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17 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection
Jana Boerner 1, Tanja A. Godenschwege 1
1Department of Biological Sciences, Florida Atlantic University

An in vivo dissection of the adult Drosophila ventral nerve cord (VNC) is demonstrated. This particular dissection method causes little damage to the VNC allowing the subsequent labeling of the giant fiber neurons with fluorescent dye for high resolution imaging.

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Neuroscience

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System
Monica Mejia 1, Mari D. Heghinian 2, Alexandra Busch 1, Frank Marí 2, Tanja A. Godenschwege 1
1Department of Biological Sciences, Florida Atlantic University, 2Department of Chemistry & Biochemistry, Florida Atlantic University

A rapid in vivo assay to test for neuromodulatory compounds using the Giant Fiber System (GFS) of Drosophila melanogaster is described. Nanoinjections in the head of the animal along with electrophysiological recordings of the GFS can reveal bioactivity of compounds on neurons or muscles.

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Immunology and Infection

A Protocol to Infect Caenorhabditis elegans with Salmonella typhimurium
Jiuli Zhang 1, Kailiang Jia 1
1Department of Biological Sciences, Florida Atlantic University

C. elegans has emerged as a new genetic model to study host-pathogen interactions. Here we describe a protocol to infect C. elegans with Salmonella typhimurium coupled with the double-strand RNAi interference technique to examine the role of host genes in defense against Salmonella infection.

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Neuroscience

Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome
Ibrahim M. Shokry 1, John J. Callanan 1, John Sousa 2, Rui Tao 2
1Ross University School of Veterinary Medicine, 2Charles E. Schmidt College of Medicine, Florida Atlantic University

This protocol describes a rapid and simplified in situ hybridization method ideal forparaformaldehyde-prefixed brain, thus reducing the need for prolonged complex steps while using fresh frozen tissues. The method is validated using the identification of the serotonin 5-HT2A receptor gene htr2a in rats.

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Biology

Genome Editing in Astyanax mexicanus Using Transcription Activator-like Effector Nucleases (TALENs)
Johanna E. Kowalko 1, Li Ma 2, William R. Jeffery 3
1Genetics, Development and Cell Biology, Iowa State University, 2Department of Biological Sciences, University of Cincinnati, 3Department of Biology, University of Maryland

Gene-targeting mutagenesis is now possible in a wide range of organisms using genome editing techniques. Here, we demonstrate a protocol for targeted gene mutagenesis using transcription activator like effector nucleases (TALENs) in Astyanax mexicanus, a species of fish that includes surface fish and cavefish.

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Engineering

Measurement of Quantum Interference in a Silicon Ring Resonator Photon Source
Jeffrey A. Steidle 1, Michael L. Fanto 1,2, Stefan F. Preble 1, Christopher C. Tison 2,3,4, Gregory A. Howland 2, Zihao Wang 1, Paul M. Alsing 2
1Microsystems Engineering, Rochester Institute of Technology, 2Air Force Research Laboratory, Rome, NY, 3Department of Physics, Florida Atlantic University, 4Quanterion Solutions Incorporated

Silicon photonic chips have the potential to realize complex integrated quantum systems. Presented here is a method for preparing and testing a silicon photonic chip for quantum measurements.

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Medicine

Application of Granger Causality Analysis of the Directed Functional Connection in Alzheimer's Disease and Mild Cognitive Impairment
Mei Wang 1, Zhengluan Liao 2, Dewang Mao 3, Qi Zhang 1, Yumei Li 3, Enyan Yu 2, Zhongxiang Ding 3
1Zhejiang Chinese Medical University, 2Department of Psychiatry, Zhejiang Provincial People's Hospital, 3Department of Radiology, Zhejiang Provincial People's Hospital

Based on resting-state functional magnetic resonance imaging with Granger causality analysis, we investigated the alterations in the directed functional connectivity between the posterior cingulate cortex and whole brain in patients with Alzheimer's Disease (AD), patients with Mild Cognitive Impairment (MCI), and healthy controls.

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Behavior

An Automated T-maze Based Apparatus and Protocol for Analyzing Delay- and Effort-based Decision Making in Free Moving Rodents
Qi Zhang 1,2, Yuki Kobayashi 1, Hiromichi Goto 1, Shigeyoshi Itohara 1
1Laboratory of Behavioral Genetics, Center for Brain Science, RIKEN, 2Faculty of Human Science, University of Tsukuba

This article introduces an automated T-maze apparatus that we invented, and a protocol based on this apparatus for analyzing delay-based decision making and effort-based decision making in free moving rodents.

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Behavior

Behavioral Approaches to Studying Innate Stress in Zebrafish
Jacqueline S.R. Chin 1, Lydia T. Albert 2, Cody L. Loomis 1,3, Alex C. Keene 1,3, Erik R. Duboué 1,2,3
1Jupiter Life Science Initiative, Florida Atlantic University, 2Wilkes Honors College and Florida Atlantic University, 3Charles E. Schmidt College of Science, Florida Atlantic University

This manuscript describes a simple method to measure stress behaviorally in adult zebrafish. The approach takes advantage of the innate tendency that zebrafish prefer the bottom half of a tank when in a stressful state. We also describe methods for coupling the assay with pharmacology.

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JoVE Journal

Manipulation of Gene Function in Mexican Cavefish
Bethany A. Stahl 1,2, James B. Jaggard 1,2, Jacqueline S.R. Chin 1,2, Johanna E. Kowalko 2,3, Alex C. Keene 1,2, Erik R. Duboué 2,3
1Department of Biological Sciences, Florida Atlantic University, 2Jupiter Life Science Initiative, Florida Atlantic University, 3Harriet L. Wilkes Honors College, Florida Atlantic University

We describe approaches for the manipulation of genes in the evolutionary model system Astyanax mexicanus. Three different techniques are described: Tol2-mediated transgenesis, targeted manipulation of the genome using CRISPR/Cas9, and knockdown of expression using morpholinos. These tools should facilitate the direct investigation of genes underlying the variation between surface- and cave-dwelling forms.

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Behavior

Automated Measurements of Sleep and Locomotor Activity in Mexican Cavefish
James B. Jaggard 1,2, Evan Lloyd 1,2, Arthur Lopatto 2,3, Erik R. Duboue 2,3, Alex C. Keene 1,2
1Department of Biological Sciences, Florida Atlantic University, 2Jupiter Life Science Initiative, Florida Atlantic University, 3Harriet L. Wilkes Honors College, Florida Atlantic University

This protocol details the methodology for quantifying locomotor behavior and sleep in the Mexican cavefish. Previous analyses are extended to measure these behaviors in socially-housed fish. This system can be widely applied to study sleep and activity in other fish species.

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Behavior

Swimming Induced Paralysis to Assess Dopamine Signaling in Caenorhabditis elegans
Sirisha Kudumala 1, Serena Sossi 2, Lucia Carvelli 1,3,4
1Harriet Wilkes Honors College, Florida Atlantic University, John D MacArthur Campus, 2Integrative Biology and Neuroscience program, College of Science, Florida Atlantic University, 3Brain Institute, Florida Atlantic University, 4Department of Biomedical Science, Charles E. Schmidt College of Medicine, Florida Atlantic University

Swimming induced paralysis (SWIP) is a well-established behavioral assay used to study the underlying mechanisms of dopamine signaling in Caenorhabditis elegans (C. elegans). However, a detailed method to perform the assay is lacking. Here, we describe a step-by-step protocol for SWIP.

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Neuroscience

Measuring Membrane Lipid Turnover with the pH-sensitive Fluorescent Lipid Analog ND6
Shahriar Alamgir 1,2, Oliver B. Pelletier 1,2, Deborah Thomas 3, Vicente Rubio 3, Maciej J. Stawikowski 3, Qi Zhang 1,2
1The Brain Institute at Florida Atlantic University, 2Department of Biomedical Sciences, Florida Atlantic University, 3Department of Chemistry and Biochemistry, Florida Atlantic University

This protocol presents a fluorescence imaging method that uses a class of pH-sensitive lipid fluorophores to monitor lipid membrane trafficking during cell exocytosis and the endocytosis cycle.

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Biology

Inherent Dynamics Visualizer, an Interactive Application for Evaluating and Visualizing Outputs from a Gene Regulatory Network Inference Pipeline
Robert C. Moseley 1, Sophia Campione 1, Bree Cummins 2, Francis Motta 3, Steven B. Haase 1
1Department of Biology, Duke University, 2Department of Mathematical Sciences, Montana State University, 3Department of Mathematical Sciences, Florida Atlantic University

The Inherent Dynamics Visualizer is an interactive visualization package that connects to a gene regulatory network inference tool for enhanced, streamlined generation of functional network models. The visualizer can be used to make more informed decisions for parameterizing the inference tool, thus increasing confidence in the resulting models.

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Neuroscience

New Framework for Understanding Cross-Brain Coherence in Functional Near-Infrared Spectroscopy (fNIRS) Hyperscanning Studies
Hila Zahava Gvirts Provolovski 1, Mini Sharma 1, Itai Gutman 1, Anat Dahan 2, Yafeng Pan 3, Jacqueline Stotler 4, Teresa Wilcox 4
1The Department of Behavioral Sciences and Psychology, Ariel University, 2Braude College of Engineering, 3Department of Psychology and Behavioral Sciences, Zhejiang University, 4Department of Psychology, Florida Atlantic University

Wavelet transform coherence (WTC) is a common methodology for assessing the coupling between signals that is used in functional near-infrared spectroscopy (fNIRS) hyperscanning studies. A toolbox for assessing the directionality of the signal interaction is presented in this work.

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JoVE Journal

Amplitude-Modulated Electrodeformation to Evaluate Mechanical Fatigue of Biological Cells
Darryl Dieujuste 1, Adeleh Kazemi Alamouti 1, Hongyuan Xu 1, E Du 1,2
1Department of Ocean and Mechanical Engineering, Florida Atlantic University, 2Center for SMART Health, Florida Atlantic University

Presented here is a protocol for mechanical fatigue testing in the case of human red blood cells using an amplitude-modulated electrodeformation approach. This general approach can be used to measure the systematic changes in morphological and biomechanical characteristics of biological cells in a suspension from cyclic deformation.

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Neuroscience

Laser Cell Ablation in Intact Drosophila Larvae Reveals Synaptic Competition
Jana Boerner 1, Kelli Robbins 2, Rod Murphey 2
1Stiles-Nicholson Brain Institute, Florida Atlantic University, 2Department of Biological Sciences, Florida Atlantic University

This protocol demonstrates the laser cell ablation of individual neurons in intact Drosophila larvae. The method enables the study of the effect of reducing competition between neurons in the developing nervous system.

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