Jove-Journal of Visualized Experiments最新文献

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Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model.
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-29 DOI: 10.3791/67189
Yanjiani Xu, Jialiang Zhang, Jing Zhou, Yaoyu Zhang, Fangyang Huang, Mao Chen
{"title":"Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model.","authors":"Yanjiani Xu, Jialiang Zhang, Jing Zhou, Yaoyu Zhang, Fangyang Huang, Mao Chen","doi":"10.3791/67189","DOIUrl":"https://doi.org/10.3791/67189","url":null,"abstract":"<p><p>The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus (T2DM), remain poorly understood. The myocardial toxicity associated with T2DM is attributed to factors such as lipotoxicity, glucotoxicity, oxidative stress, reduced cardiac efficiency, and lipoapoptosis. Compared to rats, mice offer greater accessibility, cost-effectiveness, and broader applicability for animal experiments. Insulin resistance and impaired insulin secretion are crucial factors in the pathophysiology of T2DM. We introduce a novel nongenetic murine model that replicates the progression of human DbCM induced by a combination of high-fat diet (HFD) feeding and streptozotocin (STZ) injection. In this study, we used wild-type C57BL/6J mice, administering an HFD regimen for 12 weeks, followed by intraperitoneal injections of STZ for an additional 12 weeks to induce characteristic manifestations of T2DM. We conducted oral glucose tolerance tests and measured serum insulin concentrations to confirm the development of insulin resistance and insufficient insulin secretion. Cardiac structure and function were rigorously assessed through noninvasive transthoracic echocardiography. Pathological characteristics were evaluated through Masson's trichrome staining and wheat germ agglutinin (WGA) staining, revealing pathological features related to DbCM. Therefore, we provide a robust and versatile method for establishing a nongenetic murine model of DbCM.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Electroporation of Sliced Human Cortical Organoids for Studies of Gene Function. 电穿孔切片人皮质器官组织用于基因功能研究
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-29 DOI: 10.3791/67598
Annika Kolodziejczyk, Janine Hoffmann, Paula Cubillos, Mareike Albert
{"title":"Electroporation of Sliced Human Cortical Organoids for Studies of Gene Function.","authors":"Annika Kolodziejczyk, Janine Hoffmann, Paula Cubillos, Mareike Albert","doi":"10.3791/67598","DOIUrl":"https://doi.org/10.3791/67598","url":null,"abstract":"<p><p>Human cortical organoids have become important tools for studying human brain development, neurodevelopmental disorders, and human brain evolution. Studies analyzing gene function by overexpression or knockout have been instrumental in animal models to provide mechanistic insights into the regulation of neocortex development. Here, we present a detailed protocol for CRISPR/Cas9-mediated acute gene knockout by electroporation of sliced human cortical organoids. The slicing of cortical organoids aids the identification of ventricle-like structures for injection and subsequent electroporation, making this a particularly well-suited model for acute genetic manipulation during human cortical development. We describe the design of guide RNAs and the validation of targeting efficiency in vitro and in cortical organoids. Electroporation of cortical organoids is performed at mid-neurogenic stages, enabling the targeting of most major cell classes in the developing neocortex, including apical radial glia, basal progenitor cells, and neurons. Taken together, the electroporation of sliced human cortical organoids represents a powerful technique to investigate gene function, gene regulation, and cell morphology during cortical development.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Orienteering as a Tool for Cognitive Research: An Implementation Guide. 作为认知研究工具的定向运动:实施指南》。
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-29 DOI: 10.3791/66833
Emma E Waddington, Jennifer J Heisz
{"title":"Orienteering as a Tool for Cognitive Research: An Implementation Guide.","authors":"Emma E Waddington, Jennifer J Heisz","doi":"10.3791/66833","DOIUrl":"https://doi.org/10.3791/66833","url":null,"abstract":"<p><p>The sport of orienteering combines physical activity with spatial navigation. Using only a map and a compass, the orienteer must locate a series of checkpoints over unfamiliar terrain using any navigational route they choose and while moving as quickly as possible. Although expert orienteers have superior spatial memory and navigational abilities, even a single session of orienteering can benefit cognition, suggesting that orienteering may be a promising way to train the brain. Research interventions involving orienteering may be especially beneficial for staving off Alzheimer's disease and related dementias that are afflicted by early impairments in wayfinding and spatial cognition. Though orienteering has gained traction in recent literature, certain barriers exist for researchers who are unfamiliar with the sport and wish to implement an intervention. Specifically, a lack of research-based resources for creating orienteering maps and courses may prevent those wishing to study orienteering from designing an intervention. Therefore, this report provides the fundamental information needed to develop orienteering maps and courses and how to implement orienteering interventions in a research setting.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822874","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Identification and Quantification of Deranged Metabolites in Critically Ill Patients Using NMR-Based Metabolomics. 利用基于核磁共振的代谢组学鉴定和定量重症患者体内紊乱的代谢物。
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-29 DOI: 10.3791/67319
Anamika Singh, Mohd Adnan Siddiqui, Afzal Azim, Neeraj Sinha
{"title":"Identification and Quantification of Deranged Metabolites in Critically Ill Patients Using NMR-Based Metabolomics.","authors":"Anamika Singh, Mohd Adnan Siddiqui, Afzal Azim, Neeraj Sinha","doi":"10.3791/67319","DOIUrl":"https://doi.org/10.3791/67319","url":null,"abstract":"<p><p>Metabolomics is emerging as a significant approach to reflect the individual's response to pathophysiological conditions. Nuclear magnetic resonance (NMR) spectroscopy has evolved as a tool to identify metabolic dysregulations in critically ill patients afflicted with conditions like acute respiratory distress syndrome (ARDS), severe acute pancreatitis (SAP), acute kidney injury (AKI), and sepsis. The spectral data from the serum sample of the study and control group are recorded using an 800 MHz NMR spectrometer and processed using NMR processing and analysis tools. Furthermore, a rigorous statistical analysis, such as univariate and multivariate tests, is performed to pinpoint significant metabolites, which are then accurately identified and quantified using NMR metabolite quantification software. Additionally, pathway analysis highlights the deranged biochemical cycles that result in the severity of illness. Through this comprehensive approach, researchers aim to gain deeper insights into the metabolic alterations associated with these critical illnesses, potentially paving the way for a better understanding of the disease and improved diagnostics and treatment strategies.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A Trap-Vaccinate-Release Protocol for Immunization of Skunks and Additional Rabies Vectors Against Rabies. 对臭鼬和其他狂犬病病媒进行狂犬病免疫接种的诱捕-疫苗接种-释放方案。
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-29 DOI: 10.3791/65117
Lias Hastings, David L Bergman
{"title":"A Trap-Vaccinate-Release Protocol for Immunization of Skunks and Additional Rabies Vectors Against Rabies.","authors":"Lias Hastings, David L Bergman","doi":"10.3791/65117","DOIUrl":"https://doi.org/10.3791/65117","url":null,"abstract":"<p><p>Oral vaccination of wildlife against rabies via the distribution of vaccine-laden baits is used widely as a management tool in Europe and North America. Over the past several decades, successful programs have targeted important reservoirs, including coyotes, foxes, raccoon dogs, and raccoons, for prevention and control. However, other species (e.g., skunks) or certain habitats (e.g., urban) may be less amenable to oral vaccination, requiring consideration of additional strategies. Trap-vaccinate-release (TVR) is one alternate method that entails the use of live traps to capture targeted wildlife. Traps are distributed along grids in suitable, accessible habitats. Captured animals are identified by tagging, vaccinated parenterally with an inactivated commercial vaccine, sexed, aged, weighed, and released at the site of capture. Since 2001, cross-species transmission of rabies virus from bats to mesocarnivores, such as skunks, has been detected in Flagstaff, Arizona. During the past two decades, TVR has been used as a safe and effective method to prevent and control rabies during such events in this region.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Optimized and Simplified Technique for the Production and Culture of Precision-Cut Liver Slices.
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-22 DOI: 10.3791/67202
Dany Perocheau, Claire Duff, Sonam Gurung, Loukia Touramanidou, Garima Sharma, Julien Baruteau
{"title":"Optimized and Simplified Technique for the Production and Culture of Precision-Cut Liver Slices.","authors":"Dany Perocheau, Claire Duff, Sonam Gurung, Loukia Touramanidou, Garima Sharma, Julien Baruteau","doi":"10.3791/67202","DOIUrl":"https://doi.org/10.3791/67202","url":null,"abstract":"<p><p>This protocol presents a simple system for the creation and culture of Precision-cut Liver Slices (PCLS). PCLS contains all cells in an intact environment and, therefore, resembles a mini model of the whole organ. They enable the study of live tissues while replicating their complex phenotypes. This protocol allows the preparation of slices from mouse livers using a vibratome and standard laboratory equipment. Protocols for producing and culturing PCLS lack standardization and can vary quite drastically depending on the tissue of interest, the type of vibratome used, and the need for oxygen. These can be difficult to reproduce in some laboratories that have only access to a basic vibratome and common tissue culture facilities. We have put together a protocol focusing on the importance of some key steps within the varied protocols already available. This protocol, therefore, emphasizes the importance of the embedding method, the cutting orientation, a dynamic versus a static system, and the relevance of a minimum volume of culture. This protocol can be established and reproduced in a simple manner in most laboratories that have access to a basic tissue slicer. Taken together and following this protocol, PCLS can stay alive for a minimum of 4 days. PCLS is a simple, economical, and reproducible model to study pathophysiological and therapeutic screening for organs such as the liver.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142802544","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Innovative Approach to Minimize Serious Complications in Cosmetic Dermal Filling.
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-22 DOI: 10.3791/66540
Pengfei Li, Hui Lu
{"title":"Innovative Approach to Minimize Serious Complications in Cosmetic Dermal Filling.","authors":"Pengfei Li, Hui Lu","doi":"10.3791/66540","DOIUrl":"https://doi.org/10.3791/66540","url":null,"abstract":"<p><p>Cosmetic dermal filling represents a significant advance in aesthetic medicine, providing non-invasive treatments with immediate and durable outcomes. Despite the generally safe nature of dermal fillers and avoidable complications with proper technique and planning, they are still risky, with potential complications varying from mild and transient to severe and enduring. This study introduces innovative strategies designed to decrease the risk of serious complications, such as vascular occlusion. These include the use of ultrasound guidance, the application of ice, auxiliary gestures, effective withdrawal, etc. Throughout our research, we observed no severe complications like embolism, skin necrosis, or ocular issues. However, minor complications such as erythema, edema, and bruising were noted. It is important to recognize that while most severe complications can be avoided with meticulous technique and planning, the inherent risk of unexpected outcomes persists in any medical procedure. Our findings suggest that with these innovative strategies, the safety of cosmetic dermal filling can be significantly enhanced, thereby minimizing the risk of severe adverse effects.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142802608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Periorbital Placement of a Laser Doppler Probe for Cerebral Blood Flow Monitoring Prior to Middle Cerebral Artery Occlusion in Rodent Models.
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-22 DOI: 10.3791/66839
Denali C Dickson, Mitchell J Bartlet, Sharon Hom, Helena W Morrison
{"title":"Periorbital Placement of a Laser Doppler Probe for Cerebral Blood Flow Monitoring Prior to Middle Cerebral Artery Occlusion in Rodent Models.","authors":"Denali C Dickson, Mitchell J Bartlet, Sharon Hom, Helena W Morrison","doi":"10.3791/66839","DOIUrl":"https://doi.org/10.3791/66839","url":null,"abstract":"<p><p>Middle cerebral artery occlusion (MCAO) is the gold-standard method for preclinical modeling of ischemic stroke in rodents. However, successful occlusion is not guaranteed by even the most skilled surgical hands. Errors primarily occur when the filament is not placed at the correct depth and include instances of either no infarction or vessel perforation, which can cause death. Laser Doppler flowmetry (LDF) is a reliable technique that provides real-time feedback on regional cerebral blood flow (CBF) during the MCAO procedure. Here we demonstrate a rapid technique for periorbital placement of a laser Doppler probe for measurement of CBF in both mice and rats. Our rationale was to simplify LDF implementation, encouraging widespread usage for improved surgical reliability. The technique eliminates the need for skull thinning and specialized equipment, with placement at the periorbital region rather than dorsal placement, promoting efficiency and ease of adoption. The protocol described here encompasses presurgical preparations, periorbital Doppler probe placement, and post-operative care. Representative results include visual depictions of procedural elements along with representative LDF tracings illustrating successful MCAO surgeries, with instances of unsuccessful filament placement leading to complications. The protocol illustrates LDF in confirming proper filament placement and offers a simplified procedure compared to alternative methods.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142802547","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Screening Traditional Chinese Medicine Compounds for Inhibiting UCHL3 Activity Based on Molecular Docking and Deubiquitinating Enzyme Probe Technology.
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-22 DOI: 10.3791/67432
Shuxin Wang, Yaxin Qu, Yue Zhang, Zhu Fan, Bingnan Cui
{"title":"Screening Traditional Chinese Medicine Compounds for Inhibiting UCHL3 Activity Based on Molecular Docking and Deubiquitinating Enzyme Probe Technology.","authors":"Shuxin Wang, Yaxin Qu, Yue Zhang, Zhu Fan, Bingnan Cui","doi":"10.3791/67432","DOIUrl":"https://doi.org/10.3791/67432","url":null,"abstract":"<p><p>Deubiquitinating enzymes (DUBs) play a pivotal role in modulating ubiquitination homeostasis, with UCHL3 being an archetypal cysteine DUB intricately involved in a myriad of physiological and pathological processes. Therefore, developing small molecule inhibitors targeting Ubiquitin C-Terminal Hydrolase L3 (UCHL3) is of great significance. This protocol aims to establish a process for virtual screening and in vitro validation of small molecule inhibitors of cysteine DUB represented by UCHL3. Firstly, potential inhibitors of UCHL3 are virtually screened using molecular docking technology, and the interaction between drugs and protein targets is visualized. Subsequently, the effectiveness of the screened drug, Danshensu, is verified through in vitro activity inhibition assays. Ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) and hemagglutinin-ubiquitin-vinyl sulfone (HA-Ub-VS) are used as probes for in vitro activity testing, as they can competitively bind to DUB with small molecule inhibitors to assess the activity of UCHL3. The results indicate that Danshensu has a good binding affinity with UCHL3 in molecular docking, and it can competitively inhibit the activity of UCHL3 with HA-Ub-VS. These findings provide important references for further research and development of therapeutic drugs targeting UCHL3.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142802781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Observational Fear as a Model of Affective Empathy in Mice.
IF 1.2 4区 综合性期刊
Jove-Journal of Visualized Experiments Pub Date : 2024-11-22 DOI: 10.3791/67130
Hyoin Lee, Dahm So, Jiye Choi, Seungmoon Jung, Sehoon Keum
{"title":"Observational Fear as a Model of Affective Empathy in Mice.","authors":"Hyoin Lee, Dahm So, Jiye Choi, Seungmoon Jung, Sehoon Keum","doi":"10.3791/67130","DOIUrl":"https://doi.org/10.3791/67130","url":null,"abstract":"<p><p>Empathy, characterized by the ability to recognize and share the emotions of others, plays a fundamental role in shaping social interactions. It allows individuals to respond to the emotional states of others, promoting prosocial behaviors and social bonding. Observational fear is a fundamental aspect of affective empathy, where an observer witnesses a demonstrator undergoing aversive experiences and subsequently exhibits fear behaviors. This socially induced vicarious freezing response in observers, known as emotional contagion or affect sharing, is regarded as an indicator of empathy-like traits in rodents. Here, we present a protocol that delineates the assessment of vicarious freezing responses in observers exposed to conspecific demonstrators receiving aversive foot shocks. The utilization of observational fear assays in rodents has become a widely adopted method for studying the neural mechanisms underlying affective empathy. Given the universality of observational fear across mammals, this methodology further contributes to advancing our understanding of the neural substrates of empathy in humans.</p>","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142802514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"综合性期刊","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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