Summary
The effects of collagenase on the immunohistochemical demonstrability of laminin, fibronectin and Factor VIII/RAg in human nervous tissue have been studied. The influence of this, and other proteolytic enzymes such as pepsin and trypsin, has been investigated in relation to different fixatives. Collagenase gave better results with Carnoy fixed material than after formalin fixation; unlike trypsin and pepsin, it did not produce tissue digestion.
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Bloom AL, Giddings JC, Wilks CJ (1973) F VIII on the vascular intima: possible importance in haemostasis and thrombosis. Nature (New Biol) 241:217–219
Brozman M (1978) Immunohistochemical analysis of formaldehyde-and trypsin or pepsin-treated material. Acta Histochem 63:251–260
Burns J, Dixon AJ, Woods JC (1980) Immunoperoxidase localization of fibronectin in glomeruli of formalin fixed paraffin processed renal tissue. Histochemistry 67:73–78
Curran RC, Gregory J (1977) The unmasking of antigens in paraffin sections of tissue by trypsin. Experientia 33:1400–1401
Curran RC, Gregory J (1978) Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques. Effects of processing on immunohistochemical performance of tissues and on the use of proteolytic enzymes to unmask antigens in sections. J Clin Pathol 31:974–983
De Lellis RA (1981) Basic techniques of immunohistochemistry. In: De Lellis RA (ed) Diagnostic immunohistochemistry. Masson, New York
Del Rosso M, Cappelletti R, Viti M, Vannucchi S, Chiarugi V (1981) Binding of the basement-membrane glycoprotein laminin to glycosaminoglycans. Biochem J 199:001–005
Dessau W, Adelman BC, Timpl R, Martin GR (1978) Identification of the sites in collagen α-chains that bind serum anti-gelatin factor (cold-insoluble globulin). Biochem J 169:55–59
Dixon AJ, Burns J, Dunnhill MS, McGee J (1980) Distribution of fibronectin in normal and diseased human kidney. J Clin Pathol 33:1021–1028
Ekblom P, Miettinen M, Rapola J, Foidart JM (1982) Demonstration of laminin, a basement membrane glycoprotein, in routinely processed formalin-fixed human tissues. Histochemistry 75:301–307
Foidart JM, Bere EW, Yaar M, Rennard S, Gullino M, Martin GR (1980) Distribution and immunoelectron microscopic localization of laminin, a non-collagenous basement membrane glycoprotein. Lab Invest 42:336–342
Hay ED (1981) Extracellular matrix. J Cell Biol 91:205s-223s
Hautzer NW, Wittkuhn JF, Mc Caughey WT (1980) Trypsin digestion in immunoperoxydase staining. J Histochem Cytochem 28:52–53
Heyderman E (1979) Immunoperoxidase technique in histopathology; application, methods and controls. J Clin Pathol 32:971–978
Hølund B, Clemmensen I (1982) The value of hyaluronidase treatment of different tissues before demonstration of fibronectin by the indirect immunoperoxidase technique. Histochemistry 76:517–525
Hølund B, Clausen PP, Clemmensen I (1981) The influence of fixation and tissue preparation on the immunohistochemical demonstration of fibronectin in human tissue. Histochemistry 72:291–299
Hynes RO, Yamada KM (1982) Fibronectins: multifunctional modular glycoproteins. J Cell Biol 95:369–377
Kleinman HK, Mc Goodwin EB, Martin GR, Klebe RJ, Fietzek PP, Wolley DE (1978) Localization of the binding site for cell attachment in the 1(I) chain of collagen. J Biol Chem 253:5642–5646
Kochi N, Tani E, Morimura T, Itagaki T (1983) Immunohistochemical study of fibronectin in human glioma and meningioma. Acta Neuropathol 59:119–126
Jones TR, Ruoslahti E, Schold SC, Bigner DD (1982) Fibronectin and glial fibrillary acidic protein expression in normal brain and anaplastic human gliomas. Cancer Res 42:168–177
Laurie GW, Leblond CP, Martin GR (1982) Localization of type IV collagen, laminin, heparan sulfate proteoglycan, and fibronectin to the basal lamina of basement membranes. J Cell Biol 95:340–344
McComb RD, Trevor RJ, Pizzo SV, Bigner DD (1982a) Specificity and sensitivity of immunohistochemical detection of F VIII/RAg in formalin-fixed paraffin-embedded tissue. J Histochem Cytochem 30:371–377
McComb RD, Jones TR, Pizzo SV, Bigner DD (1982b) Immunohistochemical detection of F VIII/von Willebrand factor in hyperplastic endothelial cells in glioblastoma multiforme and mixed glioma-sarcoma. J Neuropathol Exp Neurol 41:479–489
Mepham BL (1982) Influence of fixatives on the immunoreactivity of paraffin sections. Histochem J 14:731–737
Mepham BL, Frater W, Mitchell BS (1979) The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique. Histochem J 11:345–357
Mukai K, Rosai J, Burgdorf WHC (1980) Localization of F VIII/RAg in vascular endothelial cells using an immunoperoxidase method. Am J Surg Pathol 4:273–276
Paetau A, Mellström K, Westermark B, Dahl D, Haltia M, Vaheri A (1980) Mutually exclusive expression of fibronectin and glial fibrillary acid protein in cultured brain cells. Exp Cell Res 129:337–344
Pearse AGE (1968) Histochemistry. Theoretical and applied. Churchill, London
Peterkofsky B (1982) Bacterial collagenase. In: Cunningham LW, Frederiksen DW (eds) Methods in enzymology, vol 82: Academic Press, New York, pp 453–471
Peters A, Palay SL, Webster HF (1976) The fine structure of the nervous system. Saunders, Philadelphia
Reading M (1977) A digestion technique for the reduction of background staining in the immunoperoxidase method. J Clin Pathol 30:88–90
Reitamo S (1978) Lysozyme antigenicity and tissue fixation. Histochemistry 55:197–207
Ruoslahti E, Engvall E, Hayman EG (1981) Fibronectin: current concepts of its structure and function. Cell Res 1:95–128
Ruoslahti E, Hayman G, Pierschbacher M, Engvall E (1982) Fibronection: purification, immunochemical properties and biological activities. In: Cunningham LW, Frederiksen DW (eds) Methods in enzymology, vol 82. Academic Press, New York, 803–831
Sabatini DS, Bensch K, Barnett RJ (1963) Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol 17:19–58
Sanes JR (1982) Laminin, fibronectin and collagen in synaptic and extrasynaptic portions of muscle fiber basement membrane. J Cell Biol 93:442–451
Sehested M, Hou-Jensen K (1981) Factor VIII related antigen as an endothelial cell marker in benign and malignant diseases. Virchows Arch (Pathol Anat) 391:217–224
Seifter S, Harper E (1970) Collagenases. In: Perlmann GE, Lorand L (eds) Methods in enzymology, vol 19. Academic Press, New York, pp 613–635
Sternberger LA, Hardy PH, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method for immunohistochemistry. Preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333
Tarone G, Ceschi P, Prat M, Comoglio PM (1981) Transformation-sensitive protein with molecular weight of 45,000 secreted by mouse fibroblasts. Cancer Res 41:3648–3652
Timpl R, Rohde H, Robey PG, Rennard SI, Foidart JM, Martin GR (1979) Laminin. A glycoprotein from basement membranes. J Biol Chem 254:9933–9937
Timpl R, Rohde H, Risteli L, Ott U, Robey PG, Martin GR (1982) Laminin. In: Cunningham LW, Frederiksen DW (eds) Methods in enzymology, vol 82. Academic Press, New York, 831–838
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Partially supported by CNR Special Project “Control of Neoplastic Growth”, Grant No 82.00410.96
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Mauro, A., Bertolotto, A., Germano, I. et al. Collagenase in the immunohistochemical demonstration of laminin, fibronectin and factor VII/RAg in nervous tissue after fixation. Histochemistry 80, 157–163 (1984). https://doi.org/10.1007/BF00679991
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DOI: https://doi.org/10.1007/BF00679991