Welcome to Jon Hanley's Lab

At the University of Bristol

Molecular mechanisms of synaptic plasticity

Our Research


Learning and the formation of memories involves alterations in neural circuitry, brought about by changes in synaptic strength, which are in turn underpinned by changes in the molecular machinery of synapses. We use a range of biochemical, molecular and imaging techniques, and we collaborate with electrophysiologists for analysing synaptic function. We study three aspects of synaptic plasticity: AMPA receptor trafficking, dendritic spine morphogenesis and the control of protein translation by microRNAs.

MicroRNAs

Long-term changes in synaptic efficacy that underlie the persistent formation of memories require changes in the synthesis of synaptic proteins by the activity-dependent regulation of local translation of mRNA in dendrites close to synapses. A role for microRNAs (miRNAs) in this process has recently become the subject of much attention. MiRNAs are small, noncoding endogenous RNA molecules that repress the translation of target mRNAs through complementary binding in the transcript 3’-untranslated region (3’-UTR). A number of miRNAs have been shown to be involved in specific forms of learning and memory, and dysfunction of miRNAs is implicated in neurological and neuropsychiatric diseases including Alzheimer’s, Huntington’s and Schizophrenia.

Argonaute (Ago) proteins associate with miRNAs as well as numerous additional proteins in RNA-induced silencing complexes (RISCs) to direct translational repression of target mRNAs. We are investigating how the induction of synaptic plasticity affects this process, by studying the dynamics of RISC protein-protein interactions, how they are regulated (phosphorylation, direct binding of Ca2+, etc.) and their impact on synaptic structure and function. In a current project, we have found that the interaction between Ago2 and key RISC components including GW182 are increased by Ago2 phosphorylation in response to the induction of LTD. This mechanism leads to increased translational repression of key proteins involved in synaptic structure, and hence causes spine shrinkage (see diagram; Rajgor et al., EMBO Journal 2018).


During LTD induction, NMDAR stimulation activates signalling pathways that phosphorylate Ago2, causing increased binding to GW182 and a consequent increase in translational repression. This reduces the expression of proteins involved in regulating the actin cytoskeleton, causing a decrease in spine size. ORF=open reading frame; rib=ribosome.


We have also demonstrated that PICK1 inhibits miRNA activity by binding to Ago2 on endosomal compartments in dendrites (Antoniou et al., 2014, EMBO Reports). This interaction is disrupted by Ca2+ions, and therefore NMDAR-dependent LTD causes a dissociation of Ago2 from PICK1, increasing miRNA activity and hence translational repression (Rajgor et al., 2017, J.Biol.Chem). 

We are also very interested in how these mechanisms of RISC regulation are affected by pathologies such as Alzheimer's and ischaemia. 

We have recently (March 2019) secured an Alzheimer's Research UK (ARUK) grant to study the role of RISC regulation in dendritic spine loss and synaptic dysfunction in mouse models of AD. 

  

  

  

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AMPAR trafficking

AMPA receptors (AMPARs) mediate the majority of fast excitatory synaptic transmission in the brain, and plasticity at excitatory synapses involves alterations in the number of AMPARs localised at the synaptic plasma membrane, brought about by regulated receptor trafficking. AMPAR expression at the synaptic plasma membrane is regulated by endocytosis, exocytosis, recycling and lateral diffusion events that contribute to reductions (Long Term Depression, LTD) or increases (Long Term Potentiation, LTP) in synaptic strength.

A major focus of the lab is investigating how basic cell biological processes interact with AMPAR subunits to bring about changes in AMPAR trafficking and hence synaptic strength. In particular, we are studying a PDZ- and BAR-domain protein called PICK1, which binds GluA2/3 subunits of AMPARs, and is involved in AMPAR internalisation and LTD. We have demonstrated that PICK1 inhibits Arp2/3-mediated actin polymerisation, and that this is required for NMDA-stimulated AMPAR internalisation and LTD (Rocca et al., 2008, Nat. Cell Biol; Nakamura et al., 2011, EMBO J.; Rocca et al., 2013, Neuron). We are also studying how PICK1 interacts with core components of the endocytic machinery to regulate clathrin-coated vesicle formation (Fiuza et al., 2017, J. Cell Biol.). 


Increase in endogenous AMPAR internalisation in cultured hippocampal neurons in response to stimulation with NMDA.



Schematic representation of AMPAR trafficking pathways involved in LTD or LTP.



As well as “normal” plasticity, we are studying AMPAR trafficking events that occur in response to acute injury, such as mechanical injury (a model for traumatic brain injury, TBI) and oxygen/glucose deprivation (OGD; a model for ischaemia). Brain ischaemia occurs when the blood supply to the brain is interrupted, for example by occlusion following a stroke, or as a result of cardiac arrest. We are studying the specific endosomal sorting steps that are involved in regulating cell-surface AMPARs in response to OGD or TBI. The majority of AMPARs contain GluA2 subunit, and consequently are Ca2+ impermeable. OGD and TBI cause trafficking events that result in the loss of surface GluA2-containing AMPARs, and therefore an increase in Ca2+ permeable AMPARs, which contribute to neuronal death. We have discovered differences in subunit-specific AMPAR trafficking events between hippocampal and cortical neurons that we propose play a role in the differential vulnerabilities to ischaemia between these neuronal types observed in vivo (Blanco-Suarez et al., 2014, J. Biol. Chem; Koszegi et al., 2017, Scientific Reports).

  

  

  

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Dendritic spine morphogenesis

Dendritic spines are highly specialized subcellular compartments that contain the postsynaptic protein machinery of most excitatory synapses. They concentrate and compartmentalise biochemical signals such as Ca2+, and synaptic protein machinery such as neurotransmitter receptors, providing the synaptic specificity required for plasticity. Changes in synaptic strength correlate with corresponding changes in dendritic spine size, and possibly shape. LTD stimuli result in spine shrinkage and retraction, whereas LTP leads to the formation of new spines, or the growth of existing ones.

 

 

A dendrite from a hippocampal neuron expressing GFP-actin, which is enriched in spine heads.


 
We are studying the molecular mechanisms that underlie changes in spine size during synaptic plasticity (Nakamura et al., 2011, EMBO J.; Rocca et al., 2013, Neuron) and also in response to OGD (Blanco Suarez et al., 2014, J.Cereb. Blood Flow Metab.).

Spines are extremely rich in dynamic actin filaments, which underlie these activity-dependent changes in spine size. LTP is thought to promote actin polymerization resulting in an increase in spine F-actin, and LTD results in reduced F-actin via actin depolymerization. The signalling pathways that lead to these changes in actin polymerization to bring about spine growth or shrinkage are affected by OGD, and we propose that manipulating these pathways holds therapeutic potential.

 

 

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People


Omar Kassaar

Post-doc

MicroRNAs and Argonaute phosphorylation

Georgiana Stan

4th year PhD student

AMPAR trafficking, PICK1, FLIM-FRET imaging

Jess Walters

2nd year PhD student

Argonaute and miRNAs in Alzheimer's models

Fathima Murshida

1st year PhD Student

Argonaute, miRNA

Agnieszka Moskal

MRes student

DDX6, miRNA

Suko Nakamura

Super-tech/ lab manager

Jon Hanley

PI

Publications


Key publications from our lab:

2018

Rajgor D., Sanderson T.M., Amici M., Collingridge G.L., and Hanley J.G. (2018) NMDAR-dependent Argonaute 2 phosphorylation regulates miRNA activity and dendritic spine plasticity. The EMBO Journal. https://www.ncbi.nlm.nih.gov/pubmed/29712715

Parkinson G.T., Chamberlain S.E.L.,  Jaafari N., Turvey M., Mellor J.R. and  Hanley J.G. (2018) Cortactin regulates endo-lysosomal sorting of AMPARs via direct interaction with GluA2 subunit. Scientific Reports. https://www.ncbi.nlm.nih.gov/pubmed/29515177


2017

Smith K.R., Rajgor D., and Hanley J.G. (2017). Differential regulation of the Rac1 GTPase activating protein (GAP) BCR during oxygen/glucose deprivation in hippocampal and cortical neurons. The Journal of Biological Chemistry.  https://www.ncbi.nlm.nih.gov/pubmed/29046349

Koszegi Z., Fiuza M., and Hanley J.G. (2017). Endocytosis and lysosomal degradation of GluA2/3 AMPARs in response to oxygen/glucose deprivation in hippocampal but not cortical neurons. Scientific Reports. https://www.ncbi.nlm.nih.gov/pubmed/28951554

Fiuza M., Rostosky C.M., Parkinson G.T., Bygrave A.M., Halemani N., Baptista M., Milosevic I., and Hanley J.G. (2017). PICK1 regulates AMPA receptor endocytosis via direct interactions with AP2 α-appendage and dynamin. Journal of Cell Biology. https://www.ncbi.nlm.nih.gov/pubmed/28855251

Rajgor D, Fiuza M, Parkinson GT, Hanley JG. (2017). PICK1 Ca2+ Sensor Modulates NMDA Receptor-Dependent MicroRNA-Mediated Translational Repression in Neurons. The Journal of Biological Chemistry 292(23):9774-9786. https://www.ncbi.nlm.nih.gov/pubmed/28404816


2015

Cockbill LM, Murk K, Love S, Hanley JG (2015) Protein interacting with C kinase 1 suppresses invasion and anchorage-independent growth of astrocytic tumor cells. Molecular Biology of the Cell 26:4552-6. https://www.ncbi.nlm.nih.gov/pubmed/26466675


2014

Blanco-Suárez E, Fiuza M, Liu X, Chakkarapani E, Hanley J.G. (2014). Differential Tiam1/Rac1 activation in hippocampal and cortical neurons mediates differential spine shrinkage in response to oxygen/glucose deprivation. The Journal of Cerebral Blood Flow and Metabolism 34:1898-906. https://www.ncbi.nlm.nih.gov/pubmed/25248834

Antoniou A., Baptista M., Carney N.C. and Hanley J.G. (2014). PICK1 links Argonaute 2 to endosomes in neuronal dendrites and regulates miRNA activity. EMBO Reports 15:548-56. https://www.ncbi.nlm.nih.gov/pubmed/24723684
         
Blanco-Suárez E. and Hanley J.G. (2014). Distinct Subunit-specific α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) Receptor Trafficking Mechanisms in Cultured Cortical and Hippocampal Neurons in Response to Oxygen and Glucose Deprivation. The Journal of Biological Chemistry 289:4644-51. https://www.ncbi.nlm.nih.gov/pubmed/24403083

Hanley J.G. (2014). Actin-dependent mechanisms in AMPA receptor trafficking. Frontiers in Cellular Neuroscience, 8:381. REVIEW. https://www.ncbi.nlm.nih.gov/pubmed/25429259

Hanley JG. (2014). Subunit-specific trafficking mechanisms regulating the synaptic expression of Ca2+-permeable AMPA receptors. Seminars in Cell and Developmental Biology, 27:14-22. REVIEW.  https://www.ncbi.nlm.nih.gov/pubmed/24342448

2013

Murk K., Blanco Suarez E.M., Cockbill L.M.R., Banks P., and Hanley J.G. (2013). The antagonistic modulation of Arp2/3 activity by N-WASP/WAVE2 and PICK1 defines dynamic changes in astrocyte morphology. Journal of Cell Science 126:3873-83. https://www.ncbi.nlm.nih.gov/pubmed/23843614

Rocca D.L., Amici A., Antoniou A., Blanco Suarez E., Halemani N., Murk K., McGarvey J., Jaafari N., Mellor J.R., Collingridge G.L., and Hanley J.G. (2013).  The small GTPase Arf1 regulates Arp2/3-mediated actin polymerization via PICK1 to control synaptic plasticity.  Neuron 79:293-307. https://www.ncbi.nlm.nih.gov/pubmed/23889934


2012

Jaafari, N., Henley, J.M., and Hanley, J.G. (2012). PICK1 mediates transient synaptic expression of GluA2-lacking AMPARs during glycine-induced AMPA receptor trafficking. Journal of Neuroscience 34, 11618-30. https://www.ncbi.nlm.nih.gov/pubmed/22915106


2011

Dennis S.H., Jaafari N., Cimarosti H., Hanley J.G., Henley J.M., and Mellor J.R. (2011). Oxygen/Glucose Deprivation Induces a Reduction in Synaptic AMPA Receptors on Hippocampal CA3 Neurons Mediated by Metabotropic Glutamate Receptor 1 and A3 Receptors. Journal of Neuroscience 31, 11941-52. https://www.ncbi.nlm.nih.gov/pubmed/21849555
           
Nakamura Y., Wood C.L., Patton A.P., Jaafari N., Henley J.M., Mellor J.R., and Hanley J.G. (2011). PICK1 inhibition of the Arp2/3 complex controls dendritic spine size and synaptic plasticity. The EMBO Journal, 30:719-30. https://www.ncbi.nlm.nih.gov/pubmed/21252856


2010

Hanley JG. (2010). Endosomal sorting of AMPA receptors in hippocampal neurons. Biochemical Society Transactions, 38: 460-465. REVIEW. https://www.ncbi.nlm.nih.gov/pubmed/20298203


2009

Dixon R.M., Mellor J.R. and Hanley J.G. (2009). PICK1-Mediated Glutamate Receptor Subunit 2 (GluR2) Trafficking Contributes To Cell Death In Oxygen/Glucose Deprived Hippocampal Neurons. The Journal of Biological Chemistry, 284: 14230-5. https://www.ncbi.nlm.nih.gov/pubmed/19321442


2008

Rocca D.L., Martin S., Jenkins E.L. and Hanley J.G. (2008). Inhibition of Arp2/3-mediated actin polymerisation by PICK1 regulates neuronal morphology and AMPA receptor endocytosis. Nature Cell Biology, 10: 259-271. https://www.ncbi.nlm.nih.gov/pubmed/18297063

Hanley JG. (2008). AMPA receptor trafficking pathways and links to dendritic spine morphogenesis. Cell Adhesion and Migration, 2: 276-82. REVIEW. https://www.ncbi.nlm.nih.gov/pubmed/19262155

Hanley J.G. (2008). PICK1: a multi-talented modulator of AMPA receptor trafficking. Pharmacology and Therapeutics, 118: 152-60. REVIEW. https://www.ncbi.nlm.nih.gov/pubmed/18353440

2007

Hanley J.G. (2007). NSF Binds Calcium to Regulate its Interaction with AMPA Receptor Subunit GluR2. Journal of Neurochemistry, 101: 1644-50. https://www.ncbi.nlm.nih.gov/pubmed/17302911


2005

Hanley J.G., and Henley J.M. (2005).  PICK1 is a calcium-sensor for NMDA-induced AMPA receptor trafficking. The EMBO Journal, 24: 3266-78. https://www.ncbi.nlm.nih.gov/pubmed/16138078


2002

Hanley J.G., Khatri L., Hanson P.I., and Ziff E.B (2002).  NSF ATPase and a/b SNAPs disassemble the AMPA receptor-PICK1 complex. Neuron, 34: 53-67. https://www.ncbi.nlm.nih.gov/pubmed/11931741


2000

Billups D*., Hanley J.G*., Orme M., Attwell D., and Moss S.J. (2000). GABAC receptor sensitivity is modulated by interaction with MAP1B. Journal of Neuroscience, 20: 8643-8650. https://www.ncbi.nlm.nih.gov/pubmed/11102469
* 1st equal authorship

Hanley J.G., Jones E.M., and Moss S.J. (2000). GABA receptor rho1 subunit interacts with a novel splice variant of the glycine transporter, GLYT-1. Journal of Biological Chemistry, 275: 840-846. https://www.ncbi.nlm.nih.gov/pubmed/10625616


1999

Hanley J.G., Koulen P., Bedford F., Gordon-Weeks P.R., and Moss S.J. (1999). The protein MAP-1B links GABA(C) receptors to the cytoskeleton at retinal synapses. Nature, 397: 66-69. https://www.ncbi.nlm.nih.gov/pubmed/9892354

Join the Lab


PhD studentship available!

A 4 year BBSRC-funded PhD studentship offered under the SWBio DTP to study the local control of synaptic protein synthesis in neurons by microRNA.

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  • School of Biochemistry, University Walk, Bristol, BS8 1TD
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