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university of kentucky

14 ARTICLES PUBLISHED IN JoVE

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Biology

Measures of Heart and Ventilatory Rates in Freely Moving Crayfish
Sonya M. Bierbower 1, Robin L. Cooper 1
1Department of Biology, University of Kentucky

Invertebrates show an autonomic sympathetic-like response similar to that described for vertebrates. The coordination of the cardio-vascular and ventilatory systems allows for measurement of a biological index in which to assess an organism internal state.

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Biology

Historical View and Physiology Demonstration at the NMJ of the Crayfish Opener Muscle
Ann S. Cooper 1, Robin L. Cooper 1
1Department of Biology, University of Kentucky

The opener muscle of the crayfish leg is presented for its historical importance and experimental versatility in muscle phenotype, synaptic physiology and plasticity.

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Biology

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies
Ann S. Cooper 1, Kylah E. Rymond 1, Matthew A. Ward 1, Easter L. Bocook 1, Robin L. Cooper 1
1Department of Biology, University of Kentucky, Lexington

We present various ways to monitor heart function in the larva of Drosophila for assessing questions dealing with the function of gap junctions, ion channel mutations, modulation of pacemaker activity and pharmacological studies.

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Neuroscience

Physiological Recordings of High and Low Output NMJs on the Crayfish Leg Extensor Muscle
Wen Hui Wu *1, Robin L. Cooper *1
1Department of Biology, University of Kentucky

This article demonstrates how to conduct electrophysiological recordings of synaptic responses on the extensor muscle in the walking leg of a crayfish and how the nerve terminals are visualized to show the gross morphological differences of high- and low-output nerve terminals.

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Neuroscience

Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises
Brittany Baierlein *1, Alison L. Thurow *1, Harold L. Atwood *2, Robin L. Cooper *1
1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto

The experiments demonstrate an easy approach for students to gain experience in examining muscle structure, synaptic responses, the effects of ion gradients and permeability on membrane potentials. Also, a sensory-CNS-motor-muscle circuit is presented to show a means to test effects of compounds on a neuronal circuit.

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Neuroscience

Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception
Bonnie Leksrisawat *1, Ann S. Cooper *1, Allison B. Gilberts *1, Robin L. Cooper *1
1Department of Biology, University of Kentucky

The primary purpose of this experiment is to understand how primary sensory neurons convey information of joint movements and positions as proprioceptive information for an animal. An additional objective of this report is present the anatomy of the preparation by dissection and viewing of neurons under a dissecting microscope.

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Neuroscience

Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise
Ann S. Cooper *1, Bonnie Leksrisawat *1, Allison B. Gilberts *1, A. Joffre Mercier *2, Robin L. Cooper *1
1Department of Biology, University of Kentucky, 2Department of Biological Sciences, Brock University

In this report we demonstrate techniques that can be used to investigate the biology of the crayfish hindgut. We show how to dissect a crayfish abdomen and study the associated anatomy, physiology and modulation of activity. The peristaltic activity and strength of contractions are measured using a force transducer.

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Education

Modeling Biological Membranes with Circuit Boards and Measuring Electrical Signals in Axons: Student Laboratory Exercises
Martha M. Robinson 1, Jonathan M. Martin 1, Harold L. Atwood 2, Robin L. Cooper 1
1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto

This is a demonstration of how biological membranes can be understood using electrical models. We also demonstrate procedures for recording action potentials from the ventral nerve cord of the crayfish for student orientated laboratories.

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Immunology and Infection

Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells
Gavin I. Ellis 1, Mary Catherine Reneer 1, Alejandra Catalina Vélez-Ortega 1, Andrea McCool 1, Francesc Martí 1
1Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky

We describe a method for generating regulatory, memory and naïve T cells from a single human blood donor. Polarized Tregs can be then compared to other subsets in a variety of genetic and functional applications with genetic homogeneity, including a suppression assay also detailed here.

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Immunology and Infection

Oral Transmission of Listeria monocytogenes in Mice via Ingestion of Contaminated Food
Elsa N. Bou Ghanem 1, Tanya Myers-Morales 1, Grant S. Jones 1, Sarah E.F. D'Orazio 1
1Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky

This paper describes a novel method for oral infection of mice using Listeria monocytogenes-contaminated food. The protocol can readily be adapted for use with other food borne bacterial pathogens.

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Neuroscience

Neural Circuit Recording from an Intact Cockroach Nervous System
Josh S. Titlow 1, Zana R. Majeed 1,2, H. Bernard Hartman 3, Ellen Burns 1, Robin L. Cooper 1
1Department of Biology, University of Kentucky , 2Department of Biology, University of Salahaddin, 3Oregon Institute of Marine Biology, University of Oregon

This article describes the cockroach ventral nerve cord dissection and extracellular recordings from the cercal nerve and connectives. Evoked responses are generated by electrical stimulation of the cercal nerve or direct mechanical stimulation of the cerci.

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Neuroscience

Intracellular Recording, Sensory Field Mapping, and Culturing Identified Neurons in the Leech, Hirudo medicinalis
Josh Titlow 1, Zana R. Majeed 1,2, John G Nicholls 3, Robin L. Cooper 1
1Department of Biology, University of Kentucky, 2Department of Biology, College of Science, University of Salahaddin, Iraq, 3Department of Neurobiology and Cognitive Neuroscience, SISSA, Italy

This article describes three nervous system preparations using leeches: intracellular recording from neurons in ventral ganglia, culturing neurons from ventral ganglia, and recording from a patch of innervated skin to map sensory fields.

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Biology

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
Rekha Kushwaha 1, Kim R. Schäfermeyer 1, A. Bruce Downie 1
1Department of Horticulture, University of Kentucky

Phage display is a powerful technique to capture proteins or protein moieties that interact with an immobilized molecule of interest. Once a decision of the type of phage cDNA library to create and screen has been made, the protocol described here permits efficient affinity selection leading to identification of interactors.

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JoVE Journal

Proprioception and Tension Receptors in Crab Limbs: Student Laboratory Exercises
Zana R. Majeed 1,2, Josh Titlow 1,2, H. Bernard Hartman 3, Robin Cooper 1,2
1Department of Biology, University of Kentucky, 2Center of Muscle Biology, University of Kentucky, 3Oregon Institute of Marine Biology, University of Oregon

Physiological and anatomical techniques are demonstrated to address function and structure for joint proprioceptors and muscle tension receptors in crustacean walking limbs.

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