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Ludwig-Maximilians University of Munich

4 ARTICLES PUBLISHED IN JoVE

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.

Biochemistry

Measuring and Interpreting Oxygen Consumption Rates in Whole Fly Head Segments

Louisa Jutta Dietz¹, Anuroop Venkateswaran Venkatasubramani¹, Annika Müller-Eigner², Martin Hrabe de Angelis^3,4,5, Axel Imhof¹, Lore Becker³, Shahaf Peleg^2,6,7

¹Munich Center of Integrated Protein Science and Biomedical Center, Ludwig-Maximilians University of Munich, ²Laboratory for Metabolism and Epigenetics in Aging, Leibniz Institute for Farm Animal Biology (FBN), ³German Mouse Clinic, Helmholtz Zentrum Munich, German Research Center for Environment and Health (GmbH), ⁴German Center for Diabetes Research (DZD), ⁵Chair of Experimental Genetics, School of Life Science Weihenstephan, Technische Universität München, ⁶Laboratory for Metabolism and Epigenetics in Brain Aging, Institute of Neuroregeneration & Neurorehabilitation of Qingdao University, ⁷Molecular Biology Division, Biomedical Center, Faculty of Medicine, Ludwig-Maximilians University of Munich

Measuring alterations in metabolic rates is central to understanding the progression of various diseases and aging. Here, we present a novel technique to measure whole head oxygen consumption that more closely resembles the physiological state and may aid in revealing novel drugs that modify mitochondrial activity.

Genetics

Using Ustilago maydis as a Trojan Horse for In Situ Delivery of Maize Proteins

Isabell-Christin Fiedler¹, Arne Weiberg², Karina van der Linde¹

¹Department of Biology, Regensburg University, ²Department of Biology, Ludwig-Maximilians University of Munich

This work describes the cloning of an Ustilago maydis Trojan horse strain for the in situ delivery of secreted maize proteins into three different types of maize tissues.

Immunology and Infection

Efficient Transfection of In vitro Transcribed mRNA in Cultured Cells Using Peptide-Poloxamine Nanoparticles

Qin Xiao^*1, Yuheng Liu^*1, Dandan Zhang¹, Chao Li¹, Qihua Yang¹, Dongshui Lu¹, Weijun Zhang¹, Joseph Rosenecker², Quanming Zou¹, Yang Li³, Shan Guan¹

¹National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, Third Military Medical University, ²Department of Pediatrics, Ludwig-Maximilians University of Munich, ³Department of Pharmacy, Southwest Hospital, Third Military Medical University

A self-assembled peptide-poloxamine nanoparticle (PP-sNp) is developed using a microfluidic mixing device to encapsulate and deliver in vitro transcribed messenger RNA. The described mRNA/PP-sNp could efficiently transfect cultured cells in vitro.