Harvard Medical School and Brigham and Women's Hospital

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Detection of MicroRNAs in Microglia by Real-time PCR in Normal CNS and During Neuroinflammation

Tatiana Veremeyko¹, Sarah-Christine Starossom¹, Howard L. Weiner¹, Eugene D. Ponomarev¹

¹Center for Neurologic Diseases, Brigham and Women's Hospital, Harvard Medical School

Microglia are resident macrophages that provide the first line of defense and immune surveillance of the central nervous system. MicroRNAs are regulatory molecules that play an important role in many physiological processes including activation and differentiation of macrophages. In this article, we describe the method for measurement of microRNAs in microglia.

Biology

DiI Perfusion as a Method for Vascular Visualization in Ambystoma mexicanum

Anna J. Saltman^*1,2, May Barakat^*1, Donald M. Bryant¹, Anastasia Brodovskaya¹, Jessica L. Whited¹

¹Department of Orthopedic Surgery, Harvard Medical School and Brigham and Women's Hospital, ²Division of Graduate Medical Sciences, Boston University

Using a lipophilic 1,1'-Dioctadecy-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) staining technique, Ambystoma mexicanum can undergo vascular perfusion to allow for easy visualization of the vasculature.

Immunology and Infection

Fecal (micro) RNA Isolation

Fyonn H. Dhang^1,2, Howard L. Weiner^1,2, Shirong Liu^1,2

¹Ann Romney Center for Neurologic Diseases, Department of Neurology, Partners Multiple Sclerosis Center, Brigham and Women's Hospital and Harvard Medical School, ²Evergrande Center for Immunologic Diseases, Harvard Medical School and Brigham and Women's Hospital

This protocol isolates high quality total RNA from fecal samples of animal and human subjects. A commercial miRNA isolation kit is used with significant adaption to isolate pure RNA with optimized quantity and quality. The RNA isolates are good for most downstream RNA assays such as sequencing, micro-array, and RT-PCR.

Neuroscience

Construction of Local Field Potential Microelectrodes for in vivo Recordings from Multiple Brain Structures Simultaneously

Anastasia Brodovskaya^*1, Shinnosuke Shiono^*1, Tamal Batabyal¹, John Williamson¹, Jaideep Kapur^1,2

¹Department of Neurology, University of Virginia, ²UVA Brain Institute, University of Virginia

The present protocol describes the construction of custom-made microelectrode arrays to record local field potentials in vivo from multiple brain structures simultaneously.