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Ben-Gurion University of the Negev

21 ARTICLES PUBLISHED IN JoVE

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Biology

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamAΔ41
Natalie Zeytuni 1, Raz Zarivach 1
1Department of Life Sciences and National Institute for Biotechnology in the Negev, Ben-Gurion University

MamA is a unique Magnetosome associated protein which was shown to be involved in magnetosome activation. Here we present the purification protocol of MamA deletion mutant (MamAΔ41) from M. magneticum AMB-1.

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Biology

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism
Ido Karady 1, Anna Frumkin 1, Shiran Dror 1, Netta Shemesh 1, Nadav Shai 1, Anat Ben-Zvi 1
1Department of Life Sciences, National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev

To study the relationship between protein homeostasis, stress and aging, we monitored changes in protein folding by following protein dysfunction, protein localization in the cell and protein stability at the organismal, cellular and protein levels, using the genetically tractable metazoan Caenorhabditis elegans as a model system.

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Neuroscience

Telomerase Activity in the Various Regions of Mouse Brain: Non-Radioactive Telomerase Repeat Amplification Protocol (TRAP) Assay
Yossi Grin 1, Tamar Admoni 1, Esther Priel 1
1The Shraga Segal Department of Immunology, Microbiology & Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev

Telomerase is expressed in the neonatal brain and also in distinct regions of the adult brain. We present a non-toxic time saving TRAP assay for the analysis of telomerase activity in various regions of the mouse brain and detection of differences in telomerase activity between male and female mouse brains.

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Engineering

High-speed Continuous-wave Stimulated Brillouin Scattering Spectrometer for Material Analysis
Itay Remer 1, Lear Cohen 1, Alberto Bilenca 1,2
1Biomedical Engineering Department, Ben-Gurion University of the Negev, 2Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev

We describe the construction of a rapid continuous-wave-stimulated-Brillouin-scattering (CW-SBS) spectrometer. The spectrometer employs single-frequency diode-lasers and an atomic vapor notch-filter to acquire transmission spectra of turbid/non-turbid samples with high spectral-resolution at speeds up to 100-fold faster than those of existing CW-SBS spectrometers. This improvement enables high-speed Brillouin material analysis.

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Neuroscience

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells
Tom Shani 1, Moshe Levy 1, Adrian Israelson 1,2
1Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, 2The Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev

This protocol describes a method to sensitively measure the nucleocytoplasmic transport rate within motor neuron-like NSC-34 cells by quantifying the real-time change in the nuclear import of a NLS-NES-GFP protein.

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Behavior

A New Method for Inducing a Depression-Like Behavior in Rats
Vladimir Zeldetz *1, Dmitry Natanel *2, Matthew Boyko 2, Alexander Zlotnik 2, Honore N. Shiyntum 3, Julia Grinshpun 2, Dmitry Frank 2, Ruslan Kuts 2, Evgeni Brotfain 2, Jochanan Peiser 2
1Department of Emergent Medicine, Soroka University Medical Center, Ben-Gurion University of the Negev, 2Division of Anesthesiology and Critical Care, Soroka Medical Center, Ben-Gurion University of the Negev, 3Department of Biophysics and Biochemistry, Oles' Honchar Dnipro National University

This protocol describes a new model by which healthy rats could contract depression over a given time periodthrough contagion by exposure to chronic unpredictable stressed (CUS) rats.

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Biochemistry

A Colorimetric Method for Measuring Iron Content in Plants
Jonas C. Gitz 1,2, Noy Sadot 1, Michele Zaccai 1, Raz Zarivach 1,2
1Department of Life Sciences, Ben-Gurion University of the Negev, 2National Institute for Biotechnology in the Negev (NIBN)

We present a simple and reliable protocol for measuring iron content in plant tissues using the colorimetric Prussian Blue method.

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Behavior

A Middle Cerebral Artery Occlusion Technique for Inducing Post-stroke Depression in Rats
Ruslan Kuts *1, Israel Melamed *2, Honore N. Shiyntum 3, Dmitry Frank 1, Julia Grinshpun 1, Alexander Zlotnik 1, Evgeni Brotfain 1, Michael Dubilet 1, Dmitry Natanel 1, Matthew Boyko 1
1Department of Anesthesiology and Critical Care, Soroka University Medical Center, Faculty of Health Sciences, Ben-Gurion University of the Negev, 2Department of Neurosurgery, Soroka University Medical Center, Faculty of Health Sciences, Ben-Gurion University of the Negev, 3Department of Biophysics and Biochemistry, Faculty of Biology, Ecology, and Medicine, Oles Honchar Dnipro National University

Here we present a protocol to induce post-stroke depression in rats by occluding the middle cerebral artery via the internal carotid artery. We use the Porsolt forced swim test and the sucrose preference test to confirm and evaluate induced depressive moods.

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Biochemistry

DNA Sequence Recognition by DNA Primase Using High-Throughput Primase Profiling
Stefan Ilic 1, Shira Cohen 1, Ariel Afek 2, Raluca Gordan 2, David B. Lukatsky 1, Barak Akabayov 1
1Department of Chemistry, Ben-Gurion University of the Negev, 2Center for Genomic and Computational Biology, Department of Biostatistics and Bioinformatics, Duke University

Protein binding microarray (PBM) experiments combined with biochemical assays link the binding and catalytic properties of DNA primase, an enzyme that synthesizes RNA primers on template DNA. This method, designated as high-throughput primase profiling (HTPP), can be used to reveal DNA-binding patterns of a variety of enzymes.

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Cancer Research

In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System
Manu Prasad *1,2, Sankar Jagadeeshan *1,2, Maurizio Scaltriti 3, Irit Allon 2,4, Moshe Elkabets 1,2
1The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, 2Faculty of Health Sciences, Ben-Gurion University of the Negev, 3Human Oncology & Pathogenesis Program (HOPP), Memorial Sloan Kettering Cancer Center, 4Institute of Pathology, Barzilai University Medical Center

Development of murine models with specific genes mutated in head and neck cancer patients is required for understanding of neoplasia. Here, we present a protocol for in vitro transformation of primary murine tongue cells using an adeno-associated virus-Cas9 system to generate murine HNC cell lines with specific genomic alterations.

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Neuroscience

Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish
Lear Cohen *1,2, Ehud Vinepinsky *2,3, Ronen Segev 1,2,3
1Department of Biomedical Engineering, Ben-Gurion University of the Negev, 2Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, 3Department of Life Sciences, Ben-Gurion University of the Negev

A novel wireless technique for recording extracellular neural signals from the brain of freely swimming goldfish is presented. The recording device is composed of two tetrodes, a microdrive, a neural data logger, and a waterproof case. All parts are custom-made except for the data logger and its connector.

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Medicine

Inducing Acute Liver Injury in Rats via Carbon Tetrachloride (CCl4) Exposure Through an Orogastric Tube
Dmitry Frank *1, Shiri Savir *1, Benjamin F. Gruenbaum 2, Israel Melamed 3, Julia Grinshpun 1, Ruslan Kuts 1, Boris Knyazer 4, Alexander Zlotnik 1, Max Vinokur 1, Matthew Boyko 1
1Department of Anesthesiology and Critical Care, Soroka Medical Center, Ben-Gurion University of the Negev, 2Department of Anesthesiology, Yale University School of Medicine, 3Department of Neurosurgery, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 4Department of Ophthalmology, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev

This protocol describes a common and feasible method of inducing acute liver injury (ALI) via CCl4 exposure through an orogastric tube. CCl4 exposure induces ALI through the formation of reactive oxygen species during its biotransformation in the liver. This method is used to analyze the pathophysiology of ALI and examine different hepatoprotective strategies.

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Developmental Biology

Generation of a Human iPSC-Based Blood-Brain Barrier Chip
Srikanth Jagadeesan 1,2,3, Michael J. Workman 4, Anna Herland 5,6, Clive N. Svendsen 4, Gad D. Vatine 1,2,3
1The Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, 2The Regenerative Medicine and Stem Cell (RMSC) Research Center, Ben-Gurion University of the Negev, 3The Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, 4The Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, 5Division of Micro and Nanosystems, KTH Royal Institute of Technology, 6AIMES, Department of Neuroscience, Karolinska Institutet

The blood-brain barrier (BBB) is a multicellular neurovascular unit tightly regulating brain homeostasis. By combining human iPSCs and organ-on-chip technologies, we have generated a personalized BBB chip, suitable for disease modeling and CNS drug penetrability predictions. A detailed protocol is described for the generation and operation of the BBB chip.

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Neuroscience

Laser-Induced Brain Injury in the Motor Cortex of Rats
Ruslan Kuts *1, Israel Melamed *2, Honore N. Shiyntum 3, Benjamin F. Gruenbaum 4, Dmitry Frank 1, Boris Knyazer 5, Dmitry Natanel 1, Olena Severynovska 3, Max Vinokur 1, Matthew Boyko 1
1Division of Anesthesiology and Critical Care, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 2Department of Neurosurgery, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 3Department of Biochemistry and Physiology, Faculty of Biology, Ecology, and Medicine, Oles Honchar Dnipro National University, 4Department of Anesthesiology, Yale University School of Medicine, 5Department of Ophthalmology, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev

The protocol presented here shows a technique to create a rodent model of brain injury. The method described here uses laser irradiation and targets motor cortex.

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Biology

Using Caenorhabditis elegans to Screen for Tissue-Specific Chaperone Interactions
Shiran Dror 1, Tomer D. Meidan 1, Ido Karady 1, Anat Ben-Zvi 1
1Department of Life Sciences, Ben-Gurion University of the Negev

To study chaperone-chaperone and chaperone-substrate interactions, we perform synthetic interaction screens in Caenorhabditis elegans using RNA interference in combination with mild mutations or over-expression of chaperones and monitor tissue-specific protein dysfunction at the organismal level.

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Neuroscience

Induction of Diffuse Axonal Brain Injury in Rats Based on Rotational Acceleration
Dmitry Frank *1, Israel Melamed *2, Benjamin F. Gruenbaum 3, Julia Grinshpun 1, Ruslan Kuts 1, Rachel Shvartsur 4, Abed N. Azab 4, Mohamad H. Assadi 5, Max Vinokur 1, Matthew Boyko 1
1Division of Anesthesia and Critical Care, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 2Department of Neurosurgery, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 3Department of Anesthesiology, Yale University School of Medicine, 4Recanati School for Community Health Professions, Faculty of Health Sciences, Ben-Gurion University of the Negev, 5Department of microbiology and immunology, Faculty of Health Sciences, Ben-Gurion University of the Negev

This protocol validates a reliable, easy-to-perform and reproducible rodent model of brain diffuse axonal injury (DAI) that induces widespread white matter damage without skull fractures or contusions.

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Neuroscience

Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples
Dmitry Frank *1, Benjamin F. Gruenbaum *2, Julia Grinshpun 1, Israel Melamed 3, Olena Severynovska 4, Ruslan Kuts 1, Michael Semyonov 1, Evgeni Brotfain 1, Alexander Zlotnik 1, Matthew Boyko 1
1Department of Anesthesiology and Critical Care, Soroka Medical Center, Ben-Gurion University of the Negev, 2Department of Anesthesiology, Yale University School of Medicine, 3Department of Neurosurgery, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 4Department of Physiology, Faculty of Biology, Ecology and Medicine, Dnepropetrovsk State University

This protocol describes a novel technique of measuring the three most important parameters of ischemic brain injury on the same set of rodent brain samples. Using only one brain sample is highly advantageous in terms of ethical and economic costs.

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Behavior

A Complex Diving-For-Food Task to Investigate Social Organization and Interactions in Rats
Benjamin F. Gruenbaum *1, Dmitry Frank *2, Shiri Savir 2, Honore N. Shiyntum 3, Ruslan Kuts 2, Max Vinokur 2, Israel Melamed 4, Michael Dubilet 2, Alexander Zlotnik 2, Matthew Boyko 2
1Department of Anesthesiology and Perioperative Medicine, Mayo Clinic, 2Division of Anesthesiology and Critical Care, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 3Department of Biophysics and Biochemistry, Oles Honchar Dnipro National University, 4Department of Neurosurgery, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev

This protocol describes a method of examining social hierarchy in a rat model. Rats perform a complex diving-for-food task in which they form a distinct hierarchy according to their willingness to dive underwater and swim to obtain a food pellet. This method is used to understand decision making and social relationships among highly social animals in small groups.

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Neuroscience

A Metric Test for Assessing Spatial Working Memory in Adult Rats Following Traumatic Brain Injury
Dmitry Frank *1, Benjamin F. Gruenbaum *2, Israel Melamed 3, Julia Grinshpun 1, Yair Benjamin 1, Ievgeni Vzhetson 1, Nadia Kravchenko 4, Michael Dubilet 1, Matthew Boyko *1, Alexander Zlotnik *1
1Department of Anesthesiology and Critical Care, Soroka Medical Center, Ben-Gurion University of the Negev, 2Department of Anesthesiology and Perioperative Medicine, Mayo Clinic, 3Department of Neurosurgery, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 4Department of Physiology, Faculty of Biology, Ecology and Medicine, Dnepropetrovsk State University

Traumatic brain injury (TBI) is commonly associated with memory impairment. Here, we present a protocol to assess spatial working memory after TBI via a metric task. A metric test is a useful tool to study spatial working memory impairment after TBI.

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Behavior

Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury
Dmitry Frank *1, Benjamin F. Gruenbaum *2, Michael Semyonov 1, Yair Binyamin 1, Olena Severynovska 3, Ron Gal 1, Amit Frenkel 1, Boris Knazer 4, Matthew Boyko 1, Alexander Zlotnik 1
1Division of Anesthesiology and Critical Care, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev, 2Department of Anesthesiology and Perioperative Medicine, Mayo Clinic, 3Department of Biochemistry and Physiology of the Faculty of Biology and Ecology, Oles Gonchar of the Dnipro National University, 4Department of Ophthalmology, Soroka University Medical Center and the Faculty of Health Sciences, Ben-Gurion University of the Negev

The present protocol describes a rat model of fluid percussion-induced traumatic brain injury followed by a series of behavioral tests to understand the development of dominant and submissive behavior. Using this model of traumatic brain injury in conjunction with specific behavioral tests enables the study of social impairments following brain injury.

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Biology

Imaging Flow Cytometry to Study Microbial Autoaggregation
Ronit Suissa 1, Uzi Hadad 2, Michael Meijler 1, Ilana Kolodkin-Gal 3,4
1Department of Chemistry, Ben-Gurion University of the Negev, 2Ilse Kats Institute for Nanoscale Science & Technology, Ben-Gurion University of the Negev, 3Department of Plant Pathology and Microbiology, Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, 4The Scojen institute for synthetic biology, Reichman university

This protocol describes a quantitative approach to measure microbial autoaggregation using imaging flow cytometry.

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