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SUNY Upstate Medical University

12 ARTICLES PUBLISHED IN JoVE

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Biology

Tissue Determination Using the Animal Cap Transplant (ACT) Assay in Xenopus laevis
Andrea S. Viczian 1, Michael E. Zuber 1
1Department of Ophthalmology, SUNY Upstate Medical University

Animal caps overexpressing gene product(s) are transplanted to the flank of developing Xenopus laevis embryos in order to establish whether tissue is determined.

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Medicine

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage
Rahul V. Dudhani 1, Michele Kyle 1, Christina Dedeo 2, Margaret Riordan 1, Eric M. Deshaies 1,2
1Department of Neurosurgery, SUNY Upstate Medical University, 2Department of Neuroscience and Physiology, SUNY Upstate Medical University

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

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Immunology and Infection

A Quantitative Evaluation of Cell Migration by the Phagokinetic Track Motility Assay
Maciej T. Nogalski 1,2, Gary C.T. Chan 3, Emily V. Stevenson 1,2, Donna K. Collins-McMillen 1,2, Andrew D. Yurochko 1,2,4
1Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, 2Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, 3Department of Microbiology and Immunology, SUNY Upstate Medical University, 4Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center

The phagokinetic motility track assay is a method used to assess the movement of cells. Specifically, the assay measures chemokinesis (random cell motility) over time in a quantitative manner. The assay takes advantage of the ability of cells to create a measurable track of their movement on colloidal gold-coated coverslips.

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Biology

Measurement of Vacuolar and Cytosolic pH In Vivo in Yeast Cell Suspensions
Theodore T. Diakov 1, Maureen Tarsio 1, Patricia M. Kane 1
1Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University

Vacuolar and cytosolic pH can be measured in live yeast (S. cerevisiae) cells using ratiometric fluorescent dyes localized to specific cellular compartments. We describe procedures for measuring vacuolar pH with BCECF-AM, which localizes to the vacuole in yeast, and cytosolic pH with a cytosolic ratiometric pH-sensitive GFP (yeast pHluorin).

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Neuroscience

Ex utero Electroporation and Whole Hemisphere Explants: A Simple Experimental Method for Studies of Early Cortical Development
Anna J. Nichols *1, Ryan S. O'Dell *1, Teresa A. Powrozek *1, Eric C. Olson 1
1Department of Neuroscience and Physiology, SUNY Upstate Medical University

This protocol describes an improved explant procedure that involves ex utero electroporation, dissection and culture of entire cerebral hemispheres from the embryonic mouse. The preparation facilitates pharmacological studies and assays of gene function during early cortical development.

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Chemistry

Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
Naman B. Shah 1, Thomas M. Duncan 1
1Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University

The protocols here describe kinetic assays of protein-protein interactions with Bio-layer Interferometry. F-type ATP synthase, which is involved in cellular energy metabolism, can be inhibited by its ε subunit in bacteria. We have adapted Bio-layer Interferometry to study interactions of the catalytic complex with ε’s inhibitory C-terminal domain.

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Medicine

Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms
Eric M Deshaies 1,2, Mark R Villwock 1, Amit Singla 3, Gentian Toshkezi 4, David J Padalino 1
1Neurovascular and Stroke Center, Crouse Neuroscience Institute, 2Department of Biology, Syracuse University, 3Department of Neurosurgery, University of Florida, 4Department of Neurosurgery, SUNY Upstate Medical University

Minimally invasive thumb-sized pterional craniotomy for aneurysm clipping has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

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Neuroscience

A Simple Behavioral Assay for Testing Visual Function in Xenopus laevis
Andrea S. Viczian 1, Michael E. Zuber 1
1Ophthalmology Department, Center for Vision Research, SUNY Eye Institute, Upstate Medical University

Xenopus laevis tadpoles prefer swimming on the white side of a black/white tank. This behavior is guided by their vision. Based on this behavior, we present a simple assay to test the visual function of tadpoles.

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Biology

A Simplified System for Evaluating Cell Mechanosensing and Durotaxis In Vitro
Gregory J. Goreczny 1, Duncan B. Wormer 1, Christopher E. Turner 1
1Department of Cell & Developmental Biology, SUNY Upstate Medical University

Many mammalian cells preferentially migrate towards a more rigid matrix or substrate through durotaxis. The goal of this protocol is to provide a simple in vitro system that can be used to study and manipulate cell durotaxis behaviors by incorporating polydimethylsiloxane (PDMS) substrates of defined rigidity, interfacing with glass coverslips.

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Cancer Research

Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models
Disharee Nath 1,2, Julie R. White 3,4, Gennady Bratslavsky 1, Leszek Kotula 1,2
1Department of Urology, SUNY Upstate Medical University, 2Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 3Laboratory of Comparative Pathology, Memorial Sloan-Kettering Cancer Center, 4Boulder BioPATH, Inc

Genetically engineered mice are useful models for investigating prostate cancer mechanisms. Here we present a protocol to identify and dissect prostate lobes from a mouse urogenital system, differentiate them based on histology, and isolate and culture the primary prostate cells in vitro as spheroids for downstream analyses.

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Medicine

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies
Nileyma Castro *1, Stephanie R. Gillespie *2, Audrey M. Bernstein 1
1Department of Ophthalmology, SUNY Upstate Medical University, 2Department of Dermatology, Columbia University Medical Center

A protocol for an ex vivo corneal organ culture model useful for wound healing studies is described. This model system can be used to assess the effects of agents to promote regenerative healing or drug toxicity in an organized 3D multicellular environment.

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Neuroscience

The Impact of Motor Task Conditions on Goal-Directed Arm Reaching Kinematics and Trunk Compensation in Chronic Stroke Survivors
Jaimie Girnis 1,2, Tarek Agag 1, Tobias Nobiling 1,3, Vanessa Sweet 1,4, Bokkyu Kim 1
1Department of Physical Therapy Education, SUNY Upstate Medical University, 2College of Health and Rehabilitation Sciences: Sargent College Center for Neurorehabilitation, Boston University, 3Department of Physical Medicine and Rehabilitation, University of Rochester Medical Center, 4Rehabilitation Today

This protocol is intended to investigate the impact of task conditions on movement strategies in chronic stroke survivors. Further, this protocol can be used to examine if a restriction in elbow extension induced by neuromuscular electrical stimulation causes trunk compensation during goal-directed arm reaches in non-disabled adults.

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