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The Molecular NeuroPathobiology Laboratory
The research programme of the Molecular NeuroPathobiology Laboratory (MNP) is focussed on proving the hypothesis that impairment of the selectivity and/or efficiency of long-range communication in neurons caused by defects in membrane traffic constitute a major pathogenic mechanism in the nervous system. Indeed, the unique architecture and signalling capacity of neurons is maintained by trafficking events regulating the dynamics of intracellular organelles, such as signalling endosomes and mitochondria, and the spatio-temporal pattern of receptor stimulation by morphogens, neurotrophins and other growth factors. Axonal transport constitutes the backbone of this long-distance crosstalk, functionally connecting peripheral compartments, such as synapses, with events occurring in the soma and vice versa.
Our laboratory has provided crucial contributions to the field by defining the mechanism(s) responsible for the uptake and sorting of neurotrophin receptor complexes and virulence factors, and the coupling of these endocytic routes with the axonal retrograde transport pathway. We have demonstrated the essential role of cytoplasmic dynein and its adaptor proteins in this process, and provided a functional link between mutations in components of this transport pathway and neuronal dysfunctions. The translational potential of our findings have recently been confirmed by the discovery that these mutations are associated with a broad spectrum of disorders of the nervous system in human patients, including motor and sensory pathologies.
Figure 1. Primary spinal cord motor neuron in culture
Cellular Models: Our main model is the motor neuron, a type of neuron that functionally connect skeletal muscles with motor control centres located in the CNS. We isolate motor neurons either from embryonic spinal cord or derive them from mouse embryonic stem (ES) cells by in vitro differentiation. In addition, we also culture dorsal root ganglia, cortical, and hippocampal neurons.
Approaches: We have set up in our laboratory several techniques enabling us to study the composition, trafficking and signalling potential of signalling endosomes. We have performed imaging-based chemical and siRNA screens to test modulators of axonal transport and somatic sorting using ES-derived motor neurons, and established imaging protocols to monitor axonal transport both in vitro and in vivo using high resolution confocal and electron microscopy.
We strongly believe that only by understanding the basic principles of membrane trafficking and signalling will we be able to uncover the pathological changes responsible for many human disorders, including cancer. These fundamental processes might then be exploited to provide an earlier and more accurate diagnosis, translating into improved pharmacological treatments and more favourable prognosis.
Professor Giampietro Schiavo's UCL IRIS Profile
Current Projects
MNP Team Members
Image Archive
Figure 2 and 3. Differentiation of mouse ES cells into MNs. An optimised protocol allows the differentiation of primary MNs from a mouse ES cell line. MN specification is determined by the addition of retinoic acid and sonic hedgehog, whilst MN maturation is achieved in the presence of neurotrophins. ES cell-derived MNs are characterized by an HB9-driven GFP staining (green) and are positive for the bIII isoform of tubulin (red), and for the dendritic marker MAP2 (blue).
Figure 2 and 3. Differentiation of mouse ES cells into MNs. An optimised protocol allows the differentiation of primary MNs from a mouse ES cell line. MN specification is determined by the addition of retinoic acid and sonic hedgehog, whilst MN maturation is achieved in the presence of neurotrophins. ES cell-derived MNs are characterized by an HB9-driven GFP staining (green) and are positive for the bIII isoform of tubulin (red), and for the dendritic marker MAP2 (blue).
Figure 4 and 5. Mouse neuromuscular junctions (NMJ). In Figure 4, a post-synaptic marker (alpha-bungarotoxin - BTx; in red) highlight a group of NMJs. Figure 5 shows a close up of a NMJ and its axonal shaft stained with an axonal retrograde transport probe (in red) and BTx (in green).
Figure 1. Spinal cord motor neurons control muscle contraction by electrical output signals. This figure shows the distribution of TeNT HC (blue), the Neurotrophin receptor p75NTR (red) and BDNF (green) in differentiated MN in culture.
Figure 6. 3D-reconstruction by microtomography of the heart of a Kidins220/ARMS-/- embryo. Note the dilated atria and the altered outflow tract. Images collected and analysed by I.R. Orriss (UCL Institute of Ophthalmology), A. Weston and L. Collinson (Electron Microscopy Facility, LRI).
Figure 8. Rab7 and peripherin are co-expressed in sensory neurons. Peripherin displays a filamentous distribution in the axonal network of embryonic dorsal root ganglia (DRGs) in culture (in green), whereas Rab7 shows a very intense punctate staining in the soma and in the network. At close inspection, Rab7-positive organelles are often in close apposition to peripherin filaments. Scale bars, 10 µm.
Figure 9. High-resolution mapping of axonal signalling endosomes. Confocal image of embryonic stem cells-derived motor neurons incubated with HCT conjugated to monocrystalline iron oxide nanoparticles (MIONs) for 1 h at 37˚C. HB9 motor neuron-specific promoter is depicted in green, the HA tag of TeNT HCT in red and bIII-tubulin in blue. Scale bar: 5 μm.
Figure 4 and 5. Mouse neuromuscular junctions (NMJ). In Figure 4, a post-synaptic marker (alpha-bungarotoxin - BTx; in red) highlight a group of NMJs. Figure 5 shows a close up of a NMJ and its axonal shaft stained with an axonal retrograde transport probe (in red) and BTx (in green).
Figure 7. Kidins220/ARMS is required for capillary plexus development in the brain. A section of E16.5 Kidins220/ARMS-/- brain was stained with an endothelial cell marker (in green) and for active caspase-3 (in red).
- Grant support
The MNP laboratory was supported three Cancer Research UK (CRUK) core grants, which supported my research programme from the creation of my laboratory at the CRUK London Research Institute (LRI) in 1997.
2016 The Royal Society Newton International Fellowship. Neurotrophins and the balance between retrograde signalling and autophagic flux in the axon. To O Marcelo Lazo.
2016 MRC Programme Grant. Novel and bespoke mouse models for dissecting neurodegenerative disease. MR/N501931/1. To: EL Fisher, L Greensmith, G Schiavo, A Isaacs, P Fratta, F Wiseman and A Acevedo. £1.78M for four years
2016 Horserace Betting Levy Board. Why do horses roar? From the beginning to the end of recurrent laryngeal neuropathy. To: R Piercy, A. Draper, J. Cheetham, G Schiavo, S Brown and J Perkins. £97,867 for 24 months
2016 NC3Rs Fellowship. A new Drosophila-based strategy to study mitochondrial transport and neuronal ageing in vivo. To: Dr A Vagnoni. £195,000 for 36 months
2016 The Wellcome Trust Postdoctoral Training Fellowship for Clinicians “Control of receptor trafficking as a therapeutic target in the inherited neuropathies” 110043/Z/15/Z. To: A Rossor and G Schiavo. £412,604 for 36 months
2016 MNDA PhD Fellowship. Regulation of intracellular traffic by TBK1 and its relevance to Amyotrophic Lateral Sclerosis. To: G Schiavo and P Fratta. £92,842 for three years
2015 MRC Clinical Fellowship. Investigating deficits of axonal RNA metabolism and axonal signalling in ALS. To P. Fratta. £1,156,170 for four years
2015 Fondation Thierry Latran. Deficits in axonal transport as a target for pharmacological intervention in Amyotrophic Lateral Sclerosis. To: G Schiavo and L Greensmith. €117,000 for 18 months
2015 The Wellcome Trust Senior Investigator Award. The mechanism controlling sorting and axonal retrograde transport in neurons. £1.75M for five years
2014 Alzheimer’s Research UK – UCL Drug Discovery Institute. Lead Academic Scientist (with B. De Strooper and J. Hardy). £10M for 5 years
2014 Medical Research Council’s UK Dementias Platform (UKDP). Co-PI (UCL Chairman: T.T. Warner). £1.9M for the set-up of a new high-throughput phenotypic cell screening facility
2014 BBSRC Industrial CASE PhD Fellowship. Mechanisms controlling the axonal retrograde transport pathway. To: G Schiavo and L Greensmith. 48 months.
2013 UCL SLMS and ION Capital Equipment Fund for the purchase of a multiphoton confocal microscope for intravital imaging. £437.000
2013 Assembly of neuromuscular circuits from stem-cell derived components in vitro (R130577). I. Lieberman, G Schiavo and L Greensmith.
2013 7th Framework Programme European Commission. FP7-PEOPLE-2009-IEF. Proposal n° 329826. Assembling a functional map of axonal signalling endosomes. €212.000 for 24 months.
2012 Cancer Research UK Core grant. Understanding axonal transport and somatic sorting in health and disease. £1.50M
2011 MRC Programme grant (co-applicant with S Tabrizi). Cellular pathophysiology of prion-mediated neurodegeneration – a model for understanding protein misfolding disorders. £603,000 of which £7,175 to GS.
2011 Sir Henry Wellcome Fellowship (to Dr A Twelvetrees). The Welcome Trust. The coordination of MT-dependent motors during axonal transport. £250,000.00 for 48 months
2011 BBSRC Industrial CASE PhD Fellowship. Modifying axonal transport as a therapeutic strategy for ALS. To: G Schiavo and L Greensmith. £84,000 for 48 months.
2011 Weizmann UK Joint Research Program. Motor-Driven Transcription Factors in Injured Nerve – How Fast Can They Go? To: M. Fainzilber and G Schiavo. £29,994.41 for 24 months to GS.
2011 Tokyo University of Science. Travel grant to visit the laboratory of Prof T Nakamura. ¥300,000.
2011 7th Framework Programme European Commission. FP7-PEOPLE-2009-IEF. Proposal n° 252860. The role of Rab7 in axonal retrograde transport and human pathologies. £146,786.93 for 24 months.
2010 MRC Industrial CASE PhD Fellowship. Neurotrophic receptor p75 and its role in neuropathic pain. To: G Schiavo and S Westbrook. £92,317 for 48 months.
Collaborators
- Professor Fabio Benfenati, Italian Institute of Technology and University of Genova, Genova (I)
- Professor Matteo Caleo, Universita di Padova (I)
- Professor Catarina Brito, iBET, Oeiras (P)
- Professor Cecilia Bucci, University of Salento, Lecce (I)
- Dr Fabrizia Cesca, Italian Institute of Technology and University of Genova, Genova (I)
- Professor Mike Fainzilber, Weizmann Institute of Science, Rehovot (Is)
- Professor Elizabeth Fisher, UCL – Institute of Neurology, London (UK)
- Professor Linda Greensmith, UCL – Institute of Neurology, London (UK)
- Professor Eric Kremer, Institut de Génétique Moléculaire de Montpellier, Montpellier (F)
- Professor Nicholas Mazarakis, Imperial College London (UK)
- Professor Cesare Montecucco, University of Padova, Padova (I)
- Dr Laura Restani, CNR Neuroscienze di Padova (I)
- Dr Andrea Serio, King's College London (UK)