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University College London Campus

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

An Improved Protocol to Purify and Directly Mono-Biotinylate Recombinant BDNF in a Tube for Cellular Trafficking Studies in Neurons

Nicolás Stuardo^1,2, Guillermo Moya-Alvarado^1,3, Carolina Ramírez^1,3, Giampietro Schiavo⁴, Francisca C. Bronfman^1,3

¹Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, ²Department of Molecular and Cellular Biology, Faculty of Biological Sciences, Center for Aging and Regeneration (CARE UC), Pontificia Universidad Católica de Chile, ³Institute of Biomedical Sciences. Faculty of Medicine and Faculty of Life Sciences, Universidad Andrés Bello, ⁴Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology and UK Dementia Research Institute at UCL, University College London Campus

Recombinant BDNF containing an Avi sequence (BDNFAvi) is produced in HEK293 cells in a cost-effective manner and is purified by affinity chromatography. BDNFavi is then directly mono-biotinylated with the enzyme BirA in a tube. BDNFavi and mono-biotinylated BDNFavi retain their biological activity when compared to commercially available BDNF.

Neuroscience

Expanding the Toolkit for In Vivo Imaging of Axonal Transport

Andrew P. Tosolini^1,2, David Villarroel-Campos^1,2,3, Giampietro Schiavo^1,2,3, James N. Sleigh^1,2,3

¹Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology, University College London, ²UCL Queen Square Motor Neuron Disease Centre, University College London, ³UK Dementia Research Institute, University College London

Using transgenic fluorescent mice, detailed protocols are described to assess in vivo axonal transport of signaling endosomes and mitochondria within motor and sensory axons of the intact sciatic nerve in live animals.