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Jilin University

13 ARTICLES PUBLISHED IN JoVE

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Biology

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)
Zach Hensel 1, Xiaona Fang 1,2,3, Jie Xiao 1
1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, 2Changchun Institute of Applied Chemistry, Chinese Academy of Sciences , 3Department of Physics, Jilin University

We describe a fluorescence microscopy method, Co-Translational Activation by Cleavage (CoTrAC), to image the production of protein molecules in live cells with single-molecule precision without perturbing the protein's functionality. This method has been used to follow the stochastic expression dynamics of a transcription factor, the λ repressor CI 1.

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Biology

Real-time Analyses of Retinol Transport by the Membrane Receptor of Plasma Retinol Binding Protein
Riki Kawaguchi 1, Ming Zhong 1, Hui Sun 1
1Department of Physiology, Jules Stein Eye Institute and Howard Hughes Medical Institute, University of California, Los Angeles

Here we describe an optimized technique to produce high-quality vitamin A/RBP complex and two real-time monitoring techniques to study vitamin A transport by STRA6, the RBP receptor.

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Developmental Biology

The C. elegans Intestine As a Model for Intercellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis at the Single-cell Level: Labeling by Antibody Staining, RNAi Loss-of-function Analysis and Imaging
Nan Zhang 1,2, Liakot A Khan 1, Edward Membreno 1, Gholamali Jafari 1, Siyang Yan 1, Hongjie Zhang 1,3, Verena Gobel 1
1Mucosal Immunology and Biology Research Center, Developmental Biology and Genetics Core, Massachusetts General Hospital, Harvard Medical School, 2College of Life Sciences, Jilin University, 3Faculty of Health Sciences, University of Macau

The transparent C. elegans intestine can serve as an "in vivo tissue chamber" for studying apicobasal membrane and lumen biogenesis at the single-cell and subcellular level during multicellular tubulogenesis. This protocol describes how to combine standard labeling, loss-of-function genetic/RNAi and microscopic approaches to dissect these processes on a molecular level.

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Developmental Biology

The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging
Nan Zhang 1,2, Edward Membreno 1, Susan Raj 1, Hongjie Zhang 1,3, Liakot A Khan 1, Verena Gobel 1
1Mucosal Immunology and Biology Research Center, Developmental Biology and Genetics Core, Massachusetts General Hospital for Children, Harvard Medical School, 2College of Life Sciences, Jilin University, 3Faculty of Health Sciences, University of Macau

The C. elegans excretory canal is a unique single-cell model for the visual in vivo analysis of de novo polarized membrane biogenesis. This protocol describes a combination of standard genetic/RNAi and imaging approaches, adaptable for the identification and characterization of molecules directing unicellular tubulogenesis, and apical membrane and lumen biogenesis.

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Developmental Biology

Evaluation of Injury-induced Senescence and In Vivo Reprogramming in the Skeletal Muscle
Coralie Cazin 1, Aurelie Chiche 1, Han Li 1
1Cellular Plasticity & Disease Modelling, Department of Developmental & Stem Cell Biology, CNRS UMR 3738, Institut Pasteur

Here we present a detailed protocol to detect both senescent and pluripotent stem cells in the skeletal muscle upon injury while inducing in vivo reprogramming. This method is suitable for evaluating the role of cellular senescence during tissue regeneration and reprogramming in vivo.

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Cancer Research

Selecting Multiple Biomarker Subsets with Similarly Effective Binary Classification Performances
Xin Feng 1, Shaofei Wang 1, Quewang Liu 1, Han Li 2, Jiamei Liu 2, Cheng Xu 2, Weifeng Yang 2, Yayun Shu 2, Weiwei Zheng 1, Bingxin Yu 3, Mingran Qi 4, Wenyang Zhou 1, Fengfeng Zhou 1
1College of Computer Science and Technology, and Key Laboratory of Symbolic Computation and Knowledge Engineering of Ministry of Education, Jilin University, 2College of Software, Jilin University, 3Ultrasonography Department, China-Japan Union Hospital of Jilin University, 4Department of Pathogenobiology, College of Basic Medical Science, Jilin University

Existing algorithms generate one solution for a biomarker detection dataset. This protocol demonstrates the existence of multiple similarly effective solutions and presents a user-friendly software to help biomedical researchers investigate their datasets for the proposed challenge. Computer scientists may also provide this feature in their biomarker detection algorithms.

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Medicine

Intra-Operative Neural Monitoring of Thyroid Surgery in a Porcine Model
Che-Wei Wu 1,2,3, Tzu-Yen Huang 2, Hui-Chun Chen 4, Hsiu-Ya Chen 5, Tsung-Yi Tsai 2, Pi-Ying Chang 5, Yi-Chu Lin 2, Hsin-Yi Tseng 2, Pao-Chu Hun 6, Xiaoli Liu 7, Hui Sun 7, Gregory W. Randolph 8, Gianlorenzo Dionigi 9, Feng-Yu Chiang 2,3, I-Cheng Lu 5,10,11
1Department of Otorhinolaryngology, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung Medical University, 2Department of Otorhinolaryngology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, 3Department of Otorhinolaryngology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, 4Department of Nursing, Kaohsiung Medical University Hospital, Kaohsiung Medical University, 5Department of Anesthesiology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, 6Laboratory Animal Center, Kaohsiung Medical University Hospital, Kaohsiung Medical University, 7Department of Thyroid and Parathyroid Surgery, China-Japan Union Hospital and Jilin Provincial Key Laboratory of Surgical Translational Medicine, Jilin University, 8Division of Thyroid and Parathyroid Endocrine Surgery, Department of Otolaryngology, Massachusetts Eye and Ear Infirmary; Division of Surgical Oncology, Department of Surgery, Massachusetts General Hospital; Department of Otology and Laryngology, Harvard Medical School, 9Division for Endocrine Surgery, Department of Human Pathology in Adulthood and Child-hood "G. Barresi", University Hospital G. Martino, University of Messina, 10Department of Anesthesiology, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung Medical University, 11Department of Anesthesiology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University

This study aims to develop a standard protocol of intra-operative neural monitoring of thyroid surgery in a porcine model. Here, we present a protocol to demonstrate general anesthesia, to compare different types of electrodes, and to investigate the electrophysiological characteristics of the normal and injured recurrent laryngeal nerves.

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Environment

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans
Shan Gao *1, Weiyang Chen *2, Nan Zhang 1, Chi Xu 3, Haiming Jing 1,4, Wenjing Zhang 1,4, Gaochao Han 1,4, Matthew Flavel 5, Markandeya Jois 5, Yingxin Zeng 1, Jing-Dong J. Han 3, Bo Xian 3, Guojun Li 1,4
1Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Prevention and Control/Beijing Center of Preventive Medicine Research, China, 2College of Computer Science and Technology, Qilu University of Technology(Shandong Academy of Sciences), China, 3Key Laboratory of Computational Biology, CAS Center for Excellence in Molecular Cell Science, Collaborative Innovation Center for Genetics and Developmental Biology, Chinese Academy of Sciences-Max Planck Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, China, 4Beijing Key Laboratory of Environmental Toxicology, School of Public Health, Capital Medical University, China, 5School of Life Sciences, La Trobe University, Australia

A quantitative method has been developed to identify and predict the acute toxicity of chemicals by automatically analyzing the phenotypic profiling of Caenorhabditis elegans. This protocol describes how to treat worms with chemicals in a 384-well plate, capture videos, and quantify toxicological related phenotypes.

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Medicine

Establishing In Situ Closed Circuit Perfusion of Lower Abdominal Organs and Hind Limbs in Mice
Ping Ren 1,2,3, Chunyan Yang 2,4, Laren A. Lofchy 2, Guankui Wang 2, Fangfang Chen *2,4,5, Dmitri Simberg *2
1Department of Thoracic Surgery, The First Hospital of Jilin University, 2The Skaggs School of Pharmacy and Pharmaceutical Sciences, Department of Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, 3Key Laboratory of Zoonoses Research, Ministry of Education, Jilin University, 4Department of Cardiology, China-Japan Union Hospital of Jilin University, 5Department of Gastrointestinal, Colorectal and Anal Surgery, China-Japan Union Hospital of Jilin University

A protocol is described for in situ perfusion of the mouse lower body, including the bladder, the prostate, sex organs, bone, muscle and foot skin.

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Neuroscience

Craniotomy Procedure for Visualizing Neuronal Activities in Hippocampus of Behaving Mice
YangDong Wang 1,2,3,4, DanYang Zhu 5, BaoYue Liu 1,2,3,6, Kiryl D. Piatkevich 1,2,3
1School of Life Sciences, Westlake University, 2Westlake Laboratory of Life Sciences and Biomedicine, 3Institute of Basic Medical Sciences, Westlake Institute for Advanced Study, 4School of Basic Medical Sciences, Xi'an JiaoTong University, 5Office of Public Affairs, Westlake University, 6School of Pharmaceutical Sciences, Jilin University

This article demonstrates the preparation of a custom-made imaging window supplemented with infusion cannula and its implantation onto the CA1 region of the hippocampus in mice.

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Genetics

Transient Transduction of the Strobilated Forms of Echinococcus granulosus
Mohammad Ali Mohammadi 1, Ali Afgar 2, Ashkan Faridi 3, Seyed Mohammad Mousavi 2, Ali Derakhshani 2, Mehdi Borhani 4, Majid Fasihi Harandi 2
1Student Research Committee, School of Medicine, Kerman University of Medical Sciences, 2Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, 3Department of Parasitology and Mycology, Faculty of Medicine, Kurdistan University of Medical Sciences, 4State Key Laboratory for Zoonotic Diseases, Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University

We describe a rapid transient transduction technique in different developmental stages of Echinococcus granulosus using third-generation lentiviral vectors.

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Neuroscience

Free-floating Immunostaining of Mouse Brains
Longlong Tu 1, Nan Zhang 1,2,3, Kristine M Conde 1, Jonathan C Bean 1, Chunmei Wang 1, Yong Xu 1,4
1USDA/ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 2Department of Endocrinology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 3Hubei Provincial Clinical Research Center for Diabetes and Metabolic Disorder, 4Department of Molecular and Cellular Biology, Baylor College of Medicine

This protocol describes an efficient and reproducible approach for mouse brain histological studies, including perfusion, brain sectioning, free-floating immunostaining, tissue mounting, and imaging.

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JoVE Core

Direct Reprogramming of Mouse Fibroblasts into Melanocytes
Yi-Xuan Zhang 1,2, Li-Ping Liu 1,2, Ming Jin 1,2, Hui Sun 1,2, Han-Lin Zhang 1,2, Yu-Mei Li 1,2
1Institute for Regenerative Medicine, Jiangsu University, 2Department of Dermatology, Affiliated Hospital of Jiangsu University

Here, we describe an optimized direct reprogramming system for melanocytes and a high-efficiency, concentrated virus packaging system that ensures smooth direct reprogramming.

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