Shiraz University of Medical Sciences View Institution's Website 11 articles published in JoVE Biochemistry Rapid and Cost-Effective RNA Extraction of Rat Pancreatic Tissue Sanaz Dastghaib1,2, Zahra Shahsavar3, Zahra Karimian4, Pooneh Mokarram1,5 1Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, 2Endocrinology and Metabolism Research Center, Nemazee Hospital, Shiraz University of Medical Sciences, 3English Language Department, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, 4Department of e-Learning in Medical Sciences, Virtual School and Center of Excellence in e-Learning, Shiraz University of Medical Sciences, 5Autophagy Research Center, School of Medicine, Shiraz University of Medical Sciences The purity and integrity of the isolated RNA is a vital step in RNA dependent assays. Here, we present a practical, rapid, and inexpensive method to extract RNA from a small quantity of undamaged pancreatic tissue. Biochemistry Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay Javad Alizadeh1,2, Shahla Shojaei1,2,3, Simone da Silva Rosa1, Adel Rezaei Moghadam1,2, Amir A. Zeki4, Mohammad Hashemi5, Marek J. Los6,7,8, Joseph W. Gordon1,9, Saeid Ghavami1,2,10 1Department of Human Anatomy and Cell Science, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, 2Biology of Breathing Theme, Children Hospital Research Institute of Manitoba, University of Manitoba, 3Department of Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, 4Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, Center for Comparative Respiratory Biology and Medicine, 5Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, 6Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, 7Centre de biophysique moléculaire - UPR 4301, Centre national de la recherche scientifique (CNRS) CS80054, 8LinkoCare Life Sciences AB, 9College of Nursing and Children's Hospital Research Institute of Manitoba, Rady Faculty of Health Sciences, University of Manitoba, 10Health Policy Research Center, Institute of Health, Shiraz University of Medical Sciences Here we describe a protocol to investigate the prenylation and guanosine-5'-triphosphate (GTP)-loading of Rho GTPase. This protocol consists of two detailed methods, namely membrane fractionation and a GTPase-linked immunosorbent assay. The protocol can be used for measuring the prenylation and GTP loading of different other small GTPases. Medicine Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion Robert W. Regenhardt1, Saeed Ansari2, Hassan Azari3, Kenneth J. Caldwell2, Adam P. Mecca1 1Department of Physiology and Functional Genomics, University of Florida, 2Department of Neurosurgery, McKnight Brain Institute, University of Florida, 3Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo. Medicine Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat Saeed Ansari1, Hassan Azari2, Kenneth J. Caldwell1, Robert W. Regenhardt3, Vishnumurthy S. Hedna4, Micheal F. Waters4, Brian L. Hoh1, Adam P. Mecca3 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran, 3Department of Physiology and Functional Genomics, University of Florida, 4Department of Neurology, University of Florida Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed. Medicine Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE) Loic P. Deleyrolle1, Mark R. Rohaus1, Jeff M. Fortin1, Brent A. Reynolds1, Hassan Azari1,2 1Department of Neurosurgery, The University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran This video protocol demonstrates the application of the fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) for the identification and separation of different sub-populations of cells in human glioblastoma based on frequency of cell division. Neuroscience The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytometry Hassan Azari1,2, Sharareh Sharififar1, Jeff M. Fortin1, Brent A. Reynolds1 1Department of Neurosurgery, The University of Florida, 2Laboratory for Stem Cell Research, Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran This video protocol demonstrates a novel method for the generation and subsequent purification of neuronal progenitor cells from a renewable source of neural stem cells (NSCs) based on their physical (size and internal granularity) and fluorescent properties using flow cytometry technology. Neuroscience Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay Hassan Azari1,2, Sebastien Millette1, Saeed Ansari1, Maryam Rahman1, Loic P. Deleyrolle1, Brent A. Reynolds1 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences This video protocol demonstrates the isolation and expansion of stem like cells from surgically resected human glioblastoma mutliforme (GBM) tumor tissue using the neurosphere assay culture method. Medicine Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice Saeed Ansari*1, Hassan Azari*1,2, Douglas J. McConnell1, Aqeela Afzal1, J Mocco1 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results. Neuroscience Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells Hassan Azari1,2, Sharon A. Louis3, Sharareh Sharififar1, Vinata Vedam-Mai1, Brent A. Reynolds1 1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, 3STEMCELL Technologies, Inc. This video protocol demonstrates how to discriminate and enumerate bona fide neural stem cells in a mixed population of neural precursor cells using the neural colony-forming cell assay. Neuroscience Establishing Embryonic Mouse Neural Stem Cell Culture Using the Neurosphere Assay Hassan Azari1,2, Sharareh Sharififar2, Maryam Rahman2, Saeed Ansari2, Brent A. Reynolds2 1Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran, 2Department of Neurosurgery, The University of Florida This video protocol demonstrates the application of the neurosphere assay for the isolation and expansion of neural stem cells from the ganglionic eminences of embryonic day 14-mouse brain. Neuroscience Isolation and Expansion of the Adult Mouse Neural Stem Cells Using the Neurosphere Assay Hassan Azari1,2, Maryam Rahman2, Sharareh Sharififar2, Brent A. Reynolds2 1 Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran, 2Department of Neurosurgery, University of Florida This video protocol demonstrates the neurosphere assay method to generate and expand neural stem cells from the adult mouse periventricular region, and provides technical insights to ensure one can achieve reproducible neurosphere cultures.
