S'identifier

University of Colorado Denver School of Medicine

3 ARTICLES PUBLISHED IN JoVE

Neuroscience

Laser Capture Microdissection of Neurons from Differentiated Human Neuroprogenitor Cells in Culture

Ron Bouchard^1,2, Thomas Chong^1,2, Subbiah Pugazhenthi^1,2

¹Section of Endocrinology, Denver VA Medical Center, ²Department of Medicine, University of Colorado Denver School of Medicine

Human neuroprogenitor cells (NPCs) were expanded under proliferating conditions. NPCs were differentiated into neuron-rich cultures in the presence of a combination of neurotrophins. Neuronal markers were detected by immunofluorescence staining. To isolate a pure population of neurons, NPCs were differentiated on PEN membrane slides and laser capture microdissection was performed.

Neuroscience

Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons

Nissa L. Carrodus^*1, Kathleen Sue-Lyn Teng^*1, Kathryn M. Munro¹, Matthew J. Kennedy², Jenny M. Gunnersen^1,3

¹Department of Anatomy & Neuroscience, The University of Melbourne, ²Department of Neurobiology, Duke University Medical Center, ³Florey Institute of Neuroscience & Mental Health, The University of Melbourne

We describe a method to label protein on the surface of living neurons using a specific polyclonal antibody to extracellular epitopes. Protein bound by the antibody on the cell surface and subsequently internalized via endocytosis can be distinguished from protein remaining on, or trafficked to, the surface during the incubation.

Neuroscience

Measuring Transcellular Interactions through Protein Aggregation in a Heterologous Cell System

Susana Restrepo¹, Samantha L. Schwartz¹, Matthew J. Kennedy¹, Jason Aoto¹

¹Department of Pharmacology, University of Colorado Denver School of Medicine

Here, we present an optimized protocol to rapidly and semiquantitatively measure ligand-receptor interactions in trans in a heterologous cell system using fluorescence microscopy.