<ol style="padding:5px;"><li> 설정 장비 <ol style="padding:5px;"><li> 실내 온도가 ° C.에서 20 위에 실험실에서 영역을 선택 </li><li> 에어컨 통풍구 및 / 또는 큰 소음이 근처에 위치를 피하십시오. </li><li> 같은 향수, 향수, 화학, 그리고 클리너로 자극 및 / 또는 스트레스의 원인이 될 수 있습니다 악취로 동물을 노출하지 마십시오. </li><li> 코다 컨트롤러를 켭니다. </li><li> 온난화 플랫폼의 전원을 켜고 레벨 3으로 설정합니다. 실내 온도가 25 이상이면 ° C는 레벨 1 또는 2에서 시작합니다. </li><li> 컨트롤러 진단 테스트 <ol style="padding:5px;"><li> 코다 소프트웨어를 엽니다. </li><li> 그것을 클릭하여 코다 장치를 선택합니다. </li><li> &quot;테스트 선택한 장치&quot;를 클릭하십시오. </li><li> &quot;테스트&quot;를 테스트하고를 클릭 수갑 및 채널을 선택합니다. </li><li> 테스트가 완료되면, 장치 테스트 창을 닫습니다. </li></ol></li></ol></li><li> 설정 소프트웨어 <ol style="padding:5px;"><li> 인사 및 동물 관리 <ol style="padding:5px;"><li> 인사 관리 <ol style="padding:5px;"><li> 선택 도구&gt; 인사 관리. </li><li> 이름을 변경하려면 &quot;연구원 1&quot;을 클릭하십시오. </li><li> 추가 연구를 추가하는 후속 행을 클릭합니다. </li><li> 기술자를 수정하거나 추가하려면 &quot;기술자 탭&quot;을 클릭합니다. </li><li> 쥐가 드나 마우스, 쉽게 식별이 동물의 이름과 동물의 유형을 선택하려면 &quot;표본 탭&quot;을 클릭합니다. </li><li> 동물 종류와 감도를 변경하려면 &quot;동물 탭&quot;을 클릭합니다. </li></ol></li></ol></li><li> 새 실험 <ol style="padding:5px;"><li> 파일&gt; 새로 작성&gt; 실험을 선택 </li><li> 새 실험을 시작 <ol style="padding:5px;"><li> 실험 이름을 입력합니다. </li><li> 주요 연구원을 선택합니다. </li><li> 시작 날짜를 선택하고 &quot;다음&quot;을 클릭합니다. </li></ol></li><li> 기본 세션 정보 <ol style="padding:5px;"><li> 세션 이름을 입력 </li><li> 5 &quot;Acclimation주기&quot;를 설정합니다. 당신은 0으로 설정 acclimation 사이클을 포함하지 않도록 설정하는 경우. , 홀더와 수갑으로 동물을 acclimating 도와 실험에 추가 사이클을 추가 5 연속 일 동안 실험을 실행합니다. </li><li> 1 &quot;설정 번호&quot;를 설정합니다. </li><li> 여러 집합에 대한 30 초 &quot;설정 사이의 시간&quot;을 설정 </li><li> 20주기 위해 &quot;설정 당 사이클&quot;을 설정합니다. </li><li> 오초에 &quot;사이클 사이의 시간&quot;을 설정하고 &quot;다음&quot;을 클릭합니다. </li></ol></li></ol></li><li> 표본 선택 <ol style="padding:5px;"><li style="list-style-type:none;"><ol style="padding:5px;"><li> 표본 수영장에서 표본을 선택하고 사용 가능한 첫 번째 채널을 지정하고 &quot;다음&quot;을 클릭하십시오. </li></ol></li><li> 세션 매개 변수 <ol style="padding:5px;"><li> 250mmHg에 &quot;최대 폐색 압력&quot;을 설정합니다. </li><li> 마우스, 쥐 때까지 15 초 20 초 &#39;디플레이션 시간 &quot;을 설정합니다. </li><li> 15ul에 &quot;최소 볼륨&quot;을 설정합니다. </li><li> 그래프 당 하나의 채널에 &quot;표시 스타일&quot;을 설정합니다. </li><li> 추가 연구자와 기술자를 추가하고 &quot;다음&quot;을 클릭하십시오. </li></ol></li><li> 세션 스크립트 <ol style="padding:5px;"><li> 세션 스크립트를 검토하고 &quot;다음&quot;을 클릭하십시오. </li><li> 당신은 동물 준비 섹션을 완료할 때까지 마침을 선택하지 마십시오. </li></ol></li></ol></li></ol></li><li> 동물 준비 <ol style="padding:5px;"><li> 설정 동물 <ol style="padding:5px;"><li> 동물 홀더 <ol style="padding:5px;"><li> 기차와 동물 acclimation을 향상 시키며, 실험에 추가 사이클을 추가하고 다섯 경기를 연속 일 동안 실험을 실행합니다. 당신은 또한 실제 공부하기 전에 연속 3 일간 15 분 홀더에 동물을 배치하여 acclimation와 도움을 줄 수 있습니다. </li><li> 꼬리로 동물을 데리러 부드럽게 코 콘의 열린 끝을 얼굴 홀더의 뒷면에 동물을 배치하여 보유자의 각 동물을 놓으십시오. 참고 : 선택한 홀더의 크기와 관련하여 동물의 크기. 동물 자유롭게 홀더를 입력하실 수 있습니다. </li><li> 조심스럽게 뒷문에있는 빨간 나사 뚜껑을 설정하여 소유자에게 뒷문을 확보. 주의가 뒷문을 확보하면서 꼬리 또는 기타 신체 부분을 곤란하게하지 않도록 주의해야한다. </li><li> 동물의 움직임을 제한, 뒷문쪽으로 코 콘 슬라이드. 코 콘은 홀더 내부 동안 주변을 공전의 동물을 제한하는 위치에 있어야합니다. </li><li> 지정된 위치에 온난 화 플랫폼에 홀더를 놓으십시오. 일정 기간 동안 동물의 동일한 그룹을 테스트하는 경우 BP 측정의 일관성 및 정확도를 향상시키기 위해 온난 화 플랫폼 홀더의 위치를​​ 회전해야합니다. 적어도 5 분 자에 적응하기 위해 동물을 허용합니다. </li><li> 동물을 만지거나 처리하지 마십시오 자 안에있는 동안. 증가 연락처는 동물을 짜증나게 수 있습니다. </li><li> 무인 홀더에있는 동물을 두지 마십시오. </li></ol></li><li> 커프 위치 <ol s타일​​은 = &quot;패딩 : 5px;&quot;&gt; <li> 를 통해 스레드 꼬리 &quot;를 폐색 커프가.&quot;강제하지 않고 가능한 한 꼬리의 기지에 가까운이 수갑 플레이스. </li><li> 홀더의 맨 뒤쪽에있는 노치에있는 &quot;폐색의 커프 튜브&quot;를 고정합니다. </li><li> &quot;폐색 커프 &#39;의 2mm 이내에 그것을 배치,&quot;VPR 커프&#39;를 통해 꼬리를 스레드. </li><li> 홀더의 맨 뒤쪽에있는 노치에있는 &quot;VPR의 커프 튜브&quot;를 고정합니다. </li><li> 코다 컨트롤러에 수갑을 연결합니다. </li></ol></li><li> 동물 온도 <ol style="padding:5px;"><li> 적어도 5 분 thermoregulate에 동물을 허용합니다. </li><li> 적외선 온도계와 함께 자주 동물의 온도를 기록합니다. 동물 온도 32 35 ° C.되어야합니다 </li></ol></li></ol></li></ol></li><li> 실험 <ol style="padding:5px;"><li> 실험 시작 <ol style="padding:5px;"><li> &quot;Finish&quot;를 클릭하고 실험 즉시 시작됩니다. </li></ol></li><li> 최종 실험 <ol style="padding:5px;"><li> 동물 <ol style="padding:5px;"><li> 즉시 수갑과 홀더에서 동물을 제거합니다. </li><li> &quot;간단한 세션 요약 보고서&quot;가 표시되면 해당 창구로 동물을 반환합니다. </li></ol></li><li> 데이터 수집 <ol style="padding:5px;"><li> 허용주기가 자동으로 코다 응용 프로그램 내에서 스프레드 시트 형식으로 표시됩니다 마지막주기의 완료 후, &quot;Excel로 내보내기&quot;버튼을 클릭하십시오. </li><li> 에서 파일을 폴더를 선택하고 파일 이름을 지정하고 &quot;저장&quot;을 클릭하십시오. </li><li> 엑셀 응용 프로그램을 열고 &quot;열기&quot;를 클릭하십시오. </li><li> &quot;. csv 파일 저장&quot;을 표시하는 모든 파일 형식을보기 파일을 클릭합니다. </li></ol></li><li> 데이터 처리 <ol style="padding:5px;"><li> 엑셀 파일 내의 데이터를 처리하는 방법에는 여러 가지가 있습니다. 일반적인 연습은 평균 및 표준 편차를 얻을 수 있습니다. </li><li> 표준 편차가 30보다 큰 경우에는 측정을 삭제합니다. </li><li> 또한, 거부 사이클을 볼 수 있으며 전체 데이터베이스를 쉽게 Excel로 내보낼 수 있습니다. 자세한 내용은 사용자 설명서를 참조하십시오. </li></ol></li></ol></li></ol></li></ol>

The authors have nothing to disclose.

measuring blood pressure in mice using volume pressure recording, a tail-cuff method

코다 8 채널 높은 처리량 비침습 혈압 시스템은 동시에 8 쥐 또는 쥐에 혈압을 측정합니다. 코다 꼬리 즉석 시스템은 꼬리 혈액 볼륨을 결정하여 혈압을 측정하는 볼륨 압력 녹음 (VPR)를 사용합니다. 개별 펄스 신호를 얻기 위해 필요없이 특별히 고안된 차동 압력 변환기 및 폐색 꼬리 즉석 측정 꼬리의 총 혈액 볼륨. 특별한주의는 가장 정확한 혈압 수치를 도출하기 위해 폐색의 즉석의 길이에 여유입니다. VPR 쉽게 같은 C57BL6 생쥐로 어두운 피부 설치류에 판독을 구하고 호환 MRI입니다 수 있습니다. 코다 시스템은 여섯 (6) 다른 혈압 파라미터 측정을 제공, 수축기 및 diastolic 혈압, 심장 박동은, 혈압, 꼬리 혈액 흐름, 그리고 꼬리 혈액 볼륨을 의미합니다. 성능은 깨어 또는 anesthetized 마우스 또는 쥐 중 하나를 만들 수 있습니다. 코다 시스템은 컨트롤러, 노트북 컴퓨터, 소프트웨어, 수갑, 동물 소유자, 적외선 온난 패드, 그리고 적외선 온도계를 포함합니다. 900g 크기로 쥐에 8g만큼 작은 생쥐를위한 7 개의 홀더 크기가 있습니다.

Watch this Scientific Journal Video about Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method at JoVE.com

Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

코다 8 채널 높은 처리량 비침습 혈압 시스템은 동시에 8 쥐 또는 쥐에 혈압을 측정합니다. 이 꼬리 즉석 시스템은 꼬리 혈액 볼륨을 결정하여 혈압을 측정하는 볼륨 압력 녹음 (VPR)를 사용합니다.

볼륨 압력 녹음, 꼬리 즉석 방법을 사용하여 마우스의 측정 혈압

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA ...

measuring-blood-pressure-mice-using-volume-pressure-recording-tail

Research

JoVE Journal

Biology

51.3K 조회수.  University of Kentucky. 코다 8 채널 높은 처리량 비침습 혈압 시스템은 동시에 8 쥐 또는 쥐에 혈압을 측정합니다. 이 꼬리 즉석 시스템은 꼬리 혈액 볼륨을 결정하여 혈압을 측정하는 볼륨 압력 녹음 (VPR)를 사용합니다.

비디오: Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

Pressure-polishing Pipettes for Improved Patch-clamp Recording

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording. We first developed this method for fabricating pipettes suitable for recording from small (&lt;3 m) neuronal cell bodies. The basic principal is similar to glass-blowing and combines air pressure and heat to modify the shape of patch pipettes prepared by a conventional micropipette puller. It can be applied to so-called soft (soda lime) and hard (borosilicate) glasses. Generally speaking, pressure polishing can reduce pipette resistance by 25% without decreasing the diameter of the tip opening (Goodman and Lockery, 2000). It can be applied to virtually any type of glass and requires only the addition of a high-pressure valve and fitting to a microforge. This technique is essential for recording from ultrasmall cells (&lt;5 m) and can also improve single-channel recording by minimizing pipette resistance. The blunt shape is also useful for perforated-patch clamp recording since this tip shape results in a larger membrane bleb available for perforation.

This is a guide to modifying the shape of glass micropipettes. Specifically, by using heat and air pressure the taper is widened without increasing the tip opening, leading to lower pipette resistance. This is critical to obtain low noise recordings of small cells but is useful in many applications.

향상된 패치 - 클램프 녹음 압력 연마 Pipettes

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

연구

생물학

Making Gynogenetic Diploid Zebrafish by Early Pressure

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow F1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. Streisinger et al.1,2 described methods for making gynogenetic (homozygous) diploid zebrafish by activating zebrafish eggs with ultraviolet light-inactivated sperm and preventing either the second meiotic or the first zygotic cell division using physical treatments (heat or pressure) that deploymerize microtubules. The "early pressure" (EP) method blocks the meiosis II, which occurs shortly after fertilization. The EP method produces a high percentage of viable embryos that can develop to fertile adults of either sex. The method generates embryos that are homozygous at all loci except those that were separated from their centromere by recombination during meiosis I. Homozygous mutations are detected in EP clutches at between 50% for centromeric loci and less than 1% for telomeric loci. This method is reproduced verbatim from the Zebrafish Book3.

This is a method for generating gynogenetic diploid zebrafish embryos (embryos whose only genetic contribution comes from the mother) by blocking the second meiotic division immediately after fertilization with ultraviolet light-inactivated sperm. EP embryos are not fully homozygous due to recombination during the first meiotic division, however they are homozygous at all loci that have not been separated from their centromere by recombination.

초기 압력에 의해 Gynogenetic의 Diploid의 Zebrafish 만들기

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from ...

Ambulatory ECG Recording in Mice

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although the surface ECG is utilized for short-term measurements of waveform intervals, it is not practical for long-term studies of heart rate variability or the capture of rare episodes of arrhythmias. Implantable ECG telemeters offer the advantages of simple surgical implantation, long-term recording of electrograms in ambulatory mice, and scalability with simultaneous recordings of multiple animals. Here, we present a step-by-step guide to the implantation of telemeters for ambulatory ECG recording in mice. Careful adherence to aseptic technique is required for favorable survival results with the possibility of implantation and recording from weeks to months. Thus, implantable ECG telemetry is a valuable tool for detection of critical information on cardiac electrophysiology in ambulatory animal models such as the mouse. 

Telemetric ECG has emerged as an essential tool in evaluating animal models for cardiac arrhythmias and sudden cardiac death. Here, we present a stepwise guide to telemetric ECG recordings for application in long-term ambulatory ECG monitoring in mice.

마우스의 외래 ECG 기록

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although ...

Isolation and Culture of Pulmonary Endothelial Cells from Neonatal Mice

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells (HUVECs) have shown to be convenient, easy to obtain and culture, and thus are the most widely studied endothelial cells. However, for research focused on processes like angiogenesis, permeability or many others, microvascular endothelial cells (ECs) are a much more physiologically relevant model to study 2. Furthermore, ECs isolated from knockout mice provide a useful tool for analysis of protein function ex vivo. Several approaches to isolate and culture microvascular ECs of different origin have been reported to date 3-7, but consistent isolation and culture of pure ECs is still a major technical problem in many laboratories. Here, we provide a step-by-step protocol on a reliable and relatively simple method of isolating and culturing mouse lung endothelial cells (MLECs). In this approach, lung tissue obtained from 6- to 8-day old pups is first cut into pieces, digested with collagenase/dispase (C/D) solution and dispersed mechanically into single-cell suspension. MLECS are purified from cell suspension using positive selection with anti-PECAM-1 antibody conjugated to Dynabeads using a Magnetic Particle Concentrator (MPC). Such purified cells are cultured on gelatin-coated tissue culture (TC) dishes until they become confluent. At that point, cells are further purified using Dynabeads coupled to anti-ICAM-2 antibody. MLECs obtained with this protocol exhibit a cobblestone phenotype, as visualized by phase-contrast light microscopy, and their endothelial phenotype has been confirmed using FACS analysis with anti-VE-cadherin 8 and anti-VEGFR2 9 antibodies and immunofluorescent staining of VE-cadherin. In our hands, this two-step isolation procedure consistently and reliably yields a pure population of MLECs, which can be further cultured. This method will enable researchers to take advantage of the growing number of knockout and transgenic mice to directly correlate in vivo studies with results of in vitro experiments performed on isolated MLECs and thus help to reveal molecular mechanisms of vascular phenotypes observed in vivo.

Here, we describe a protocol for isolation and culture of murine pulmonary endothelial cells. This method comprises mechanic and enzymatic lung tissue dissociation as well as a 2-step purification process using anti-PECAM-1 and anti-ICAM-2 antibodies conjugated to magnetic beads, which produces a pure endothelial cell population of mostly microvascular origin.

신생아 마우스의 폐 내피 세포의 분리 및 문화

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

Assessing Murine Resistance Artery Function Using Pressure Myography

Pressure myograph systems are exquisitely useful in the functional assessment of small arteries, pressurized to a suitable transmural pressure. The near physiological condition achieved in pressure myography permits in-depth characterization of intrinsic responses to pharmacological and physiological stimuli, which can be extrapolated to the in vivo behavior of the vascular bed. Pressure myograph has several advantages over conventional wire myographs. For example, smaller resistance vessels can be studied at tightly controlled and physiologically relevant intraluminal pressures. Here, we study the ability of 3rd order mesenteric arteries (3-4 mm long), preconstricted with phenylephrine, to vaso-relax in response to acetylcholine. Mesenteric arteries are mounted on two cannulas connected to a pressurized and sealed system that is maintained at constant pressure of 60 mmHg. The lumen and outer diameter of the vessel are continuously recorded using a video camera, allowing real time quantification of the vasoconstriction and vasorelaxation in response to phenylephrine and acetylcholine, respectively.
	To demonstrate the applicability of pressure myography to study the etiology of cardiovascular disease, we assessed endothelium-dependent vascular function in a murine model of systemic hypertension. Mice deficient in the &alpha;1 subunit of soluble guanylate cyclase (sGC&alpha;1-/-) are hypertensive when on a 129S6 (S6) background (sGC&alpha;1-/-S6) but not when on a C57BL/6 (B6) background (sGC&alpha;1-/-B6). Using pressure myography, we demonstrate that sGC&alpha;1-deficiency results in impaired endothelium-dependent vasorelaxation. The vascular dysfunction is more pronounced in sGC&alpha;1-/-S6 than in sGC&alpha;1-/-B6 mice, likely contributing to the higher blood pressure in sGC&alpha;1-/-S6 than in sGC&alpha;1-/-B6 mice. 
	Pressure myography is a relatively simple, but sensitive and mechanistically useful technique that can be used to assess the effect of various stimuli on vascular contraction and relaxation, thereby augmenting our insight into the mechanisms underlying cardiovascular disease.

In pressure myography, an intact small segment of a vessel is mounted onto two small cannulas and pressurized to a suitable luminal pressure. Here, we describe the method to measure vasorelaxation response of the mouse 3rd order mesenteric arteries in c57 and sGC&alpha;1-/- mice using pressure myography.

압력 Myography를 사용 쥐과 저항 동맥의 기능을 평가

Pressure myograph systems are  exquisitely useful in the functional assessment of small arteries, pressurized  to a suitable transmural pressure. The  ...

One-channel Cell-attached Patch-clamp Recording

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate depends on properties intrinsic to each channel protein as well as the mechanism by which it is generated and controlled and represents an important area of current research. Information about the operational dynamics of ion channel proteins can be obtained by observing long stretches of current produced by a single molecule. Described here is a protocol for obtaining one-channel cell-attached patch-clamp current recordings for a ligand gated ion channel, the NMDA receptor, expressed heterologously in HEK293 cells or natively in cortical neurons. Also provided are instructions on how to adapt the method to other ion channels of interest by presenting the example of the mechano-sensitive channel PIEZO1. This method can provide data regarding the channel&#8217;s conductance properties and the temporal sequence of open-closed conformations that make up the channel&#8217;s activation mechanism, thus helping to understand their functions in health and disease.

Described here is a procedure for obtaining long stretches of current recording from one ion channel with the cell-attached patch-clamp technique. This method allows for observing, in real time, the pattern of open-close channel conformations that underlie the biological signal. These data inform about channel properties in undisturbed biological membranes.

1 채널의 세포 부착 된 패치 클램프 녹음

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative assessment of lung function in rodent models is an important tool in the development of future therapies. Commonly used techniques for assessing respiratory function including invasive plethysmography and forced oscillation. While these techniques provide valuable information, data collection can be fraught with artefacts and experimental variability due to the need for anesthesia and/or invasive instrumentation of the animal. In contrast, unrestrained whole-body plethysmography (UWBP) offers a precise, non-invasive, quantitative way by which to analyze respiratory parameters. This technique avoids the use of anesthesia and restraints, which is common to traditional plethysmography techniques. This video will demonstrate the UWBP procedure including the equipment set up, calibration and lung function recording. It will explain how to analyze the collected data, as well as identify experimental outliers and artefacts that results from animal movement. The respiratory parameters obtained using this technique include tidal volume, minute volume, inspiratory duty cycle, inspiratory flow rate and the ratio of inspiration time to expiration time. UWBP does not rely on specialized skills and is inexpensive to perform. A key feature of UWBP, and most appealing to potential users, is the ability to perform repeated measures of lung function on the same animal.

The assessment of respiratory physiology has traditionally relied upon techniques, which require restraint or sedation of the animal. Unrestrained whole-body plethysmography, however, provides precise, non-invasive, quantitative analysis of respiratory physiology in animal models. In addition, the technique allows repeated respiratory assessment of mice allowing for longitudinal studies.

무제한 전신 혈량 측정법을 사용하여 마우스의 호흡 기능을 측정

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

A Method for Screening and Validation of Resistant Mutations Against Kinase Inhibitors

The discovery of BCR/ABL as a driver oncogene in chronic myeloid leukemia (CML) resulted in the development of Imatinib, which, in fact, demonstrated the potential of targeting the kinase in cancers by effectively treating the CML patients. This observation revolutionized drug development to target the oncogenic kinases implicated in various other malignancies, such as, EGFR, B-RAF, KIT and PDGFRs. However, one major drawback of anti-kinase therapies is the emergence of drug resistance mutations rendering the target to have reduced or lost affinity for the drug. Understanding the mechanisms employed by resistant variants not only helps in developing the next generation inhibitors but also gives impetus to clinical management using personalized medicine. We reported a retroviral vector based screening strategy to identify the spectrum of resistance conferring mutations in BCR/ABL, which has helped in developing the next generation BCR/ABL inhibitors. Using Ruxolitinib and JAK2 as a drug target pair, here we describe in vitro screening methods that utilizes the mouse BAF3 cells expressing the random mutation library of JAK2 kinase.

Emergence of genetic resistance against kinase inhibitor therapy poses significant challenge for effective cancer therapy. Identification and characterization of resistant mutations against a newly developed drug helps in better clinical management and next generation drug design. Here, we describe our protocol for in vitro screening and validation of resistant mutations.

키나제 억제제에 대해 내성 돌연변이의 검사 및 검증을위한 방법

The discovery of BCR/ABL as a driver oncogene in chronic myeloid leukemia (CML) resulted in the development of Imatinib, which, in fact, demonstrated the ...

Cardiac Pressure-Volume Loop Analysis Using Conductance Catheters in Mice

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular systolic and diastolic function. Measures of ventricular contractility and compliance are obtained through examination of cardiac response to changes in afterload and preload. These techniques were originally developed nearly three decades ago to measure cardiac function in large mammals and humans. The application of these analyses to small mammals, such as mice, has been accomplished through the optimization of microsurgical techniques and creation of conductance catheters. Conductance catheters allow for estimation of the blood pool by exploiting the relationship between electrical conductance and volume. When properly performed, these techniques allow for testing of cardiac function in genetic mutant mouse models or in drug treatment studies. The accuracy and precision of these studies are dependent on careful attention to the calibration of instruments, systematic conduct of hemodynamic measurements and data analyses. We will review the methods of conducting pressure-volume loop experiments using a conductance catheter in mice.

Cardiac pressure-volume loop analysis is the most comprehensive way to measure cardiac function in the intact heart. We describe a technique to perform and analyze cardiac pressure volume loops, using conductance catheters.

마우스에서 컨덕턴스 카테터를 사용하여 심장 압력 볼륨 루프 분석

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular ...

Measuring Pressure Volume Loops in the Mouse

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the mouse provides great potential to explore cardiac function at the molecular level. The mouse&#39;s small size does present some challenges in regards to performing detailed cardiac phenotyping. Miniaturization and other advancements in technology have made many methods of cardiac assessment possible in the mouse. Of these, the simultaneous collection of pressure and volume data provides a detailed picture of cardiac function that is not available through any other modality. Here a detailed procedure for the collection of pressure-volume loop data is described. Included is a discussion of the principles underlying the measurements and the potential sources of error. Anesthetic management and surgical approaches are discussed in great detail as they are both critical to obtaining high quality hemodynamic measurements. The principles of hemodynamic protocol development and relevant aspects of data analysis are also addressed.

This manuscript describes a detailed protocol for the collection of pressure-volume data from the mouse.

마우스에 압력 볼륨 루프 측정

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the ...