The effects of 30- and 45-minute trypsin- and pancreatin-separation on embryonic rabbit palate epithelium and mesenchyme were studied with the electron microscope. Changes in epithelium included fragmentation of the basal lamina and formation of cytoplasmic blebs associated with the basal cell layer. Changes in mesenchymal cells were first evident at 45 min of incubation and included cell fragmentation, nuclear pyknosis, and dilation of extracellular space. Results indicate a differential susceptibility of mesenchyme to the separating agent.
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References
1.
DePaola, D.P. ; Miller, S.A.; and Drummond , J.F.: The Biochemical Growth and Development of the Palate, Maxilla, Mandible, and Tongue in the New Zealand White Rabbit, Arch Oral Biol18:1113-1119, 1973.
2.
Lessard, J.L. ; Wee, S.L.; and Zimmer-Man , S.F.: Presence of Contractile Proteins in Mouse Fetal Palate Prior to Shelf Elevation, Teratology9:113-125, 1974.
3.
Pratt, R.M.; Goggins, J.F.; Wilk, A.L.; and King, C.T.: Acid Mucopolysacchatide Synthesis in the Secondary Palate of the Developing Rat at the Time of Rotation and Fusion, Dev Biol32:230-237,1973.
4.
Larsson, K.: Occurrence of Sulfomucopolysaccharides in the Palatine Processes of the Mouse Embryo, Exp Cell Res21 :498-503, 1960.
5.
WALKER, B.E.: The Association of Mucopolysaccharides with Morphogenesis of Palatal and Other Structures in Mouse Embryos, J Embryol Exp Morphol 9:22-31, 1961. 6. JACOBS, R.: S35-liquid Scintillation Count Analysis of Morphogenesis and Teratogenesis of the Palate in Mouse Embryos, Anat Rec 150:271-278, 1964.
6.
DePaola, D.P. ; Lorente, C.L.; Drummond , J.F.; Miller, S.A.; and Zarbo, R.: Glycoprotein Biosynthesis at the Time of Palate Fusion by Rabbit Palate and Maxilla Cultured in vitro, J Dent Res54:1049-1055, 1975 .
7.
Pratt, R.M. and Hassell, J.R.: Appearance and Distribution of Carbohydrate-rich Macromolecules on the Epithelial Surface of the Developing Rat Palatal Shelf, Dev Biol45: 192-198,1975.
8.
Souchon, R.: Surface Coat of the Palatal Shelf Epithelium During Palatogenesis in Mouse Embryos, Anat Embryol147:133-142. 1975,
9.
Pratt, R.M. and King, C.T.G.: Inhibition of Collagen Cross-linking Associated with β-aminopioprionitrile-induced Cleft Palate in the Rat , Dev Biol27:322-328, 1972.
10.
Pourtois, M. : Onset of the Acquired Potentiality for Fusion in the Palatal Shelves of Rats, J Embryol Exp Morphol16:171-182, 1966.
11.
Pourtois, M. : The Fate of Rat Palatal Shelves Cultivated in the Presence of Periodic Acid, Arch Oral Biol16:503-508, 1970.
12.
DePaola, D.P. ; Drummond, J.F.; and Miller, S.A.: An Autoradiographic Study Demonstrating 3H-fucose Uptake into Cultured Palates at the Time of Palatal Fusion, J Dent Res54:912,1975.
13.
Greene, R.M. and Pratt, R.M.: Inhibition by Diazo-oxo-norleucine (DON) of Rat Palatal Glycoprotein Synthesis and Epithelial Cell Adhesion in vitro , Exp Cell Res105: 27-37, 1977.
14.
Neiders, M.E. and Weiss, L.: Isolation of Viable Epithelial Cells from Human Gingiva, J Periodontol41:325-332, 1970.
15.
Einbinder, J.M. ; Walzer, R.A.; and Mandel, I.: Epidermal-dermal Separation with Proteolytic Enzymes, J Invest Derm46: 492-504,1966.
16.
Grobstein, C. : Epithelio-mesenchymal Specificity in the Morphogenesis of Mouse Submandibular Rudiments in vitro, J Exp Zool124:383-413, 1953.
17.
Scaletta, L.J. and McCallum, D.K.: Divalent Cation-mediated Epithelial Union with Connective Tissue, Exp Cell Res68: 449-452, 1971.
18.
Baumberger, J.P.; Suntzeff, V.; and Condry, E.V.: Methods for the Separation of Epidermis From Dermis and Some Physiologic and Chemical Properties of Isolated Epidermis , J Natl Cancer Inst2:413-423, 1942.
19.
Auerbach, R. and Grobstein, C.: Inductive Interaction of Embryonic Tissues After Dissociation and Reaggregation, Exp Cell Res15:384-397, 1958.
20.
Tyler, M.S.: Epithelial-mesenchymal Interactions in the Embryonic Secondary Palate of the Mouse: An in vitro Study, Ph.D. Dissertation, University of North Carolina, 1975.
21.
Hudson, C.D. and Shapiro, B.L.: A Radio-autographic Study of Deoxyribonucleic Acid Synthesis in Embryonic Rat Palatal Shelf Epithelium with Reference to the Concept of Programmed Cell Death, Arch Oral Biol18: 77-84, 1973.
22.
Pratt, R.M. and Martin, G.R.: Epithelial Cell Death and Cyclic AMP Increase During Palatal Development, Proc Natl Acad Sci72: 874-877,1975.
23.
Tyler, M.S. and Koch, W.E.: Epithelial-mesenchymal Interactions in the Secondary Palate of the Mouse, J Dent Res53:64, 1974.
24.
Scaletta, L.J. and McCallum, D.K.: A Fine Structural Study of Human Oral Epithelium Separated from the Lamina Propria by Enzymatic Action, Am J Anat140:383-404, 1974.
25.
Kallman, F. ; Evans, J.; and Wessels, N.K.: Normal Epidermal Basal Cell Behavior in the Absence of Basement Membrane, J Cell Biol32:231-236, 1967.
26.
Slavkin, H.C. ; Bringas, P.; Lebaron , R.; Cameron, J.; and Bavetta, L.A.: The Fine Structure of the Extracellular Matrix During Epithelio-mesenchymal Interactions in the Rabbit Embryonic Incisor , Anat Rec165:237-256,1969.
27.
Goel, S.C. and Jurand, A.: Electron Microscopic Observations on the Basal Lamina of Chick Limb Buds After Trypsin and EDTA Treatment, J Cell Sci3:373-380, 1968.
28.
Dodson, J.W. : The Differentiation of Epidermis. I. The Interrelationships of Epidermis and Dermis in Embryonic Chicken Skin, J Embryol Exp Morphol17:83-106, 1967.
29.
Jensen, H.M. and Mottet, N.K.: Ultrastructural Changes in Keratinizing Epithelium Following Trypsinization, Epidermal Detachment, and Apposition to Mesenchymes, J Cell Sci6:511-535, 1970.
30.
Briggman, R.A. ; Daldorf, F.C.; and Wheller, C.E.: Formation and Origin of Basal Lamina and Anchoring Fibrils in Adult Human Skin, J Cell Biol51:384-395, 1971.
31.
Smiley, G.R. and Koch, W.E.: The Fine Structure of Mouse Secondary Palate Development in vitro, J Dent Res50:1671-1677, 1971.
32.
Caesar, R. and Edwards, G.A.: The Physiologic Significance of the Basement Membrane, Anat Rec128:530, 1957.
33.
Pourtois, M. : Morphogenesis of Primary and Secondary Palate . In: Developmental Aspects of Oral Biology, H. Slavkin and L. Bavetta , Eds., New York : Academic Press , 1973, pp. 80-108.
34.
Felton, J.H. ; Person, P.; and Stahl, S.S.: Biochemical and Histochemical Studies of Aerobic Oxidative Metabolism of Oral Tissues. II. Enzymatic Dissection of Gingival and Tongue Epithelia from Connective Tissues, J Dent Res44:392-401, 1965.
35.
Tsanev, R.G. and Markov, G.G.: Nucleic Acid Content in Mouse Epidermis Separated from Dermis by Different Methods, Experientia16:448-449, 1960.
36.
Moscona, A.A. : Rotation-mediated Histogenetic Aggregation of Dissociated Cells: A Quantifiable Approach to Cell Interactions in vitro , Exp Cell Res22:455-475, 1961.
37.
Moscona, A.A. and Moscona, M.H.: Comparison of Aggregation of Embryonic Cells Dissociated with Trypsin or Versene, Exp Cell Res45:239, 1967.
38.
Tyler, M.S. and Koch, W.E.: In vitro Development of Palatal Tissues from Embryonic Mice. II. Tissue Isolation and Recombination Studies , J Embryol Exp Morphol38 :19-36, 1977.
