https://www.twelvetreeslab.co.uk/home daily 1.0 2024-10-02 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507563282628-OFEDGWI1ECL10ERQIVDG/buttonArtboard+3-100.jpg Home - Approaches https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507985628242-4CH68G59VWKLUSVL89M5/buttonArtboard+7%402x-100.jpg Home https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508418841580-NXYQHBT55IC8FE56VBKH/K560.gif Home Individual kinesin molecules walking along immobilised microtubules in vitro. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508417875656-A60BC0BJJO7578Q6SGOW/dynein-GFP+overnight.gif Home Dynein-GFP oscillating between developing neurites imaged over 14 hours. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508417823407-SZ7HZ5W5RHD29UVK7AEI/EB3_dendrites_orange.gif Home EB3 comets falling in a dendritic tree. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507985594511-XYO4O0MJKMW80EJETA4F/buttonArtboard+5%402x-100.jpg Home https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508418930957-SLQYCRYY9IMVQ0PV6M8B/DIC1+TAU+8DIV.jpg Home Dynein-GFP (cyan) accumulating in the distal axons labelled with for tau (orange). https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507563300440-BGVAZ0DYVRX83I28HM7F/buttonArtboard+4-100.jpg Home - Papers https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507563249329-HA0OD8N3AGZHIJ7Z2BD7/buttonArtboard+1-100.jpg Home https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507129618814-H2F8T7T89U6S9GQ2W6XG/plate.jpg Home https://www.twelvetreeslab.co.uk/papers daily 0.75 2021-03-18 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507989681867-B0PWX1NAYT16XBI57ZDV/miroArtboard+1%402x-100.jpg Papers https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507039041602-GTMD20RABXE7K6DE21KT/DynDynArtboard+1%402x-100.jpg Papers - Dynein accumulates in the distal axon Our first discovery when we cultured hippocampal neurons from the dynein-GFP mouse, was pronounced accumulation of dynein in the the distal axon. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507039040404-VZR8NYW2KGDGD6W3A1VY/DynDynArtboard+2%402x-100.jpg Papers - Dynein accumulates in the growth cone coincident with axon specification When we imaged neuronal development over many hours, dynein accumulation was dynamic between neurites. However, dynein was present coincident with axon speification. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507039042169-5Q8DGBXGNOPP054LQPTT/DynDynArtboard+3%402x-100.jpg Papers - Individual anterograde transport events of dynein are visible by HiLo imaging. Kymographs of axons imaged by near TIRF (HiLo) show dynein moving by short anterograde events of comparatively uniform velocity and low signal intensity. Typical speed was ~1 um/s, equivalent to the known speed of kinesin-1. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507039043737-TP5MBRQART0M6OE9CBA7/DynDynArtboard+4%402x-100.jpg Papers - Neuron specific isoforms of dynein interact directly with the kinesin light chain Using extensive biochemistry we found that the direct interaction between dynein and kinesin was mediated by neuron specific isoforms of the dynein intermediate chain. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507039043845-X1KK3BYEV9AVL0327IBO/DynDynArtboard+5%402x-100.jpg Papers - Anterograde transport of dynein is dependent on the direct interaction with kinesin-1 Neurons treated with a peptide blocking the interaction between kinesin and dynein showed a large reduction in dynein anterograde trasnport speeds. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507039044768-41MEOK6AXYF07VLHFYDY/DynDynArtboard+6%402x-100.jpg Papers - The "kinesin-limited" model of slow axonal transport We describe a model of slow axonal transport whereby kinesin is essentially in rate limiting supply for slow transport cargoes, producing short bursts of motility and low cargo net transport speeds. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507986600782-9JAAKS3M883D10D9BZ9D/snapshotArtboard+1%402x-100.jpg Papers https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507989526912-0QZNKQ1HER7ENKHC45WW/gabaArtboard+1%402x-100.jpg Papers https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507986086716-2PA9OGB9LL53D9APS3T5/ATreviewArtboard+1%402x-100.jpg Papers https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507986419456-67IO2IU2JO4B2SW63LIA/DynMouseArtboard+1%402x-100.jpg Papers https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1506952970364-S8KZMZRQJVZR72NS3I4A/kindynArtboard+3%402x-100.jpg Papers https://www.twelvetreeslab.co.uk/about-1 daily 0.75 2022-09-30 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331277689-WLTNCS7DTPFC7IBXU0HY/IMG_4905.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/3248b59d-b911-41d5-9dc9-47d5f3ae123b/IMG_9678.jpg About - Make it stand out Whatever it is, the way you tell your story online can make all the difference. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/6bf76649-3ca2-43f2-bf2a-baae97c50145/IMG_6530.jpg About - Make it stand out Whatever it is, the way you tell your story online can make all the difference. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331332203-TJGEB9MY2DPUCOHE44GA/IMG_4951.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331333299-XHI9ZFG1RIXFHR72EXM9/IMG_4966.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/58237559-9e6a-422b-917c-28d77c6974c1/IMG_0416.jpg About - Make it stand out Whatever it is, the way you tell your story online can make all the difference. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331400849-O4RT7T12QN7E8CLHODZH/IMG_2911.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331412946-4ORTYKI3IKS8AR2YAU03/IMG_4335.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331403022-B8RN61CQL2OB229K14G3/IMG_2918.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331410830-NWSP2O8VVZ0PZY6ROVSO/IMG_3993.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331396100-GQ4PN62WB3WJK2A8KRTI/IMG_2820.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331395885-OM7CFLQRTX3XQ1SCU5DG/IMG_2823.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331405798-LACGGG3C3PKN1XXFMK5E/IMG_3974.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331411728-DJQ280YVZ67DO5AO9OBG/IMG_4258.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331418231-R1D9DZ3TJ0DSMRRU6D64/IMG_4554.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508331413837-K6TUS4DQQRAJP2QE7IDR/IMG_4560.JPG About https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507990762308-JUENYAXNO0DMY1T0XUR4/epp_tip_twoArtboard+1%402x-100.jpg About https://www.twelvetreeslab.co.uk/approaches daily 0.75 2018-10-15 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507805744118-LWUJM3A23MG4WARSG4HB/DIC1+TAU+8DIV.jpg Approaches - Mouse hippocampal neurons, 8 DIV Primary cultured neurons from the dynein-GFP mouse showing cytoplasmic dynein (cyan) and the immunolabelled axonal marker Tau (orange). https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507802931745-IY8BOGCFLYWCS95OV1QP/PSD95+MAP2+21DIV+crop.jpg Approaches - Mouse hippocampal neurons, 21 DIV Primary cultured neurons immunolabelled with MAP2 (magenta) and the excitatory post-synaptic marker PSD-95 (cyan). https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507806753835-K5B90M4YJBWNACTPQWL8/DIC1+Synapsin+8DIV.jpg Approaches - Mouse hippocampal neurons, 8 DIV Primary cultured neurons from the dynein-GFP mouse showing cytoplasmic dynein (cyan) and the immunolabelled pre-synaptic maker synapsin (orange). https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508409616589-8718D4NYTNTU6X7ARPQO/microfArtboard+1%402x-100.jpg Approaches - Microfluidic chamber Schematic diagram of a microfluidic chamber. Neurons are plated into the somal side. The microgrooves are too narrow for neuronal cell bodies to enter and so as the culture grows, only axons cross the grooves to reach the distal axonal side. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508407183255-I63W10J400K83ATJAU41/IMG_2247.png Approaches - Neurons on the somal side of the chamber A phase contrast image of primary hippocampal neurons a few days after plating in a microfluidic chamber. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508407192107-D3H3H4TR3OW1G7CB92TW/microfluidic.png Approaches - Neurons near the microgrooves A phase contrast image of primary hippocampal neurons a few days after plating in a microfluidic chamber at a higher magnification. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508407270336-OBT4OPG9VVFTF66ZLIPR/axons.png Approaches - Axons on the other side A phase contrast image of the axons of primary hippocampal neurons having crossed the microfluidic grooves a few days after plating. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507814210526-4MRSN6IC2BHAK3OT4HRJ/K560+cropped+area.gif Approaches - Kinesin motility in vitro Individual kinesin motors (cyan) walking along immobilised microtubules (magenta) imaged by single molecule TIRF microscopy. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508333595109-L8WFXGMIRC3QQMY8OENY/tirfArtboard+1%402x-100.jpg Approaches - TIRF imaging Schematic diagram of the experimental set up for TIRF imaging of microtubule motors in vitro. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508334101276-1KUXRDAEEESW453JPKTG/K560+tracking.gif Approaches - Tracking molecules Spot detection and frame to frame tracking of the motility of single kinesin molecules moving along a microtubule. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507813740871-TXI2T7NR3SX0PGASTAYF/EB3_dendrites.gif Approaches - EB3 comets Bi-directional EB3-GFP comets moving through the dendrites of a cultured neuron, captured over a few minutes. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507813737386-NXSEA94QIRNTNFPW4FHX/outgrowth.gif Approaches - Neurite development Neurite outgrowth captured over several hours. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507813732118-HTZTMQK9NH47B1PGB8F5/endosome_square.gif Approaches - Fast axonal transport Tracked vesicles moving in an isolated axon. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507570002758-K0MVQLZDNK0MYUNURZ8L/buttonArtboard+1%402x-100.jpg Approaches https://www.twelvetreeslab.co.uk/projects daily 0.75 2017-10-18 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508329469689-MPW5NHW2GM0GINY4OJ6H/dyn-dynArtboard+17%402x-100.jpg Projects Slow transport of cytoplasmic dynein populations A summary of the novel analysis strategy designed to study the slow transport of cytoplasmic dynein. This study was carried out on primary cultured neurons from the dynein-GFP mouse. Axons were isolated in microfluidic devices and the movement of discrete populations of dynein-GFP were tracked through time. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508323654485-Y1QNR3L7ITSNRRIVMXFO/kinesinArtboard+1%402x-100.jpg Projects A cartoon depicting the heavy chain of kinesin-1 unfolding. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508324037624-SW6XUUKSKX3LHDPFGEBH/DynDynArtboard+6%402x-100.jpg Projects A limited supply of kinesin motors means that slow cargos are only transported short distances before they are released, whilst the kinesin continues it's processive motility with a new cargo molecule. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508319463729-GUEUUJWNMTHJS43VCV1N/DynDynArtboard+4%402x-100.jpg Projects Isoforms of the cytoplasmic dynein intermediate chain (DIC). There are two genes for DIC, dync1i1 and dync1i2 (producing proteins DIC1 and DIC2 respectively), which are both alternatively spliced. All cells express the blue isoform, DIC2c. However the DIC1 gene is specific to mammalian neurons, as are all other splice-forms of DIC2. Click to enlarge. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1507566899691-ANY9WKX4XFWM1LQP0L61/neuronArtboard+15%402x-100.jpg Projects https://www.twelvetreeslab.co.uk/the-team daily 0.75 2024-10-14 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1632306217006-UQH3D2XQP5C5N0COQUS6/Twelvetrees-lab-Dec-2019 The Team - Make it stand out A Christmas lab selfie from December 2019 https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1586169957650-81M5JAFMGDAJ9RI5DSXC/Emma.jpg The Team https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/c6cbc271-0ab8-447b-af3f-feaf1c28a1ce/img_3038.jpg The Team - Make it stand out Whatever it is, the way you tell your story online can make all the difference. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1539638012334-RS5O35G9FNNLOE5UX9YR/Ashleigh.JPG The Team https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1539638045846-46TMDY3A3B129INEPDYK/Evie.png The Team https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508147189817-SJ0DTPAGWG6QSXEML8D5/questionArtboard+8%402x-100.jpg The Team https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/99e363ef-db6d-433f-998b-ea106b59adab/photo+for+bio.jpeg The Team - Make it stand out Whatever it is, the way you tell your story online can make all the difference. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/a6a5c724-73e1-49f3-b04a-6dfa1938b335/IMG_5641.jpg The Team - Make it stand out Whatever it is, the way you tell your story online can make all the difference. https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1539638111739-PL6FMGQIAUGB1O43RPYR/12trees.jpg The Team https://images.squarespace-cdn.com/content/v1/59cf82d5cd0f6847b8e9572f/1508083466627-WG0O95E5Y9C8KZZH0QGK/neuron_circuit%402x-100.jpg The Team https://www.twelvetreeslab.co.uk/join daily 0.75 2022-09-30 https://www.twelvetreeslab.co.uk/contact daily 0.75 2017-10-16