS involving a complete hysterectomy followed by fixation of the remaining vaginal vault to the uterosacral ligament. The goal of this model is to mimic the procedure performed on women to be able to use the model to then investigate reparative strategies that improve the mechanical integrity of the ligament attachment. Importantly, we also describe the development of an in situ tensile testing procedure to characterize interface integrity at chosen time points following surgical intervention. Overall, this model will be a useful tool for future studies that investigate treatment options for POP repair via USLS.

Pelvic organ prolapse affects millions of women worldwide and yet some common surgical interventions have failure rates as high as 40%. The lack of standard animal models to investigate this condition impedes progress. We propose the following protocol as a model for uterosacral ligament suspension and in vivo tensile testing.

子宫骶韧带悬浮大鼠模型的开发

Pelvic organ prolapse (POP) is a common pelvic floor disorder (PFD) with the potential to significantly impact a woman's quality of life. Approximately ...

A Model of Long-Term Ventricular Fibrillation in Isolated Rat Hearts

Ventricular fibrillation (VF) is a fatal arrhythmia with a high incidence in cardiac patients, but VF arrest under perfusion is a neglected method of intraoperative arrest in the field of cardiac surgery. With recent advances in cardiac surgery, the demand for prolonged VF studies under perfusion has increased. However, the field lacks simple, reliable, and reproducible animal models of chronic ventricular fibrillation. This protocol induces long-term VF through alternating current (AC) electrical stimulation of the epicardium. Different conditions were used to induce VF, including continuous stimulation with a low or high voltage to induce long-term VF and stimulation for 5 min with a low or high voltage to induce spontaneous long-term VF. The success rates of the different conditions, as well as the rates of myocardial injury and recovery of cardiac function, were compared. The results showed that continuous low-voltage stimulation induced long-term VF and that 5 min of low-voltage stimulation induced spontaneous long-term VF with mild myocardial injury and a high rate of recovery of cardiac function. However, the low-voltage, continuously stimulated long-term VF model had a higher success rate. High-voltage stimulation provided a higher rate of VF induction but showed a low defibrillation success rate, poor recovery of cardiac function, and severe myocardial injury. On the basis of these results, continuous low-voltage epicardial AC stimulation is recommended for its high success rate, stability, reliability, reproducibility, low impact on cardiac function, and mild myocardial injury.

This protocol presents a model of long-term ventricular fibrillation in rat hearts induced by continuous stimulation with low-voltage alternating current. This model has a high success rate, is stable, reliable, and reproducible, has a low impact on cardiac function, and causes only mild myocardial injury.

孤立大鼠心脏长期心室颤动模型

Ventricular fibrillation (VF) is a fatal arrhythmia with a high incidence in cardiac patients, but VF arrest under perfusion is a neglected method of ...

肾内注射

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