Indiana University-Purdue University Indianapolis

6 ARTICLES PUBLISHED IN JoVE

JoVE Core

Local Application of Drugs to Study Nicotinic Acetylcholine Receptor Function in Mouse Brain Slices

Staci E. Engle¹, Hilary J. Broderick¹, Ryan M. Drenan¹

¹Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University

In this paper, we describe a useful method to study ligand-gated ion channel function in neurons of acutely isolated brain slices. This method involves the use of a drug-filled micropipette for local application of drugs to neurons recorded using standard patch clamp techniques.

Chemistry

Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry

Elijah N. McCool^*1, Rachele Lubeckyj^*1, Xiaojing Shen¹, Qiang Kou², Xiaowen Liu^2,3, Liangliang Sun¹

¹Department of Chemistry, Michigan State University, ²Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, ³Center for Computational Biology and Bioinformatics, Indiana University School of Medicine

A detailed protocol is described for the separation, identification, and characterization of proteoforms in protein samples using capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS). The protocol can be used for the high-resolution characterization of proteoforms in simple protein samples and the large-scale identification of proteoforms in complex proteome samples.

Behavior

The Participant-Reported Implementation Update and Score (PRIUS): A Novel Method for Capturing Implementation-Related Data Over Time

Edward J. Miech^1,2,3,4,5, Nicholas A. Rattray^1,2,3,6, Dawn M. Bravata^1,2,3,5, Jennifer Myers^1,3, Teresa M. Damush^1,2,3,5

¹VA Precision Monitoring (PRIS-M) QUERI, Richard L. Roudebush VA Medical Center, ²William M. Tierney Center for Health Services Research, Regenstrief Institute, ³VA Health Services Research & Development Center for Health Information and Communication (CHIC), Richard L. Roudebush VA Medical Center, ⁴Department of Emergency Medicine, Indiana University School of Medicine, ⁵Department of Internal Medicine and Geriatrics, Indiana University School of Medicine, ⁶Department of Anthropology, Indiana University-Purdue University Indianapolis

This protocol describes a novel method for collecting and analyzing data related to ongoing implementation called the Participant-Reported Implementation Update and Score (PRIUS). The PRIUS method allows for the efficient and systematic capture of data over time and from multiple viewpoints in healthcare settings.

Biology

Artificial Intelligence Approaches to Assessing Primary Cilia

Ruchi Bansal¹, Staci E. Engle¹, Tisianna K. Kamba¹, Kathryn M. Brewer¹, Wesley R. Lewis², Nicolas F. Berbari^1,3,4

¹Department of Biology, Indiana University-Purdue University Indianapolis, ²Nikon Instruments Inc., ³Stark Neurosciences Research Institute, Indiana University, ⁴Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine

The use of artificial intelligence (Ai) to analyze images is emerging as a powerful, less biased, and rapid approach compared with commonly used methods. Here we trained Ai to recognize a cellular organelle, primary cilia, and analyze properties such as length and staining intensity in a rigorous and reproducible manner.

Medicine

A Reproducible Cartilage Impact Model to Generate Post-Traumatic Osteoarthritis in the Rabbit

Julian Dilley^*1,2,3, Hessam Noori-Dokht^*3,4,6, Abhijit Seetharam¹, Margaret Bello^1,3, Aaron Nanavaty^1,3, Roman M. Natoli^2,3, Todd McKinley^1,2,3, Zachary Bault⁵, Diane Wagner^2,3,6, Uma Sankar^1,3

¹Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, ²Department of Orthopaedic Surgery, Indiana University School of Medicine, ³Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, ⁴School of Mechanical Engineering, Purdue University, ⁵Large Animal Resource Center, Indiana University School of Medicine, ⁶Department of Mechanical and Energy Engineering, Indiana University-Purdue University Indianapolis

The open medial femoral condyle impact model in rabbits is reliable for studying post-traumatic osteoarthritis (PTOA) and novel therapeutic strategies to mitigate PTOA progression. This protocol generates an isolated cartilage defect of the posterior medial femoral condyle in rabbits using a carriage-based drop tower with an impactor head.

Bioengineering

Practical Considerations for the Design, Execution, and Interpretation of Studies Involving Whole-Bone Bending Tests of Rodent Bones

Olivia N. Reul¹, Ashtin M. Anneken¹, Rachel K. Kohler¹, Dyann M. Segvich¹, Joseph M. Wallace¹

¹Department of Biomedical Engineering, Indiana University-Purdue University Indianapolis

Mechanical testing of rodent bones is a valuable method for extracting information regarding a bone's susceptibility to fracture. Lacking proper practical understanding, the results may be overinterpreted or lack validity. This protocol will serve as a guide to ensure mechanical tests are performed accurately to provide valid and functional data.