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University of Fribourg

22 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Behavioral Assessment of Manual Dexterity in Non-Human Primates
Eric Schmidlin *1, Mélanie Kaeser *1, Anne- Dominique Gindrat 1, Julie Savidan 1, Pauline Chatagny 1, Simon Badoud 1, Adjia Hamadjida 1, Marie-Laure Beaud 1, Thierry Wannier 1, Abderraouf Belhaj-Saif 1, Eric M. Rouiller 1
1Department of Medicine, University of Fribourg

As manual dexterity is a prerogative mainly of primates, behavioral tasks have been developed in macaque monkeys. Four reach and grasp prehension tasks, measuring hand manipulation ability and force, allow to establish functional recovery after a lesion of the central nervous system and to test the effect of a treatment.

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Medicine

Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury
Fabian Chablais 1, Anna Jaźwińska 1
1Department of Biology, Unit of Zoology, University of Fribourg, Fribourg, Switzerland

Zebrafish represents a valuable model to study the mechanisms of heart regeneration in vertebrates. Here, we present a protocol for induction of a heart infarct in adult zebrafish using cryoinjury. This method results in massive cell death within 20% of the ventricular wall, similar to that observed in mammalian infarcts.

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Neuroscience

Appetitive Associative Olfactory Learning in Drosophila Larvae
Anthi A. Apostolopoulou 1, Annekathrin Widmann 1, Astrid Rohwedder 1,2, Johanna E. Pfitzenmaier 1, Andreas S. Thum 1,2
1Department of Biology, University of Konstanz, 2Department of Biology, University of Fribourg

Drosophila larvae are able to associate odor stimuli with gustatory reward. Here we describe a simple behavioral paradigm that allows the analysis of appetitive associative olfactory learning.

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Neuroscience

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae
Abud J. Farca Luna 1, Alina M. H. J. von Essen 1, Yves F. Widmer 1, Simon G. Sprecher 1
1Department of Biology, Institute of Cell and Developmental Biology, University of Fribourg

Here we describe a light-dark preference test for Drosophila larva. This assay provides information about innate and circadian regulation of light sensing and processing photobehavior.

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Environment

Optimize Flue Gas Settings to Promote Microalgae Growth in Photobioreactors via Computer Simulations
Lian He 1, Amelia B Chen 1,2, Yi Yu 2, Leah Kucera 3, Yinjie Tang 1
1Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis, 2School of Pharmaceutical Sciences, Wuhan University of China, 3Department of Earth and Planetary Sciences, Washington University in St. Louis

Flue gas from power plants is a cheap CO2 source for algal growth. We have built prototype "flue gas to algal cultivation" systems and described how to scale up the algal cultivation process. We have demonstrated the use of a mass-transfer bio-reaction model to simulate and to design the optimal operation of flue gas for the growth of Chlorella sp. in algal photo-bioreactors.

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Bioengineering

Cell-based Therapy for Heart Failure in Rat: Double Thoracotomy for Myocardial Infarction and Epicardial Implantation of Cells and Biomatrix
Aurélien Frobert 1, Jérémy Valentin 1, Stéphane Cook 1, Justine Lopes-Vicente 1, Marie-Noëlle Giraud 1
1Cardiology, Department of Medicine, University of Fribourg

Implantation of a biograft to treat myocardial infarction induced by LAD ligation in a rodent model has conventionally required two open-heart surgeries. In order to reduce mortality and provide optimal conditions for fixation of solid and gelatinous biomatrices associated with cells, minimally invasive procedures have been developed.

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Biology

Methylnitrosourea (MNU)-induced Retinal Degeneration and Regeneration in the Zebrafish: Histological and Functional Characteristics
Ellinor Maurer 1, Markus Tschopp 1,2, Christoph Tappeiner 1, Pauline Sallin 3, Anna Jazwinska 3, Volker Enzmann 1
1Department of Ophthalmology, Inselspital, University of Bern, 2Department of Ophthalmology, University Hospital of Basel, 3Department of Biology, University of Fribourg

Herein we demonstrate quantification of retinal de- and regeneration and its impact on visual function using N-methyl-N-nitrosourea in the adult zebrafish. Loss of visual acuity and decreased photoreceptor numbers were followed by proliferation in the inner nuclear layer. Complete morphological and functional regeneration occurred 30 days after the initial treatment.

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Neuroscience

Non-invasive Assessment of Changes in Corticomotoneuronal Transmission in Humans
Wolfgang Taube 1, Christian Leukel 1,2, Jens Bo Nielsen 3,4, Jesper Lundbye-Jensen 3,4
1Department of Medicine, Movement and Sport Science, University of Fribourg (Switzerland), 2Department of Sport Science, University of Freiburg (Germany), 3Department of Neuroscience and Pharmacology, University of Copenhagen, 4Department of Nutrition, Exercise and Sports, University of Copenhagen

The aim of the present study was to assess changes in transmission at the corticomotoneuronal synapses in humans after repetitive transcranial magnetic stimulation. For this purpose, an electrophysiological method is introduced that allows assessment of pathway specific corticospinal transmission, i.e. differentiation of fast, direct corticospinal pathways from polysynaptic connections.

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Biology

A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells
Farokh Dotiwala 1, Isabelle Fellay 2, Luis Filgueira 2, Denis Martinvalet 3, Judy Lieberman 1, Michael Walch 2
1Cellular and Molecular Medicine Program, Boston Children’s Hospital and Harvard Medical School, 2Department of Medicine, University of Fribourg, 3Centre Médical Universitaire, University of Geneva

We describe here a cost-efficient granzyme expression system using HEK293T cells that produces high yields of pure, fully glycosylated and enzymatically active protease.

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Behavior

Simple and Computer-assisted Olfactory Testing for Mice
Emanuele Brai 1, Lavinia Alberi 1
1Unit of Anatomy, Department of Medicine, University of Fribourg

We present a simple and unbiased olfactory test in mice. With this protocol olfactory discrimination, preference, avoidance and sensitivity to a novel odor as compared to water can be assessed in single behavioral sessions. This method is indicated for a single experimenter and analysis is based on computer-assisted video processing.

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Behavior

Force and Position Control in Humans - The Role of Augmented Feedback
Benedikt Lauber 1,2, Martin Keller 2, Christian Leukel 1,3, Albert Gollhofer 1, Wolfgang Taube 2
1Department of Sport Science, University of Freiburg, 2Department of Medicine, Movement and Sport Science, University of Fribourg, 3Bernsteincenter Freiburg

Controlling an identical movement with position or force feedback results in different neural activation and motor behavior. This protocol describes how to investigate behavioral changes by looking at neuromuscular fatigue and how to evaluate motor cortical (inhibitory) activity using subthreshold TMS with respect to the interpretation of augmented feedback.

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Biology

En Face Detection of Nitric Oxide and Superoxide in Endothelial Layer of Intact Arteries
Yi Yu 1,2, Yuyan Xiong 1,2, Jean-Pierre Montani 2,3, Zhihong Yang 1,2, Xiu-Fen Ming 1,2
1Cardiovascular and Aging Research, Department of Medicine, Division of Physiology, Faculty of Science, University of Fribourg, 2Kidney Control of Homeostasis, Swiss National Centre of Competence in Research, 3System Physiology, Department of Medicine, Division of Physiology, Faculty of Science, University of Fribourg

In this article we describe an adapted relatively easy method using the fluorescence dye diaminofluorescein-2 diacetate (DAF-2DA) and dihydroethidium (DHE) for en face simultaneous detection and visualization of intracellular nitric oxide (NO) and superoxide anion (O2.−) respectively, in freshly isolated intact aortas of an obesity mouse model.

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Medicine

Histological Quantification of Chronic Myocardial Infarct in Rats
Jérémy Valentin 1, Aurélien Frobert 1, Guillaume Ajalbert 1, Stéphane Cook 1, Marie-Noëlle Giraud 1
1Cardiology, Department of Medicine, University of Fribourg

The post-mortem assessment of myocardial infarction (MI) in rodents is based on quantification of the infarct on stained heart sections. We describe an accurate method to quantify the infarct size using systematic sampling of harvested rat hearts from base to apex and image analyses of trichrome-stained histological sections.

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Medicine

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury
Manish Jain *1, Aurélien Frobert *1, Jérémy Valentin 1, Stéphane Cook 1, Marie-Noëlle Giraud 1
1Cardiology, Department of Medicine, University of Fribourg

Animal models of atherosclerosis are essential to understand the mechanism and to investigate newer approaches to prevent plaque development or rupture, a leading cause of death in the industrialized world. This protocol uses a combination of balloon injury and cholesterol rich diet to induce atherosclerotic plaques in rabbit iliac artery.

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Behavior

Intracortical Inhibition Within the Primary Motor Cortex Can Be Modulated by Changing the Focus of Attention
Yves-Alain Kuhn 1, Martin Keller 1, Jan Ruffieux 1, Wolfgang Taube 1
1Department of Medicine, Movement and Sport Sciences, University of Fribourg

Using two different transcranial magnetic stimulation (TMS) protocols, this manuscript describes how to measure and compare cortical inhibition within the primary motor cortex when adopting different attentional foci.

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Immunology and Infection

Evaluation of Extracellular Vesicle Function During Malaria Infection
María Andrea Hernández-Castañeda *1, Smart Mbagwu *1, Kehinde Adebayo Babatunde *1, Michael Walch 1, Luis Filgueira 1, Pierre-Yves Mantel 1
1Department of Medicine, University of Fribourg

In this work, we describe protocols to investigate the role of extracellular vesicles (EVs) released by Plasmodium falciparum infected erythrocytes. In particular, we focus on the interactions of EVs with endothelial cells.

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Neuroscience

An Alternative Approach to Study Primary Events in Neurodegeneration Using Ex Vivo Rat Brain Slices
Emanuele Brai 1, Antonella Cogoni 1, Susan A. Greenfield 1
1Neuro-Bio Ltd, Building F5, Culham Science Centre

We present a method which can provide further insights into the early events underlying neurodegeneration and based on the established ex-vivo brain technique, combining the advantages of in vivo and in vitro experiments. Moreover, it represents a unique opportunity for direct comparison of treated and untreated group in the same anatomical plane.

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Biology

Intrathoracic Injection for the Study of Adult Zebrafish Heart
Thomas Bise 1, Anna Jaźwińska 1
1Department of Biology, University of Fribourg

This method relies on the injection of 0.5−3 μL of solution into the thorax of adult zebrafish. The procedure efficiently delivers proteins and chemical compounds into the proximity of the zebrafish heart without damaging the organ. The approach is suitable for testing effects of exogenous factors on various tissues of the heart.

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JoVE Core

Multicellular Human Alveolar Model Composed of Epithelial Cells and Primary Immune Cells for Hazard Assessment
Hana Barosova *1, Barbara Drasler *1, Alke Petri-Fink 1, Barbara Rothen-Rutishauser 1
1Adolphe Merkle Institute, University of Fribourg

Presented here is a protocol for primary human blood monocyte isolation as well as their differentiation into macrophages and dendritic cells and assembly with epithelial cells into a multicellular human lung model. Biological responses of cocultures composed of immune cells differentiated from either freshly isolated or thawed monocytes, upon exposure to proinflammatory stimuli, are compared.

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Environment

Investigating the Relationship between Sea Surface Chlorophyll and Major Features of the South China Sea with Satellite Information
Huan-Huan Chen *1,2, Rui Tang *2, Hao-Ran Zhang *1,2, Yi Yu 2, Yuntao Wang 2
1College of Oceanography, Hohai University, 2State Key Laboratory of Satellite Ocean Environment Dynamics, Second Institute of Oceanography, Ministry of Natural Resources

Sea surface chlorophyll, temperature, sea level height, wind, and front data obtained or derived from satellite observations offer an effective way to characterize the ocean. Presented is a method for the comprehensive study of these data, including overall average, seasonal cycle, and intercorrelation analyses, to fully understand regional dynamics and ecosystems.

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JoVE Core

Cultivating a Three-dimensional Reconstructed Human Epidermis at a Large Scale
Irini M. Dijkhoff 1, Benedetta Petracca 1,2, Roxane Prieux 3, Giuseppe Valacchi 3,4, Barbara Rothen-Rutishauser 1, Marc Eeman 2
1Adolphe Merkle Institute, University of Fribourg, 2Dow Silicones Belgium SRL, 3Department of Biomedical and Specialist Surgical Sciences, University of Ferrara, 4Department of Animal Sciences, Plants for Human Health Institute, North Carolina State University

This protocol describes a straightforward method to cultivate three-dimensional reconstituted human epidermis in a reproducible and robust manner. Additionally, it characterizes the structure-function relationship of the epidermal barrier model. The biological responses of the reconstituted human epidermis upon proinflammatory stimuli are also presented.

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Biology

A Cryoinjury Model for Studying Skeletal Muscle Regeneration of the Caudal Peduncle in Adult Zebrafish
Hendrik Oudhoff 1, Florian Baumgartner 1, Anna Jaźwińska 1
1Department of Biology, University of Fribourg

This protocol describes a cryoinjury model to induce profound damage of several caudal myomeres in adult zebrafish. This method provides a new approach for studying skeletal muscle regeneration after severe loss of tissue in non-mammalian vertebrates.

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