University of Macau

7 ARTICLES PUBLISHED IN JoVE

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 Khan¹, Edward Membreno¹, Gholamali Jafari¹, Siyang Yan¹, Hongjie Zhang^1,3, Verena Gobel¹

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.

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 Membreno¹, Susan Raj¹, Hongjie Zhang^1,3, Liakot A Khan¹, Verena Gobel¹

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.

Immunology and Infection

Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1^-/- Mice

Yongli Zhang^*1,2, Hangjie Zhang^*2, Wenqiang Ma^*3, Kefang Liu^1,2, Min Zhao⁴, Yingze Zhao², Xuancheng Lu⁵, Fuping Zhang⁶, Xiangdong Li³, George F. Gao^1,2,4,6, William J. Liu^1,2

¹School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, ²NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, ³State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, ⁴Research Network of Immunity and Health (RNIH), Beijing Institutes of Life Science, Chinese Academy of Sciences, ⁵Laboratory Animal Center, Chinese Center for Disease Control and Prevention, ⁶CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences

A protocol to evaluate antigen-specific T-cell responses in the immunoprivileged organs of the Ifnar1^-/- murine model for the Zika virus (ZIKV) infection is described. This method is pivotal for investigating the cellular mechanisms of the protection and immunopathogenesis of ZIKV vaccines and is also valuable for their efficacy evaluation.

Behavior

Impact of High-intensity Interval Exercise and Moderate-Intensity Continuous Exercise on the Cardiac Troponin T Level at an Early Stage of Training

Haifeng Zhang^1,2, Jinlei Nie³, Zhaowei Kong⁴, Xiangui Zhu¹, Yang Liu¹, Qingde Shi³

¹Physical Education College, Hebei Normal University, ²Hebei Provincial Key Lab of Measurement and Evaluation in Human Movement and Bio-Information, ³School of Health Sciences and Sports, Macao Polytechnic Institute, ⁴Faculty of Education, University of Macau

Here, we present protocols of high-intensity interval and moderate-intensity continuous exercise to observe the response of circulating cardiac troponin T (cTnT) concentration to acute exercise over 10 days. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise.

Neuroscience

Conducting Concurrent Electroencephalography and Functional Near-Infrared Spectroscopy Recordings with a Flanker Task

Shi Yang Xu^1,2, Lai Ian Cheong², Yin Zhuang², Tania Alexandra Pinho Couto^1,2, Zhen Yuan^1,2

¹Centre for Cognitive and Brain Sciences, University of Macau, ²Faculty of Health Sciences, University of Macau

The present protocol describes how to perform concurrent EEG and fNIRS recordings and how to inspect the relationship between the EEG and fNIRS data.

Immunology and Infection

Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β₂-Microglobulin

Di Zhang^*1,2, Kefang Liu^*3,4, Dan Lu^*2,5, Pengyan Wang^1,2, Qingxu Zhang^1,2, Peipei Liu², Yingze Zhao², Yan Chai⁴, Jianxin Lyu¹, Jianxun Qi⁴, William J. Liu^1,2

¹School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, ²NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, ³Faculty of Health Sciences, University of Macau, ⁴CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, ⁵Savaid Medical School, University of Chinese Academy of Sciences

The protocol describes experimental methods to obtain stable major histocompatibility complex (MHC) class I through potential β₂-microglobulin (β₂m) substitutions from different species. The structural comparison of MHC I stabilized by homologous and heterologous β₂m were investigated.

Biology

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka (Oryzias latipes)

Muhammad Rahmad Royan¹, Shinji Kanda², Daichi Kayo³, Weiyi Song⁴, Wei Ge⁴, Finn-Arne Weltzien¹, Romain Fontaine¹

¹Physiology Unit, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, ²Laboratory of Physiology, Atmosphere and Ocean Research Institute, The University of Tokyo, ³Department of Biological Sciences, Graduate School of Science, The University of Tokyo, ⁴Centre of Reproduction, Development and Aging (CRDA), Faculty of Health Sciences, University of Macau

The article describes a quick protocol to gonadectomize and sample blood from the small teleost fish, using Japanese medaka (Oryzias latipes) as a model, to investigate the role of sex steroids in animal physiology.