Summary
The ultrastructure of the collagen of rat tail tendon was investigated by the freeze-fracture technique. Collagen fibers were pretreated with the digestive enzymes, α-amylase, elastase and collagenase to remove matrix substances. Some of the samples were etched for 20 min. Fibrils had an average diameter of 318±12 nm and a banded structure with a mean periodicity of 64.2±0.9 mm; the banding was most marked in α-amylase/elastase-treated specimens, although the periodicity was independent of pretreatment. Microfibrils were well-displayed following α-amylase/elastase and collagenase pretreatments. A difference in the diameters of microfibrils was, however, observed between etched specimens (8.3±0.3 nm) and those prepared by other experimental methods (11.4±0.5 nm). In replicas of collagenase-treated and etched specimens, the interconnecting filaments in the interfibrillar region formed a network that was continuous with the microfibrils of collagen fibrils. The diameter of the interconnecting filaments was the same as that of microfibrils. Microfibrillar bundles were observed in the interfibrillar region.
Similar content being viewed by others
References
Belton JC, Michaeli D, Fudenberg HH (1975) Freeze-etch study of collagen. I. Native collagen from tendon and lung of rats. Arthritis Rheumat 18:443–450
Bouteille M, Pease DC (1971) The tridimensional structure of native collagenous fibrils, their proteinaceous filaments. J Ultrastruct Res 35:314–338
Bruns RR (1976) Supramolecular structure of polymorphic collagen fibrils. J Cell Biol 68:521–538
Bruns RR, Trelstad RL, Gross J (1973) Cartilage collagen: A staggered substructure in reconstituted fibrils. Science 181:269–271
Castellani PP, Morocutti M, Franchi M, Ruggeri A, Bigi A, Roveri N (1983) Arrangement of microfibrils in collagen fibrils of tendons in the rat tail. Ultrastructural and X-ray diffraction investigation. Cell Tissue Res 234:735–743
Chapman JA (1974) The staining pattern of collagen fibrils I. An analysis of electron micrographs. Connect Tissue Res 2:137–150
Chapman JA, Hulmes DJS (1984) Electron microscopy of the collagen fibril. In: Ruggeri A, Motta PM (eds) Ultrastructure of the connective tissue matrix. Martinus Nijhoff Publishers, Boston, pp 1–33
Chapman JA, Holmes DF, Meek KM, Rattew CJ (1981) Electronoptical studies of collagen fibril assembly. In: Balaban M, Sussman JL, Traub W, Yonath A (eds) Structural aspects of recognition and assembly in biological macromolecules. Balaban ISS, Rehovot, pp 387–401
Doyle BB, Hulmes DJS, Miller A, Parry DAD, Piez KA, Woodhead-Galloway J (1974) A D-periodic narrow filament in collagen. Proc R Soc Lond B 186:67–74
Gotoh T, Sugi Y, Hirakow R (1983) Ultrastructural construction of collagen fibrils as revealed by the freeze-fracture technique. J Electron Microsc 32:213–215
Hodge AJ, Petruska JA (1963) Recent studies with the electron microscope on ordered aggregates of the tropocollagen macromolecule. In: Ramachandran GN (ed) Aspects of protein structure. Academic Press, New York, pp 289–300
Hulmes DJS, Miller A, White SW, Timmins PA, Berthet-Colominas C (1980) Interpretation of the low-angle meridional neutron diffraction patterns from collagen fibres in terms of the amino acid sequence. Int J Biol Macromol 2:338–346
Hulmes DJS, Jesior JC, Miller A, Berthet-Colominas C, Wolff C (1981) Electron microscopy shows periodic structure in collagen fibril cross sections. Proc Natl Acad Sci USA 78:3567–3571
Itoh Y, Klein L, Geil PH (1982) Age dependence of collagen fibril and subfibril diameters revealed by transverse freeze-fracture and -etching technique. J Microsc 125:343–357
Junqueira LCU, Montes GS (1983) Biology of collagen-proteoglycan interaction. Arch Histol Jpn 46:589–629
Kajikawa K, Nakanishi I, Hori I, Matsuda Y, Kondo K (1970) Electron microscopic observations on connective tissues using ruthenium red staining. J Electron Microsc 19:347–354
Lillie JH, MacCallum DK, Scaletta LJ, Occhino JC (1977) Collagen structure: Evidence for a helicoidal organization of the collagen fibril. J Ultrastruct Res 58:134–143
Marchini M, Ruggeri A (1984) Ultrastructural aspects of freezeetched collagen fibrils. In: Ruggeri A, Motta PM (eds) Ultrastructure of the connective tissue matrix. Martinus Nijhoff Publishers, Boston, pp 89–94
Meek KM, Chapman JA, Hardcastle RA (1979) The staining pattern of collagen fibrils. Improved correlation with sequence data. J Biol Chem 254:10710–10714
Myers DB, Highton TC, Rayns DG (1969) Acid mucopolysaccharides closely associated with collagen fibrils in normal human synovium. J Ultrastruct Res 28:203–213
Myers DB, Highton TC, Rayns DG (1973) Ruthenium red-positive filaments interconnecting collagen fibrils. J Ultrastruct Res 42:87–92
Pease DC, Bouteille M (1971) The tridimensional ultrastructure of native collagenous fibrils, cytochemical evidence for a carbohydrate matrix. J Ultrastruct Res 35:339–358
Rayns DG (1974) Collagen from frozen fractured glycerinated beef heart. J Ultrastruct Res 48:59–66
Ruggeri A, Benazzo F, Reale E (1979) Collagen fibrils with straight and helicoidal microfibrils: A freeze-fracture and thin-section study. J Ultrastruct Res 68:101–108
Smith JW (1968) Molecular pattern in native collagen. Nature 219:157–158
Stolinski C, Breathnach AS (1977) Freeze-fracture replication and surface sublimation of frozen collagen fibrils. J Cell Sci 23:325–334
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gotoh, T., Sugi, Y. Electron-microscopic study of the collagen fibrils of the rat tail tendon as revealed by freeze-fracture and freeze-etching techniques. Cell Tissue Res. 240, 529–534 (1985). https://doi.org/10.1007/BF00216341
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00216341