Jove-Journal of Visualized Experiments最新文献_第4页

Establishing a Severe Corneal Inflammation Model in Rats Based on Corneal Epithelium Curettage Combined with Corneal Sutures.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-22 DOI: 10.3791/67305

Xiaoyu Tian, Meng Zhang, Yiming Wu, Liying Zhang, Lingli Zhang, Xueer Zheng, Shangkun Ou, Hao Gu

{"title":"Establishing a Severe Corneal Inflammation Model in Rats Based on Corneal Epithelium Curettage Combined with Corneal Sutures.","authors":"Xiaoyu Tian, Meng Zhang, Yiming Wu, Liying Zhang, Lingli Zhang, Xueer Zheng, Shangkun Ou, Hao Gu","doi":"10.3791/67305","DOIUrl":"https://doi.org/10.3791/67305","url":null,"abstract":"Corneal inflammation, especially severe corneal inflammation, plays a significant role in the development of corneal limbal stem cell dysfunction. Constructing appropriate animal models can help us focus on the effects of severe inflammation on corneal limbal stem cells. A 2 mm rust remover was used to remove the central corneal epithelium of Sprague Dawley (SD) rats to create an injury. Then, the central stroma of the cornea was sutured with nylon sutures to induce persistent inflammation. In this way, a corneal inflammation model with central corneal epithelium abrasion and central stroma suturing was constructed, which induced severe corneal inflammation. The changes in corneal inflammation and the condition of the limbal stem cells at 1, 3, and 7 days post-modeling were observed. On the 3rd day after modeling, the rats' corneal limbus was severely edematous, with obvious neovascularization and local hyperplasia, which are typical signs of limbal stem cell deficiency. By the 7th day, the corneal edema gradually worsened, and the neovascularization continued to increase. Through quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence staining, we found that the corneal epithelial inflammatory factors were significantly upregulated, the corneal epithelial differentiation was abnormal, the corneal epithelial stem cells were significantly reduced, and the cell proliferation and stemness had also decreased. Therefore, this model demonstrates that severe inflammation can induce limbal stem cell damage without directly damaging the limbal stem cells. The model is beneficial for observing the effects of severe inflammation on the biological mechanisms of stem cells and provides an ideal platform for studying the mechanisms of corneal epithelial stem cell dysfunction induced by inflammation.","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":"142802590","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

Erratum: Measuring Psoriasis Severity at Home. 勘误：在家中测量牛皮癣的严重程度。

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-21 DOI: 10.3791/6612

Courtney Carroll, Jonas A Adalsteinsson, Megan Prouty, Kristina Callis Duffin, Gerald G Krueger, Jessica A Walsh, Bing-Jian Feng

引用次数: 0

An Integrated Method for Crafting Flexible and Convenient Electrophysiological Optrodes for Multi-Region In Vivo Recording.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-21 DOI: 10.3791/67071

Yan Tao, Yuxin Zhao, Wenqi Zhong, Ruiqi Wu

{"title":"An Integrated Method for Crafting Flexible and Convenient Electrophysiological Optrodes for Multi-Region In Vivo Recording.","authors":"Yan Tao, Yuxin Zhao, Wenqi Zhong, Ruiqi Wu","doi":"10.3791/67071","DOIUrl":"https://doi.org/10.3791/67071","url":null,"abstract":"Epilepsy is a neurological disorder characterized by synchronized abnormal discharges involving multiple brain regions. Focal lesions facilitate the propagation of epileptic signals through associated neural circuits. Therefore, in vivo recording of local field potential (LFP) from the critical brain regions is essential for deciphering the circuits involved in seizure propagation. However, current methods for electrode fabrication and implantation lack flexibility. Here, we present a handy device designed for electrophysiological recordings (LFPs and electroencephalography [EEG]) across multiple regions. Additionally, we seamlessly integrated optogenetic manipulation and calcium signaling recording with LFP recording. Robust after-discharges were observed in several separate regions during epileptic seizures, accompanied by increasing calcium signaling. The approach used in this study offers a convenient and flexible strategy for synchronous neural recordings across diverse regions of the brain. It holds the potential for advancing research on neurological disorders by providing insights into the neural profiles of multiple regions involved in these disorders.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142802915","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 Real-Time Wearable Electromyography Measurement System for Small Animals.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/67418

Youngwoo Yoo, Ae-Jin Song, Sungho Lee, Myunghwan Choi, Ho-Sueb Song, Young-Joon Kim

{"title":"A Real-Time Wearable Electromyography Measurement System for Small Animals.","authors":"Youngwoo Yoo, Ae-Jin Song, Sungho Lee, Myunghwan Choi, Ho-Sueb Song, Young-Joon Kim","doi":"10.3791/67418","DOIUrl":"https://doi.org/10.3791/67418","url":null,"abstract":"The intramuscular electromyography (EMG) measurement method for experimental animals has been implemented in various ways. Among these methods, tethering cables to external measurement devices can restrict the movement of experimental animals, while implantable devices may cause unwanted side effects due to the constant presence of a device with considerable size and weight. To address these issues, we propose a low-cost, wireless, detachable EMG measurement system and experimental procedure. This article focuses on the surgical installation of intramuscular wire electrodes with small connectors and the development of the wireless system. Notably, in this system, only the wire electrodes are inserted into the animal's body. Using this system, EMG measurements can be easily performed by attaching the circuit system to a connector installed on the animal's back, with real-time monitoring achievable on a laptop. The proposed method is explained in a detailed, step-by-step manner, followed by a demonstration involving the insertion of intramuscular electrodes into the hindlimbs of a rat. A treadmill experiment is conducted for a locomotion study, and the resulting electrophysiological signals are subsequently obtained and analyzed.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774031","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

An Immunocompetent Murine Model for Laser Interstitial Thermal Therapy of Glioblastoma.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/67381

Jeremy Spence, Santhosh S Anandhan, Nimrat Kaur, Thatchawan Thanasupawat, Sabine Hombach-Klonisch, Thomas Klonisch

{"title":"An Immunocompetent Murine Model for Laser Interstitial Thermal Therapy of Glioblastoma.","authors":"Jeremy Spence, Santhosh S Anandhan, Nimrat Kaur, Thatchawan Thanasupawat, Sabine Hombach-Klonisch, Thomas Klonisch","doi":"10.3791/67381","DOIUrl":"https://doi.org/10.3791/67381","url":null,"abstract":"Glioblastoma (GB), the most aggressive form of primary brain cancer, accounts for approximately half of all high-grade primary brain tumors in adults and has no cure. Laser interstitial thermal therapy (LITT) is a Food and Drug Administration (FDA)-approved treatment for GB and is used in patients who may not be candidates for conventional surgical resection. While the clinical efficacy of LITT has been established, research beyond clinical case studies and case series is limited and hindered by the lack of an established animal model. This protocol uses C57BL/6 mice and syngeneic CT2A glioma cancer cell line to closely recapitulate human GB while also using a 1064 nm Neodymium-doped Yttrium Aluminum Garnet (Nd:YAG) laser, such as is used in one of the two FDA-approved LITT systems, providing excellent pre-clinical relevance. The successful establishment of this LITT murine model will provide a valuable platform for investigating the unique features of LITT ablation and its effects on the tumor microenvironment, potentially leading to improved therapeutic strategies.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774032","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

Fibro-Adipogenic Progenitor Isolation, Expansion, and Differentiation from the Spiny Mouse Model.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/66717

Bruce Lin, Hesham Soliman, Fabio M V Rossi, Marine Theret

{"title":"Fibro-Adipogenic Progenitor Isolation, Expansion, and Differentiation from the Spiny Mouse Model.","authors":"Bruce Lin, Hesham Soliman, Fabio M V Rossi, Marine Theret","doi":"10.3791/66717","DOIUrl":"https://doi.org/10.3791/66717","url":null,"abstract":"Due to its exceptional repair program, the spiny mouse is an emerging research model for regenerative medicine. Fibro-adipogenic progenitors are tissue-resident cells that are able to differentiate into adipocytes, fibroblasts, and chondrocytes. Fibro-adipogenic progenitors are fundamental for orchestrating tissue regeneration as they are responsible for extracellular matrix remodeling after injury. This study focuses on investigating the specific role of fibro-adipogenic progenitors in spiny mouse cardiac repair and skeletal muscle regeneration. To this end, a protocol has been optimized for the purification of spiny mouse fibro-adipogenic progenitors by flow cytometry from enzymatically dissociated skeletal and cardiac muscle. The population obtained from this protocol is capable of expanding in vitro, and can be differentiated to myofibroblasts and adipocytes. This protocol offers a valuable tool for researchers to examine the distinctive properties of spiny mouse, and to compare them to the Mus musculus. This will provide insights that could advance the understanding of regenerative mechanisms in this intriguing model.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142773791","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

Novel Quantification Protocol for Cardiovascular Calcification Progression Using Longitudinal MicroPET/MicroCT Images.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/66805

Isabel L Day, Mikayla Tamboline, Andrea Litwak, Andrea Sarabia, Yin Tintut, Linda L Demer, Shili Xu

{"title":"Novel Quantification Protocol for Cardiovascular Calcification Progression Using Longitudinal MicroPET/MicroCT Images.","authors":"Isabel L Day, Mikayla Tamboline, Andrea Litwak, Andrea Sarabia, Yin Tintut, Linda L Demer, Shili Xu","doi":"10.3791/66805","DOIUrl":"https://doi.org/10.3791/66805","url":null,"abstract":"Micro positron emission tomography (PET) and micro computed tomography (CT) imaging are powerful, ideal research tools for following the progression of cardiovascular calcification. Due to their non-invasive nature, small research animals can be imaged at multiple time points. The challenge lies in the accurate quantification of cardiovascular calcification. Here, we provide a protocol, using images from the later disease stages as a template, to accurately quantify the progression of cardiovascular calcification in longitudinal studies. The protocol involves 1) the alignment of the chest area in multiple images from the same animal during a longitudinal study as the first step, 2) the definition of a region of interest (ROI) situated within the heart and the aorta at the site of larger calcium deposits that become apparent in later images, and 3) simultaneous segmentation and quantification of calcium deposits across all images acquired during the longitudinal study. This streamlined method enhances the accuracy of image analysis in following the progression of cardiovascular calcification by improving the precision of ROI definition and reducing the variability associated with earlier techniques that analyze individual scans independently.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142773856","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

Retroviral CRISPR/Cas9-Mediated Gene Targeting for the Study of Th17 Differentiation in Vitro.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/66966

Zejin Cui, Pengkun Yuan, Zhishan Zhao, Fan Zhao, Linrong Lu

{"title":"Retroviral CRISPR/Cas9-Mediated Gene Targeting for the Study of Th17 Differentiation in Vitro.","authors":"Zejin Cui, Pengkun Yuan, Zhishan Zhao, Fan Zhao, Linrong Lu","doi":"10.3791/66966","DOIUrl":"https://doi.org/10.3791/66966","url":null,"abstract":"T helper cells that produce IL-17A, known as Th17 cells, play a critical role in immune defense and are implicated in autoimmune disorders. CD4 T cells can be stimulated with antigens and well-defined cytokine cocktails in vitro to mimic Th17 cell differentiation in vivo. Research has been conducted extensively on the Th17 differentiation regulation mechanisms using the in vitro Th17 polarization assay. Conventional Th17 polarization methods typically involve obtaining naïve CD4 T cells from genetically modified mice to study the effects of specific genes on Th17 differentiation and function. These methods can be time-consuming and costly and may be influenced by cell-extrinsic factors from the knockout animals. Thus, a protocol using retroviral transduction of guide RNA to introduce gene knockout in CRISPR/Cas9 knockin primary mouse T cells serves as a very useful alternative approach. This paper presents a protocol to differentiate naïve primary T cells into Th17 cells following retroviral-mediated gene targeting, as well as the subsequent flow cytometry analysis methods for assaying infection and differentiation efficiency.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142773862","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 Modified Inflammatory Pain Model to Study the Analgesic Effect in Mice.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/66701

Li Huiwen, Zheng Baiyang, Wei Yongkang, Jia Xue, Zhang Jiren, Zhang Pinyuan, Wang Tian

{"title":"A Modified Inflammatory Pain Model to Study the Analgesic Effect in Mice.","authors":"Li Huiwen, Zheng Baiyang, Wei Yongkang, Jia Xue, Zhang Jiren, Zhang Pinyuan, Wang Tian","doi":"10.3791/66701","DOIUrl":"https://doi.org/10.3791/66701","url":null,"abstract":"The hot plate test is widely used to evaluate analgesic effects on inflammatory pain in mice. A commonly used model of inflammatory pain was induced with an intraplantar injection of carrageenan in one hind paw. However, the findings from our laboratory showed that mice with a single-hind-paw injection of carrageenan lifted their paws to avoid thermal nociception during the hot plate test. Because of this response, previous injection method cannot accurately reflect the thermal pain threshold. Thus, we investigated a new method to avoid this issue. In the present study, we modified the previous method by injecting carrageenan into both hind paws to establish the model of inflammatory pain. The results demonstrated that both-hind-paw injection with carrageenan was sensitive and a better method to induce inflammatory pain when using the hot plate test than single-hind-paw injection. On the basis of these findings, we designed further experiments in which mice with either both-hind-paw or single-hind-paw injection of carrageenan were treated intragastrically with celecoxib (30 mg/kg). The results of the hot plate test showed that celecoxib augmented the thermal pain threshold in mice with both-hind-paw injection of carrageenan but not in mice with single-hind-paw injection of carrageenan. In summary, we developed a superior method to induce a model of inflammatory pain to evaluate analgesic effect.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774028","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

Ultrasound Assessment of Aortic and Carotid Artery Intima-media Thickness in Children and Adolescents.

IF 1.2 4区综合性期刊

Jove-Journal of Visualized Experiments Pub Date : 2024-11-15 DOI: 10.3791/67095

Reeja Nasir, Michael Skilton, Adrienne Gordon

{"title":"Ultrasound Assessment of Aortic and Carotid Artery Intima-media Thickness in Children and Adolescents.","authors":"Reeja Nasir, Michael Skilton, Adrienne Gordon","doi":"10.3791/67095","DOIUrl":"https://doi.org/10.3791/67095","url":null,"abstract":"Carotid intima-media thickness (IMT), measured using high-resolution B-mode ultrasonography, is a widely utilized surrogate marker of subclinical atherosclerosis, the pathophysiological process underlying most clinical cardiovascular disease events. Atherosclerosis is a gradual disease that originates early in life, thus, there has been increased interest in measuring carotid IMT in childhood and adolescence to assess structural change in the arterial vasculature in response to adverse exposures. However, the timing of atherosclerosis varies across the vascular tree. Primordial atherosclerotic lesions are present in the abdominal aorta as early as infancy, compared to mid-adolescence for the common carotid. Measurement of IMT at either site is susceptible to several technical challenges that need to be considered, especially in younger children. In this paper, we provide a detailed stepwise method for high-quality assessment of IMT of the abdominal aorta and common carotid artery in the young. We also provide insight into the appropriateness of either site when exploring the associations between early-life exposures and later-life cardiovascular disease.","PeriodicalId":48787,"journal":{"name":"Jove-Journal of Visualized Experiments","volume":" 213","pages":""},"PeriodicalIF":1.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142773866","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