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University of Wuerzburg

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Lectin-based Isolation and Culture of Mouse Embryonic Motoneurons

Rebecca Conrad¹, Sibylle Jablonka², Teresa Sczepan¹, Michael Sendtner², Stefan Wiese¹, Alice Klausmeyer¹

¹Institute for Cellmorphology and molecular Neurobiology, Group for Cellbiology, Ruhr-University Bochum, ²Institute for Clinical Neurobiology, University of Wuerzburg

An alternative way of isolating mouse embryonic motoneurons from the spinal cord is described. The method takes into account the fact that lectin can bind to the low affinity nerve growth factor receptor p75NTR. This lectin-based preplating allows a purification similar to that with a specific antibody against the p75NTR.

Biology

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)

Samira Samtleben¹, Juliane Jaepel^1,2, Caroline Fecher¹, Thomas Andreska¹, Markus Rehberg³, Robert Blum¹

¹Institute for Clinical Neurobiology, University of Wuerzburg, ²Department of Synapses - Circuits - Plasticity, Max Planck Institute of Neurobiology, Martinsried, ³Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians University of Munich

Targeted-esterase induced dye loading (TED) supports the analysis of intracellular calcium store dynamics by fluorescence imaging. The method bases on targeting of a recombinant Carboxylesterase to the endoplasmic reticulum (ER), where it improves the local unmasking of synthetic low-affinity Ca²⁺ indicator dyes in the ER lumen.