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Quantitative analysis of cytoskeletal network structures

Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

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Page 1: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Quantitative analysis of

cytoskeletal network structures

Page 2: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

5 µm

MTMF

IF (keratin)

microfilaments(MF)

persistence tensile length [µm] modulus [a.u.]

MF 18 300MT >100 150IF 1 1

intermediatefilaments (IF)

microtubules(MT)

Cytoskeleton

Keratin filaments

Page 3: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Role of cytoskeletal filaments

1. cell and tissue architecture

2. cell migration

Page 4: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Motility of leukaemia cells - vimentin

ATRAPMA

NB4 - Vimentin

neutrophil

Retinoic Acid Syndrome

Page 5: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Tensegrity (R.B. Fuller, D. Ingber)

microtubulesmicrotubules

intermediate filaments

F-actin

substrate anchor

Page 6: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Keratin networks in microgravity

t2µg

Page 7: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

control sphingosylphosphorylcholine (SPC)

Regulation of keratin networks

Page 8: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

min

F-actin

K8/18

Page 9: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Keratin network morphology

perinuclear

periphery

Page 10: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

50 ng/mL TGFα

min

ERK

2 ac

tivat

ion

Seufferlein et al. 1999

control

TGFα (30 min)

Keratin network reorganisation in response to TGFα

Page 11: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility
Page 12: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

control

TGFα, 30 min

Page 13: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

control TGFα control TGFα

Structural features of keratin networks

Page 14: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Fitting of random tessellation models

Poisson-Line (PLT) Poisson-Voronoi (PVT) Poisson-Delaunay (PDT)

Page 15: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Iterated tesselations

Page 16: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Bernoulli thinning

Page 17: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Model selected as best fitcontrol TGFα

Page 18: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

control PVT / PLT

TGFα PDT / PLT

Page 19: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Actin networks in lamellipodia

Page 20: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility
Page 21: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Elasticity ~ mesh size4.4

PVT PDT PLT

Page 22: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

F-actin

Migration

Page 23: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility

Stephanie Eckel, Frank Fleischer, Hendrik Schmidt, Volker SchmidtDept. of Stochastics, University Ulm

Paul WaltherDept. of Electron Microscopy, University Ulm

Thomas SeufferleinDept. of Internal Medicine I, University Ulm

Bishr OmaryDept. of Medicine, Stanford University

Tatjana SvitkinaDept. of Biology, University of Pennsylvania

Joachim SpatzDept. of Physical Chemistry, University Heidelberg

Page 24: Quantitative analysis of cytoskeletal network structures fileCytoskeleton Keratin filaments. Role of cytoskeletal filaments 1. cell and tissue architecture 2. cell migration. Motility