Visualization of glycosaminoglycans in rat incisor predentin and dentin with cetylpyridinium chloride-glutaraldehyde as fixative.
Using cetylpyridinium chloride (CPC) in glutaraldehyde as fixative, we observed sinuous fiber-like structures 300-500 nm long and 7-14 nm thick in the spaces between the collagen fibers of rat incisor predentin. Small granules and fibrils were also detected. Electron-dense vesicles were seen inside the odontoblast processes. The plasma membrane was irregularly stained with material that adhered to its surface. In demineralized dentin, needle-like structures were seen at the periphery of globular structures which were not stained. Staining the sections with Alcian blue did not greatly improve the visualization of CPC-precipitated glycosaminoglycans. The specificity of staining was assessed on serial sections by selective dissociation of glycosaminoglycan aggregates with 2 M calcium chloride and their digestion by bovine testicular hyaluronidase. The glycosaminoglycans were probably combined with lipids, because treatment of the sections with a chloroform/methanol mixture removed the CPC-induced precipitates from both predentin and dentin.
Chardin
H
H
Laboratoire d'Histologie, Faculté de Chirurgie Dentaire, Montrouge, France.
Septier
D
D
Goldberg
M
M
D000447:Aldehydes; D000818:Animals; D002122:Calcium Chloride; D002462:Cell Membrane / Q000648:ultrastructure; D002594:Cetylpyridinium; D011232:Chemical Precipitation; D002725:Chloroform; D003804:Dentin / Q000032:analysis*; D003810:Dentinogenesis / Q000502:physiology*; D005404:Fixatives; D005976:Glutaral; D006025:Glycosaminoglycans / Q000032:analysis*; D006821:Hyaluronoglucosaminidase; D007180:Incisor / Q000648:ultrastructure; D008854:Microscopy, Electron / Q000379:methods; D009804:Odontoblasts / Q000032:analysis / Q000648:ultrastructure; D011726:Pyridinium Compounds; D051381:Rats; D011919:Rats, Inbred Strains
D000447:Aldehydes; D005404:Fixatives; D006025:Glycosaminoglycans; D011726:Pyridinium Compounds; D002725:Chloroform; D002594:Cetylpyridinium; D006821:Hyaluronoglucosaminidase; D002122:Calcium Chloride; D005976:Glutaral