SUNY Upstate Medical University View Institution's Website 14 articles published in JoVE Neuroscience A Gravity-Fed Transcardial Perfusion Method for Histologic Analysis of the Mouse Central Nervous System Arnav Rana1, Paul T. Massa2, Xin Jie Chen1,3 1Department of Biochemistry and Molecular Biology, State University of New York Upstate Medical University, 2Department of Neurology, State University of New York Upstate Medical University, 3Department of Neuroscience and Physiology, State University of New York Upstate Medical University A convenient gravity-fed perfusion method for histological analysis of the mouse central nervous system is presented. Immunofluorescent detection of phosphorylated α-synuclein is demonstrated in a mouse model of Parkinson's disease. This work also comprehensively describes the transcardial perfusion, dissection, tissue freezing/embedding, and frozen sectioning steps. Neuroscience The Impact of Motor Task Conditions on Goal-Directed Arm Reaching Kinematics and Trunk Compensation in Chronic Stroke Survivors Jaimie Girnis1,2, Tarek Agag1, Tobias Nobiling1,3, Vanessa Sweet1,4, Bokkyu Kim1 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. Medicine Multi-system Monitoring for Identification of Seizures, Arrhythmias and Apnea in Conscious Restrained Rabbits Cameron Bosinski1, Kyle Wagner1, Xiangping Zhou2, Lynn Liu3, David S. Auerbach1 1Department of Pharmacology, Upstate Medical University, 2Department of Neurology, Upstate Medical University, 3Strong Epilepsy Center, Departments of Neurology, Anesthesiology / Perioperative Medicine, & Pediatrics, University of Rochester Medical Center Using simultaneous video-EEG-ECG-oximetry-capnography, we developed a methodology to evaluate the susceptibility of rabbit models to develop provoked arrhythmias and seizures. This novel recording system establishes a platform to test the efficacy and safety of therapeutics and can capture the complex cascade of multi-system events that culminate in sudden death. Medicine Ex Vivo Corneal Organ Culture Model for Wound Healing Studies Nileyma Castro*1, Stephanie R. Gillespie*2, Audrey M. Bernstein1 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. Cancer Research Identification, Histological Characterization, and Dissection of Mouse Prostate Lobes for In Vitro 3D Spheroid Culture Models Disharee Nath1,2, Julie R. White3,4, Gennady Bratslavsky1, Leszek Kotula1,2 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. Biology A Simplified System for Evaluating Cell Mechanosensing and Durotaxis In Vitro Gregory J. Goreczny1, Duncan B. Wormer1, Christopher E. Turner1 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. Medicine Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms Eric M Deshaies1,2, Mark R Villwock1, Amit Singla3, Gentian Toshkezi4, David J Padalino1 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. Chemistry Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects Naman B. Shah1, Thomas M. Duncan1 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. Biology Measurement of Vacuolar and Cytosolic pH In Vivo in Yeast Cell Suspensions Theodore T. Diakov1, Maureen Tarsio1, Patricia M. Kane1 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). 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. Olson1 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. Biology Analysis of Gene Function and Visualization of Cilia-Generated Fluid Flow in Kupffer's Vesicle Guangliang Wang1, H. Joseph Yost2, Jeffrey D. Amack1 1Department of Cell and Developmental Biology, State University of New York, Upstate Medical University, 2Department of Neurobiology and Anatomy, Eccles Institute of Human Genetics, University of Utah Cilia-generated fluid flow in Kupffer’s Vesicle (KV) controls left-right patterning of the zebrafish embryo. Here, we describe a technique to modulate gene function specifically in KV cells. In addition, we show how to deliver fluorescent beads into KV to visualize fluid flow. Medicine A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage Rahul V. Dudhani1, Michele Kyle1, Christina Dedeo2, Margaret Riordan1, Eric M. Deshaies1,2 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. Immunology and Infection A Quantitative Evaluation of Cell Migration by the Phagokinetic Track Motility Assay Maciej T. Nogalski1,2, Gary C.T. Chan3, Emily V. Stevenson1,2, Donna K. Collins-McMillen1,2, Andrew D. Yurochko1,2,4 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. Biology Tissue Determination Using the Animal Cap Transplant (ACT) Assay in Xenopus laevis Andrea S. Viczian1, Michael E. Zuber1 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.
