Oakland University

9 ARTICLES PUBLISHED IN JoVE

Biology

RNAi Interference by dsRNA Injection into Drosophila Embryos

Ekaterini Iordanou¹, Rachana R. Chandran¹, Nicholas Blackstone¹, Lan Jiang¹

¹Department of Biological Sciences, Oakland University

RNA interference has been proven very effective to analyze gene function in Drosophila tracheal development. A detailed protocol used by Jiang lab to inject dsRNA into fly embryos to knockdown gene expression is illustrated. This technique has the potential for screening genes required for tissue and organ development in Drosophila.

Medicine

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Li Zuo^1,2, William J. Roberts^1,2, Kevin D. Evans¹

¹Radiologic Sciences & Respiratory Therapy Division, School of Health and Rehabilitation Sciences, Davis Heart and Lung Research Institute, The Ohio State University College of Medicine, ²Department of Biological Sciences, Oakland University

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

Developmental Biology

Isolation and Characterization of Mesenchymal Stromal Cells from Human Umbilical Cord and Fetal Placenta

Naimisha Beeravolu^1,2, Christina McKee^1,2, Ali Alamri^1,2, Sasha Mikhael³, Christina Brown^1,2, Mick Perez-Cruet^2,4, G. Rasul Chaudhry^1,2

¹Department of Biological Sciences, Oakland University, ²OU-WB Institute for Stem Cell and Regenerative Medicine, ³Department of Obstetrics and Gynecology, St. John Provindence - Providence Park Hospital, ⁴Department of Neurosurgery, Beaumont Health System

Here, we present a protocol for the dissection of human umbilical cord (UC) and fetal placenta sample into cord lining (CL), Wharton's jelly (WJ), cord-placenta junction (CPJ), and fetal placenta (FP) for the isolation and characterization of mesenchymal stromal cells (MSCs) using the explant culture technique.

Environment

An Experimental Protocol for Studying Mineral Effects on Organic Hydrothermal Transformations

Ziming Yang¹, Xuan Fu¹

¹Department of Chemistry, Oakland University

Earth-abundant minerals play important roles in the natural hydrothermal systems. Here, we describe a reliable and cost-effective method for the experimental investigation of organic-mineral interactions under hydrothermal conditions.

Neuroscience

Immunostaining of Whole-Mount Retinas with the CLARITY Tissue Clearing Method

Elizabeth J. Alessio¹, Dao-Qi Zhang¹

¹Eye Research Institute, Oakland University

Here we present a protocol to adapt the CLARITY method of the brain tissues for whole-mount retinas to improve the quality of standard immunohistochemical staining and high-resolution imaging of retinal neurons and their subcellular structures.

Biology

Isolation of Primary Mouse Retinal Pigmented Epithelium Cells

Ryan Tomaszewski^1,2, Pragya Rajpurohit³, Mei Cheng^1,2, Amany Tawfik^1,2,3

¹Eye Research Institute, Oakland University, ²Eye Research Center (OUWB)/ERC, William Beaumont School of Medicine, ³Oral Biology and Diagnostic Sciences, Dental College of Georgia, Augusta University

This manuscript describes a simplified protocol for the isolation of retinal pigmented epithelium (RPE) cells from mouse eyes in a stepwise manner. The protocol includes the enucleation and dissection of mouse eyes, followed by the isolation, seeding, and culturing of RPE cells.

Medicine

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

Khaled Elmasry^1,2,3, Mohamed Moustafa^1,2, Mohamed Al-Shabrawey^1,2

¹Eye Research Center and Department of Foundational Medical Studies, Oakland University William Beaumont School of Medicine, ²Eye Research Institute, Oakland University, ³Human Anatomy and Embryology Department, Mansoura Faculty of Medicine, Mansoura University

This protocol presents and describes steps for the isolation, dissection, culturing, and staining of retinal explants obtained from an adult mouse. This method is beneficial as an ex vivo model for studying different retinal neurovascular diseases such as diabetic retinopathy.

Methods of Studying Retinal Vessels in Health and Diseases

Mohamed Al-Shabrawey¹

¹Eye Research Center and Foundational Medical Studies Oakland University William Beaumont School of Medicine (OUWB), Eye Research Institute, Oakland University

Methods of Studying Retinal Vessels in Health and Diseases

Biology

Isolation of Primary Mouse Retinal Glial Müller Cells

Ryan Tomaszewski¹, Mohamed S. Gad¹, Paul Negoita¹, Rana Abdelkarim¹, Amany Tawfik^1,2

¹Eye Research Institute, Oakland University, ²Eye Research Center (OUWB)/ERC, William Beaumont School of Medicine

This manuscript describes a detailed protocol for isolating retinal glial Müller cells from mouse eyes. The protocol starts with enucleation and dissection of mouse eyes, followed by isolation, seeding, and culturing of Müller cells.