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4 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Generation of Immature, Mature and Tolerogenic Dendritic Cells with Differing Metabolic Phenotypes
Wen Jing Sim 1, Frano Malinarich 1, Anna-Marie Fairhurst 2, 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.

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Behavior

Fully Automated Leg Tracking in Freely Moving Insects using Feature Learning Leg Segmentation and Tracking (FLLIT)
Animesh Banerjee 1, Shuang Wu 2, Li Cheng 3, Sherry Shiying Aw 1
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.

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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 Cui 1, Chen She 1, Hui Wang 1, Shuang Wu 1, Ling Zou 1, Jianliang Zhang 1, Wanzhu Bai 1
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.

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Neuroscience

Visualizing the Morphological Characteristics of Neuromuscular Junction in Rat Medial Gastrocnemius Muscle
Jingjing Cui *1, Shuang Wu *1, Jia Wang 1, Yuqing Wang 1, Yuxin Su 1, Dongsheng Xu 1, Yihan Liu 1, Junhong Gao 1, Xianghong Jing 1, Wanzhu Bai 1
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.

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