Temple University School of Medicine

5 ARTICLES PUBLISHED IN JoVE

Neuroscience

Subcutaneous Administration of Muscarinic Antagonists and Triple-Immunostaining of the Levator Auris Longus Muscle in Mice

Megan Wright¹, Amy Kim², Young-Jin Son³

¹Biology Department, Arcadia University, ²Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, ³Shriners Hospitals Pediatric Research Center and Department of Anatomy and Cell Biology, Temple University School of Medicine

We describe procedures for repeated administration of inhibitors of muscarinic signaling to the levator auris longus (LAL) muscle of young adult mice and for subsequent immunostaining of its neuromuscular junctions (NMJs) in wholemounts. The LAL muscle has unique advantages for revealing in vivo pharmacological effects on NMJs.

Neuroscience

Live Imaging of Dorsal Root Axons after Rhizotomy

Andrew Skuba¹, B. Timothy Himes^2,3, Young-Jin Son⁴

¹Temple University, Shriners Hospitals Pediatric Research Center and Department of Anatomy and Cell Biology, ²Medical Research Service, Department of Veterans Affairs Hospital, ³Department of Neurobiology and Anatomy, Drexel University College of Medicine, ⁴Shriners Hospitals Pediatric Research Center and Department of Anatomy and Cell Biology, Temple University School of Medicine

An in vivo imaging protocol to monitor primary sensory axons following dorsal root crush is described. The procedures utilize wide-field fluorescence microscopy and thy1-YFP transgenic mice, and permit repeated imaging of axon regeneration over 4 cm in the PNS and axon interactions with the interface of the CNS.

Neuroscience

Complete Spinal Cord Injury and Brain Dissection Protocol for Subsequent Wholemount In Situ Hybridization in Larval Sea Lamprey

Antón Barreiro-Iglesias¹, Guixin Zhang², Michael E. Selzer^2,3, Michael I. Shifman²

¹Centre for Neuroregeneration, School of Biomedical Sciences, University of Edinburgh, ²Shriners Hospitals Pediatric Research Center (Center for Neural Repair and Rehabilitation), Temple University School of Medicine, ³Department of Neurology, Temple University School of Medicine

Lampreys recover locomotion after a complete spinal cord injury. However, some spinal-projecting neurons are good regenerators and others are not. This paper illustrates the techniques for housing sea lamprey larvae (and recently transformed adults), producing complete spinal cord transections and preparing wholemount brains and spinal cords for in situ hybridization.

Developmental Biology

A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome

Ping Xue¹, Zhiqiang Wang¹, Xiaomin Fu^1,2, Junjiang Wang^1,3, Gopika Punchhi¹, Andrew Wolfe^1,4, Sheng Wu^1,4,5

¹Department of Pediatrics, Johns Hopkins University School of Medicine, ²Department of Health, Beijing Military General Hospital, ³Southern Medical University, ⁴Department of Molecular and Cellular Physiology, Johns Hopkins University School of Medicine, ⁵Department of Gynecology and Obstetrics, Johns Hopkins University School of Medicine

We describe the development of a lean PCOS-like mouse model with dihydrotestosterone pellet to study the pathophysiology of PCOS and the offspring from these PCOS-like dams.

Biology

An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation

Mingxiao Feng¹, Sara Divall³, Sheng Wu^1,2

¹Department of Pediatrics, Johns Hopkins University School of Medicine, ²Department of Physiology, Temple University School of Medicine, ³Department of Pediatrics, Seattle Children’s Hospital

Primary hepatocytes are a valuable tool to study liver response and metabolism in vitro. Utilizing commercially available reagents, an improved time- and labor-efficient protocol for mouse primary hepatocyte isolation was developed.