Cornea. J.J. Clark., 2001. Micron, 32, 239-249. Bloom & Fawcett. 200nm. 250nm
... A text book of histology, Bloom and Fawcett. WHERE ?
The puzzle of the 3D Organization Of the extracellular Matrix of Living Tissues
Gervaise MOSSER Equipe Matériaux du Vivant LCMCP, UMR7574-CNRS-UPMC-EPHE 4 place Jussieu, Case courrier 1220. (Bat. F, 3ème, p332B) 75252 PARIS Cedex 05 00 33 (0)1.44.27.65.53
[email protected] http://www.labos.upmc.fr/lcmcp/newsite/?q=
DIFFERENT TISSUES ANALYSED AT DIFFERENT SCALES
OPTICAL SCALE LOW MAG SEM SCALE Photonique Microscopies : Polarized light
1 μm Section
Second harmonic
Whole mount
Scanning electron microscopy Whole mount
Young Rat Tail Derm.
1μm section, MOP
20μm
Young rat tail tendon.
1μm section, SEM
Nicholls, SP., et al., 1983. Int. J. Biol. Macromol. 5, 283-288.
Young rat tail tendon. Campos Vidal B. (2003). Micron 34, 423-432.
1μm section, MOP
Mouse achille heel tendon. M. Strupler, P.-L. Tharaux et M. –C. Schanne-Klein, LOB, Ecole Polytechnique / CNRS /
SHG-M
1μm section, MOP
Young rat tail bone.
Decalcified Human osteon bone.
M.-M. Giraud-Guille
Rabbit Cornea
Dz=15µm.
20 μm
M. Strupler, M. –C. Schanne-Klein, LOB, Ecole Polytechnique / CNRS /
SHG-M
20 μm
GENERAL FEATURES AT LOW MAGNIFICATIONS TISSUE
ORGANIZATION
Derme
Aleatory « in plane »
Compact Bone
Helicoidal with various degree of helicity depending on origin Contre plaqué helicoidal
Cornea Tendon
Wavy to Straight depending on origin
TRANSMISSION ELECTRON MICROCOPY
Ultra thin sections, 70 - 90 nm
Cells
Extra-cellular Matrix 80 % du poids sec des organismes complexes adultes
F.H. Silver et al. I. Journal of Biomechanics (2003) 36, 1529-1553
0,5 μm
Young Rat Tail Bone. Human demineralized bone. 0,5 μm
Giraud-Guille, M.M., 1996. Int. Rev. Cyto. 166, 59 - 101.
Cornea Bloom & Fawcett
250nm
J.J. Clark., 2001. Micron, 32, 239-249
200nm
MOST STRICKING FEATURE AT HIGH MAGNIFICATIONS
Same major elementary brick D
Individualized cross striated fibrils Variable, but controled diameter
10nm - 200 nm de diamètre, X μm de long
Gap Overlap
Holmes et al. 2001 Process maturation from procollagen to collagen and to fibrils.
N-t O C-t
D
Structural Hierarchical Organization
Os
Monomer left handed helix,
Trimmer right handed
nm
Derme
10-300 nm Fibril Organization of collagen within the fibril ? 1-20 μm Fibre Organization of fibrils within the fibre ? 20-200μm Fasicle Organization of fibres within the fascicle ?
Tendon
WHEN ? WHERE ? WHAT ? HOW ?
Buste d'homme accoudé
Nicolas de Leyde
0 % MEC, 100 %CELL 80% MEC, 20%CELL
WHEN ?
20% MEC, 80%CELL
WHERE ?
A text book of histology, Bloom and Fawcett
F.H. Silver et al. 2003. J. Biomechanics, 36.1529-1553 C
A
D
B
A
C
D
B
WHAT ? Cells Cells tracts on fibrils If the desmosomes formation is inhibited no tendon Kadler, K.
Spacing between fibrils
Silver, F.H.
Diameter of the fibrils
Birk, D.
Explain the fibrils, fibres and fascicles hierarchy BUT nothing published yet that could explain that cells could be responsable for the long range 3D organization
WHAT ? physico – chemistry Cholesteric, Giraud-Guille, 1992 “smectic” Hukins & WoodheadGalloway, 1977 Comper & Veis, 1977 Gobeaux et al. Submitted
Banded texture, Giraud-Guille & Besseau, 1997 Long range organization BUT No published conditions have yet led to individual fibrils and to arrangement into fibers and fascicles in dense solutions
HOW ?
The Cells
The physico – chemistry
Aknowledgments Marie Madeleine GIRAUD-GUILLE (DR, EPHE)
Y. Bouligand K. Kadler Patrick DAVIDSON (LPS, ORSAY) Pierre PANINE (ESRF, ID02) Sylvie HENON (MSC, PARIS) Sophie ASNASCIOS (MSC, PARIS) Uwe HANSEN (Munchen, DE)
Physicochimie Gervaise MOSSER (CR, CNRS) Emmanuel BELAMIE (MC, EPHE►) Frédéric GOBEAUX ( Doctorant ►) Paulo PEIXOTO ( ► Doctorant ) Minéralisation Nadine NASSIF (CR, CNRS) Culture de Cellule Christophe HELARY (AI, EPHE) Sylvain VIGIER ( Doctorant ) Préparation des échantillons MO/MET Anny ANGLO (TE, UPMC) Corinne ILLOUL (AST, UPMC)
La Matrice Extra-Cellulaire 80 % du poids sec des organismes complexes adultes Cohésion des tissus, médium où les cellules sont immergées, nutriments, hormones, enzymes, autres (macro)molecules biologiques, défences (système immunitaire) etc…
famille des collagènes, 1/3 des protéines (29 membres).
JACOB MP
PROCOLLAGENE I COLLAGENE I
Précédent | Accueil | Suivant
http://www.sante.univ-nantes.fr/med/biolcell/matric_extrac/index.htm
Young Rat Tail Tendon.
Young Rat Tail Derm.
Young Rat Tail Bone.
