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Agency for Science, Technology and Research

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Generation of Immature, Mature and Tolerogenic Dendritic Cells with Differing Metabolic Phenotypes

Wen Jing Sim¹, Frano Malinarich¹, Anna-Marie Fairhurst², John Edward Connolly^1,3

¹Translational Immunology, Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, ²Singapore Immunology Network, ³Institute of Biomedical Studies, Baylor University

Immature dendritic cells can be selectively differentiated into tolerogenic or mature dendritic cells to regulate the balance between immunity and tolerance. This work presents a means to generate from immature monocyte derived dendritic cells (moDCs), in vitro tolerogenic and mature moDCs that differ in metabolic phenotypes.

Behavior

Fully Automated Leg Tracking in Freely Moving Insects using Feature Learning Leg Segmentation and Tracking (FLLIT)

Animesh Banerjee¹, Shuang Wu², Li Cheng³, Sherry Shiying Aw¹

¹Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, ²Bioinformatics Institute, Agency for Science, Technology and Research, ³Department of Electrical and Computer Engineering, University of Alberta

We describe detailed protocols for using FLLIT, a fully automated machine learning method for leg claw movement tracking in freely moving Drosophila melanogaster and other insects. These protocols can be used to quantitatively measure subtle walking gait movements in wild type flies, mutant flies and fly models of neurodegeneration.

Neuroscience

Visualizing the Calcitonin Gene-Related Peptide Immunoreactive Innervation of the Rat Cranial Dura Mater with Immunofluorescence and Neural Tracing

Jia Wang^*1, Dongsheng Xu^*1, Jingjing Cui¹, Chen She¹, Hui Wang¹, Shuang Wu¹, Ling Zou¹, Jianliang Zhang¹, Wanzhu Bai¹

¹Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

Here we present a protocol to visualize spatial correlation of calcitonin gene-related peptide (CGRP)-immunoreactive nerve fibers and blood vessels in the cranial dura mater using immunofluorescence and fluorescent histochemistry with CGRP and phalloidin, respectively. In addition, the origin of these nerve fibers was retrograde traced with a fluorescent neural tracer.

Neuroscience

Visualizing the Morphological Characteristics of Neuromuscular Junction in Rat Medial Gastrocnemius Muscle

Jingjing Cui^*1, Shuang Wu^*1, Jia Wang¹, Yuqing Wang¹, Yuxin Su¹, Dongsheng Xu¹, Yihan Liu¹, Junhong Gao¹, Xianghong Jing¹, Wanzhu Bai¹

¹Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences

The protocol shows a method to examine spatial correlation among the pre-synaptic terminals, post-synaptic receptors, and peri-synaptic Schwann cells in the rat medial gastrocnemius muscle using fluorescent immunohistochemistry with different biomarkers, namely, neurofilament 200, vesicular acetylcholine transporter, alpha-bungarotoxin, and S100.