Name: protocol for long duration whole body hyperthermia in mice
Uploaded: 2012-08-25T12:00:00.000+00:00
Duration: 7 min 56 s
Description: Watch this Scientific Journal Video about Protocol for Long Duration Whole Body Hyperthermia in Mice at JoVE.com

1。动物伦理在执行长持续时间热疗对小鼠确保机构动物管理和使用委员会已批准的程序。本文讨论的所有程序，并显示在影随行的国家免疫学研究所实验动物伦理委员会的批准。 2。了解热疗全身热疗（WBH）被传递放置在预热的网箱孵化器被放置在保持在39.5℃下的动物在标准化的协议，一个温度传感器，在小鼠体内的外科手术植入，并在不同的实验条件下监测其体温。据观察，在热环境下的动物时，它们的温度逐渐上升至37°C到42°C，持续时间为100分钟（ 图1）。钍Ë产生的"秒杀"的温度是耐人寻味的，因为身体内部的温度射门甚至高于外部环境温度。 当身体温度超过40℃，动物将显示如润湿口鼻部周围的应力和脱水的迹象。增加身体的温度超过40°C，会导致润湿性和不可逆的压力，甚至死亡的动物，由于热应力。因此，重要的是要防止在热疗过程中的温度峰值。 这种高温尖峰现象关联与物理亢进的动物的在热疗曝光期间。相关温度峰值由其他组中观察到过，并与的动物，在被引入到环境中具有更高的温度下的10的物理多动症。 在我们的协议中，小鼠被固定在热疗曝光。如如图2所示，固定化的动物的防止温度峰值。 我们用8-12周龄BALB / cJ小鼠，证明此协议。该协议并不限于一个特定的类型/应变的小鼠。 3。实验准备 3.1孵化器该实验可在一个正常的CO 2培养箱中进行，因为它们具有良好的温度控制。断开CO 2气体的供给和除去水的CO 2培养箱中保持托盘。在某些型号的孵化器中，它是可以关掉的CO 2控制器。安排货架上的孵化器，一个老鼠笼可以方便放在里面的。孵化器的温度设定在39.5℃。 3.2网箱通常，热疗进行实验，在一个时间上的4只小鼠。两笼;e被置于孵化器内的，另一种是在室温下放在外面。床上用品的笼子是可以避免的，因为随着时间的推移，这将导致非均匀温度在笼子里面的床上用品变湿的补丁。外面的笼子应该有食物颗粒和一瓶水。 3.3小鼠固定管在热疗小鼠的第4个周期的过程中被固定在穿孔管。可以由这些固定管50 ML&#39;离心机&#39;的管，通过穿孔3毫米的孔，​​所有超过它的身体和将需要四个这种管子。 50毫升管可以方便的固定BALB / cJ小鼠，年龄超过8周。管可能需要适当大小的小鼠或一些不寻常的不同菌株，这是非常年轻的年龄超过50周的小鼠。 将固定管和孵化器内的一个笼子里保持在39.5°C。等待至少2个小时，所以笼子里，我mmobilization管的孵化器中达到的温度。 4。第一周期的热疗小鼠可能会​​遇到脱水在热疗曝光。因此，在实验开始之前，管理0.5 ml生理盐水腹腔内的每一个鼠标。使用26 G针1毫升注射器。 的笼子放置在孵化器中迅速取出，放置在每个固定管一个动物的笼子放置在孵化器背面。记录时间。关键是要保持笼子的孵化器每次在同一个地方。 90分钟后，取出笼从孵化器。松开固定管的动物，并把它们在笼子里，在常温下容易获得食物和水。将空的固定管的孵化器。 15分钟后，在第二个周期开始的热疗通过提供0。5 ml生理盐水腹腔内的每一个鼠标。 然后将它们固定在50毫升的穿孔固定管上，并放置在孵化器中。 5。随后的热疗周期对于第4个周期，它是必不可少的固定小鼠在热疗曝光期间的前两个周期，盐水注射热疗期间开始之前。 这些周期后，休息热疗完成后，小鼠获得适应环境并没有变得异常活跃的热疗过程中暴露。腹腔注射生理盐水可能没有必要，如果老鼠随手拿食物和水的休息时间。 6。完成的周期后，最后一步是检查动物不应该有任何的死亡在热疗周期。每种动物都有非常仔细地观看第一几个周期。快速查看任何润湿周围的吻。这种症状是脱水的签名。小鼠有此类症状，应立即给予0.5毫升生理盐水ip和其他在室温下。大多数的固定小鼠不需要任何额外的照顾，并会成功地承受压力的热疗。 完成所需的周期后，动物可能看起来虚弱和脱水，但他们会在几个小时内迅速恢复。 <img src="/files/ftp_upload/3801/3801fig1.jpg" alt="图1" /> 图1小鼠的培养箱中于39.5℃，当笼置于芯体温度变化笼90分钟后，从培养箱中取出，并放置在25℃。红色圆圈指示高的温度峰值。 <img src="/files/ftp_upload/3801/3801fig2.jpg" alt="图2" /> 芯体temperatu变化如图2所示。重新热疗周期期间的固定化的小鼠。固定动物后，高温峰值时不遵守笼被放置在39.5°C。

<ol>
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我们什么都没有透露。

<html>早在5000 BC，，埃及医生治疗肿瘤热。希腊人认识到，在一些医疗护理的热值。最古老的文本，摩西提到的"温泉（创世记36:24），，治疗提高身体的温度。 早期的技术使用气相微生物提取物诱导发热或热水浴诱导热疗1。然而，这些方法可以很容易地监视或控制。在我们的协议小鼠暴露于高热的控制和可重复的方式。 在此过程中，重要的是对?...

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protocol for long duration whole body hyperthermia in mice

热疗是一个通用术语，用来定义在核心体温高于正常的增加。它经常被用于描述增加芯体的温度的过程中观察到发热。热疗作为佐剂的使用已经出现作为一个有前途的癌症生物学领域中的肿瘤消退的过程。用于此目的，最重要的要求是具有可靠和均匀的加热协议。我们已经开发出了一个协议，用于热疗（全身）的小鼠。在这个协议中，动物被暴露于高温的周期为90分钟，然后通过15分钟的休息时间。在此期间，小鼠很容易获得食物和水。在最初的几个的热疗曝光周期的小鼠体温高尖峰，防止固定的动物。此外，施用生理盐水前几个周期中的脱水的影响减到最小。这个协议可以模拟条件下的小鼠12-24小时出现发烧一样。我们有使用8-12周龄BALB / Cj的雌性小鼠证明协议。

Watch this Scientific Journal Video about Protocol for Long Duration Whole Body Hyperthermia in Mice at JoVE.com

Protocol for Long Duration Whole Body Hyperthermia in Mice

本文介绍了一种协议，用于全身热疗的小鼠发烧一样，可以刺激条件下可达12-24小时。

持续时间较长的小鼠全身热疗协议

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

protocol-for-long-duration-whole-body-hyperthermia-in-mice

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

11.5K 次观看.  National Institute of Immunology. 本文介绍了一种协议，用于全身热疗的小鼠发烧一样，可以刺激条件下可达12-24小时。

视频: Protocol for Long Duration Whole Body Hyperthermia in Mice

Magnetic Resonance-Guided High Intensity Focused Ultrasound Generated Hyperthermia: A Feasible Treatment Method in a Murine Rhabdomyosarcoma Model

Magnetic resonance-guided high intensity focused ultrasound (MRgHIFU) is an established method for producing localized hyperthermia. Given the real-time imaging and acoustic energy modulation, this modality enables precise temperature control within a defined area. Many thermal applications are being explored with this noninvasive, nonionizing technology, such as hyperthermia generation, to release drugs from thermosensitive liposomal carriers. These drugs can include chemotherapies such as doxorubicin, for which targeted release is desired due to the dose-limiting systemic side effects, namely cardiotoxicity. Doxorubicin is a mainstay for treating a variety of malignant tumors and is commonly used in relapsed or recurrent rhabdomyosarcoma (RMS). RMS is the most common solid soft tissue extracranial tumor in children and young adults. Despite aggressive, multimodal therapy, RMS survival rates have remained the same for the past 30 years. To explore a solution for addressing this unmet need, an experimental protocol was developed to evaluate the release of thermosensitive liposomal doxorubicin (TLD) in an immunocompetent, syngeneic RMS mouse model using MRgHIFU as the source of hyperthermia for drug release.

Presented here is a protocol to use controlled hyperthermia, generated by magnetic resonance-guided high intensity focused ultrasound, to trigger drug release from temperature-sensitive liposomes in a rhabdomyosarcoma mouse model.

磁共振引导的高强度聚焦超声产生热疗:小鼠横纹肌肉瘤模型中的可行治疗方法

Magnetic resonance-guided high intensity focused ultrasound (MRgHIFU) is an established method for producing localized hyperthermia. Given the real-time ...

科研

癌症研究

Measuring Skeletal Muscle Thermogenesis in Mice and Rats

Skeletal muscle thermogenesis provides a potential avenue for better understanding metabolic homeostasis and the mechanisms underlying energy expenditure. Surprisingly little evidence is available to link the neural, myocellular, and molecular mechanisms of thermogenesis directly to measurable changes in muscle temperature. This paper describes a method in which temperature transponders are utilized to retrieve direct measurements of mouse and rat skeletal muscle temperature.
Remote transponders are surgically implanted within the muscle of mice and rats, and the animals are given time to recover. Mice and rats must then be repeatedly habituated to the testing environment and procedure. Changes in muscle temperature are measured in response to pharmacological or contextual stimuli in the home cage. Muscle temperature can also be measured during prescribed physical activity (i.e., treadmill walking at a constant speed) to factor out changes in activity as contributors to the changes in muscle temperature induced by these stimuli.
This method has been successfully used to elucidate mechanisms underlying muscle thermogenic control at the level of the brain, sympathetic nervous system, and skeletal muscle. Provided are demonstrations of this success using predator odor (PO; ferret odor) as a contextual stimulus and injections of oxytocin (Oxt) as a pharmacological stimulus, where predator odor induces muscle thermogenesis, and Oxt suppresses muscle temperature. Thus, these datasets display the efficacy of this method in detecting rapid changes in muscle temperature.

Mice and rats are surgically implanted with remote temperature transponders and then habituated to the testing environment and procedure. Changes in muscle temperature are measured in response to pharmacological or contextual stimuli in the home cage or during prescribed physical activity (i.e., treadmill walking at a constant speed).

测量小鼠和大鼠的骨骼肌产热

Skeletal muscle thermogenesis provides a potential avenue for better understanding metabolic homeostasis and the mechanisms underlying energy expenditure. ...

行为学

探索棕色脂肪组织中的代谢物交换

Arteriovenous Metabolomics to Measure In Vivo Metabolite Exchange in Brown Adipose Tissue

Brown adipose tissue (BAT) plays a crucial role in regulating metabolic homeostasis through a unique energy expenditure process known as non-shivering thermogenesis. To achieve this, BAT utilizes a diverse menu of circulating nutrients to support its high metabolic demand. Additionally, BAT secretes metabolite-derived bioactive factors that can serve as either metabolic fuels or signaling molecules, facilitating BAT-mediated intratissue and/or intertissue communication. This suggests that BAT actively participates in systemic metabolite exchange, an interesting feature that is beginning to be explored. Here, we introduce a protocol for in vivo mouse-level optimized BAT arteriovenous metabolomics. The protocol focuses on relevant methods for thermogenic stimulations and an arteriovenous blood sampling technique using Sulzer's vein, which selectively drains interscapular BAT-derived venous blood and systemic arterial blood. Next, a gas chromatography-based metabolomics protocol using those blood samples is demonstrated. The use of this technique should expand the understanding of BAT-regulated metabolite exchange at the inter-organ level by measuring the net uptake and release of metabolites by BAT.

作者聚焦:阐明脂肪组织代谢和疾病预防的新途径 

In this protocol, methods relevant for BAT-optimized arteriovenous metabolomics using GC-MS in a mouse model are outlined. These methods allow for the acquisition of valuable insights into BAT-mediated metabolite exchange at the organismal level.

动静脉代谢组学测量棕色脂肪组织 中的体内 代谢物交换

Brown adipose tissue (BAT) plays a crucial role in regulating metabolic homeostasis through a unique energy expenditure process known as non-shivering ...

生物化学

Mouse Body Temperature Measurement Using Infrared Thermometer During Passive Systemic Anaphylaxis and Food Allergy Evaluation

Mouse body temperature measurement is of paramount importance for investigating allergies and anaphylactic symptoms. Rectal probes for temperature readings is common, and they have been proven to be accurate and invaluable in this regard. However, this method of temperature measurement requires the mice to be anesthetized in order to insert the probe without injury to the animal. This limits the ability to observe other phenotypes of the mouse simultaneously. In order to investigate other phenotypes while measuring temperatures, rectal probes are not ideal, and another method is desired. Here, we introduce a noninvasive method of temperature measurement that foregoes the requirement for mouse anesthesia while maintaining equal reliability to rectal probes in measuring body temperature. We use an infrared thermometer that detects body surface temperatures at ranges between 2 and 150 mm. This method of body temperature measurement is successful in reliably replicating temperature change trends during passive system anaphylaxis experiments in mice. We show that body surface temperatures are about 2.0 &#176;C lower than rectal probe measurements, but the degree of temperature drop follows the same trend. Furthermore, we use the same technique to observe mice in a food allergy model to evaluate temperature and activity levels simultaneously.

Here we present a new method to accurately measure body temperature differences in passive systemic anaphylaxis (PSA) and food allergy mouse models using an infrared thermometer. This procedure has been accurately duplicated in previous PSA results.

红外测温仪在被动系统过敏反应和食品过敏评价中的应用

Mouse body temperature measurement is of paramount importance for investigating allergies and anaphylactic symptoms. Rectal probes for temperature ...

免疫与感染

Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has been challenged. Here, we fabricate iron oxide nanoparticle shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform. Iron oxide nanoparticles serve as both magnetic and photothermal agents. Once intravenously injected, NSMs can be magnetically guided to the tumor site. Ultrasound triggers the release of iron oxide nanoparticles, facilitating the penetration of nanoparticles deep into the tumor due to the cavitation effect of microbubbles. Thereafter, magnetic hyperthermia and photothermal therapy can be performed on the tumor for combinational cancer therapy, a solution for cancer resistance due to the tumor heterogeneity. In this protocol, the synthesis and characterization of NSMs including structural, chemical, magnetic and acoustic properties were performed. In addition, the anti-cancer efficacy by thermal therapy was investigated using in vitro cell cultures. The proposed delivery strategy and combination therapy holds great promise in cancer treatment to improve both delivery and anticancer efficacies.

Presented here is a protocol for the fabrication of iron oxide nanoparticle-shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform for magnetic hyperthermia and photothermal combination cancer therapy.

磁性、声学和光学三响应性微泡，用于磁性高温和波托托热组合癌症治疗

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has ...

生物工程

A Behavioral Screen for Heat-Induced Seizures in Mouse Models of Epilepsy

Transgenic mouse models have proved to be powerful tools in studying various aspects of human neurological disorders, including epilepsy. The SCN1A-associated genetic epilepsies comprise a wide spectrum of seizure disorders with incomplete penetrance and clinical variability. SCN1A mutations can result in a large variety of seizure phenotype ranging from simple, self-limited fever-associated febrile seizures (FS), moderate-level genetic epilepsy with febrile seizures plus (GEFS+) to more severe Dravet Syndrome (DS). Although FS are commonly seen in children below 6-7 years of age who do not have genetic epilepsy, FS in GEFS+ patients continue to occur into adulthood. Traditionally, experimental FS have been induced in mice by exposing the animal to a stream of dry air or heating lamps, and the rate of change in body temperature is often not well controlled. Here, we describe a custom-built heating chamber, with a plexiglass front, that is fitted with a digital temperature controller and a heater-equipped electric fan, which can send heated forced air into the test arena in a temperature-controlled manner. The body temperature of a mouse placed in the chamber, monitored through a rectal probe, can be increased to 40-42 &#176;C in a reproducible manner by increasing the temperature inside the chamber. Continual visual monitoring of the animals during the heating period demonstrates induction of heat-induced seizures in mice carrying an FS mutation at a body temperature that does not elicit behavioral seizures in wild-type litter mates. Animals can be easily removed from the chamber and placed on a cooling pad to rapidly return body temperature to normal. This method provides for a simple, rapid, and reproducible screening protocol for the occurrence of heat-induced seizures in epilepsy mouse models.

The goal of the method is to screen for hyperthermia or heat-induced seizures in mouse models. The protocol describes the use of a custom-built chamber with continuous monitoring of the body temperature to determine whether elevated body temperature leads to seizures.

癫痫小鼠模型中热诱发癫痫发作的行为筛查

Transgenic mouse models have proved to be powerful tools in studying various aspects of human neurological disorders, including epilepsy. The ...

神经科学

Short-Duration Hypothermia Induction in Rats using Models for Studies examining Clinical Relevance and Mechanisms

Therapeutic hypothermia (TH) is a powerful neuroprotective strategy that has provided robust evidence for neuroprotection in pre-clinical studies of neurological disorders. Despite strong pre-clinical evidence, TH has not shown efficacy in clinical trials of most neurological disorders. The only successful trials employing therapeutic hypothermia were related to cardiac arrest in adults and hypoxic ischemic injury in neonates. Further investigations into the parameters of its use, and study design comparisons between pre-clinical and clinical studies, are warranted. This article demonstrates two methods of short-duration hypothermia induction. The first method allows for rapid hypothermia induction in rats using ethanol spray and fans. This method works by cooling the skin, which has been less commonly used in clinical trials and may have different physiological effects. Cooling is much more rapid with this technique than is achievable in human patients due to differences in surface area to volume ratio. Along with this, a second method is also presented, which allows for a clinically achievable cooling rate for short-duration hypothermia. This method is easy to implement, reproducible and does not require active skin cooling.

This article describes two methods of whole-body short-duration hypothermia induction in rats. The first, rapid induction method, employs active cooling using fans and ethanol spray for a rapid decrease in temperature. The second method is a gradual cooling method. This is achieved using the combination of isoflurane anesthesia and the reduction of temperature settings on the homeothermic heat mat. This results in a gradual decrease in core body temperature without the use of any external cooling devices.&#160;

使用研究临床相关性和机制的模型在大鼠中短时间体温过低诱导

Therapeutic hypothermia (TH) is a powerful neuroprotective strategy that has provided robust evidence for neuroprotection in pre-clinical studies of ...

医学

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

小鼠劳力性中暑的临床前模型

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

持续时间较长的小鼠全身热疗协议

摘要

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磁共振引导的高强度聚焦超声产生热疗:小鼠横纹肌肉瘤模型中的可行治疗方法

测量小鼠和大鼠的骨骼肌产热

动静脉代谢组学测量棕色脂肪组织中的体内代谢物交换

动静脉代谢组学测量棕色脂肪组织中的体内代谢物交换

动静脉代谢组学测量棕色脂肪组织中的体内代谢物交换

红外测温仪在被动系统过敏反应和食品过敏评价中的应用

磁性、声学和光学三响应性微泡，用于磁性高温和波托托热组合癌症治疗

癫痫小鼠模型中热诱发癫痫发作的行为筛查

使用研究临床相关性和机制的模型在大鼠中短时间体温过低诱导

小鼠劳力性中暑的临床前模型

持续时间较长的小鼠全身热疗协议

摘要

探索更多视频

此视频中的章节

磁共振引导的高强度聚焦超声产生热疗:小鼠横纹肌肉瘤模型中的可行治疗方法

测量小鼠和大鼠的骨骼肌产热

动静脉代谢组学测量棕色脂肪组织 中的体内 代谢物交换

动静脉代谢组学测量棕色脂肪组织 中的体内 代谢物交换

动静脉代谢组学测量棕色脂肪组织 中的体内 代谢物交换

红外测温仪在被动系统过敏反应和食品过敏评价中的应用

磁性、声学和光学三响应性微泡，用于磁性高温和波托托热组合癌症治疗

癫痫小鼠模型中热诱发癫痫发作的行为筛查

使用研究临床相关性和机制的模型在大鼠中短时间体温过低诱导

小鼠劳力性中暑的临床前模型

动静脉代谢组学测量棕色脂肪组织中的体内代谢物交换

动静脉代谢组学测量棕色脂肪组织中的体内代谢物交换

动静脉代谢组学测量棕色脂肪组织中的体内代谢物交换