Summary
The binding sites of the following biotinylated lectins were demonstrated in serial paraffin sections of fore- and hindlimb buds from day-9 to day-16 mouse embryos with the Avidin-Biotin-Peroxidase Complex (ABC) procedure: Concanavalin A (Con A), Soybean Agglutinin (SBA), Wheat Germ Agglutinin (WGA), Peanut Agglutinin (PNA), Ricinus Communis Agglutinin I (RCA), Ulex Europaeus I Agglutinin (UEA), and Dolichos Biflorus Agglutinin (DBA). Alternating neighbouring sections were used to compare the distribution of PNA staining, PNA staining after neuraminidase treatment (N-PNA) and the autoradiographic sites of [35S]-sulphate uptake. Unspecific binding sites common to all lectins tested were observed in periderm and chondrocytes. Several lectin affinities were seen in the undifferentiated mesoderm (Con A, WGA, RCA), blood vessels (WGA, PNA, N-PNA, RCA, UEA, DBA) and macrophages (Con A, WGA, N-PNA, RCA). A very selective and mainly extracellular affinity to N-PNA was demonstrated in the condensed preskeletal mesoderm, where it characterizes indistinct prospective chondrogenic, perichondral and pre-articular areas. Comparison with the distribution pattern of [35S]-sulphate uptake and other previously published histochemical data suggests that N-PNA staining occurs at the late blastema stage, i.e. after the stage of cell condensation and before the earliest deposit of stainable matrix in chondrogenic areas. This property later disappears from the chondrifying rudiments, and is maintained in perichondral and pre-articular tissues. Surprisingly, only the pre-articular areas bind PNA without pretreatment with neuraminidase. A transient RCA binding probably related to terminal morphogenesis was detected in the undifferentiated distal part of the predigital columns of day-12 and day-13 limb buds. From the day-13 stage onwards, diverse new lectin affinities appeared in differentiating tissues, such as pretendinous rudiments, perichondrium and prospective periosteum, muscular connective tissue, myotubes, superficial fasciae and prospective dermis. A strong SBA and PNA staining was also detected in the extracellular matrix associated with the epithelial septa separating the roots of the digits in day-15 and day-16 limb buds.
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References
Aulthouse AL, Solursh M (1987) The detection of a precartilage blastema-specific marker. Dev Biol 120:377–384
Caterson B, Mahmoodian F, Sorrell JM, Hardingham TE, Bayliss MT, CArney SL, Ratcliffe A, Muir H (1990) Modulation of native chondroitin sulphate structure in tissue development and in disease. J Cell Sci 97:411–417
Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92
Hewitt AT, Elmer WA (1976) Reactivity of normal and brachypod mouse limb mesenchymal cells with Con-A. Nature 264:177–178
Hewitt AT, Elmer WA (1978) Developmental modulation of lectinbinding sites on the surface membrane of normal and brachypod mouse limb mesenchymal cells. Differentiation 10:31–38
Hsu SM, Raine L, Fanger H (1981) Use of Avidin-Biotin-Peroxidase Complex (ABC) and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
Hurle JM, Lafarga M, Hinchliffe JR (1981) The surface coat of embryonic limb mesenchymal cells during morphogenetic cell death. An ultrastructural study of chick interdigital necrotic zones (INZ) using ruthenium red and concanavalin A. Exp Cell Res 133:465–470
Hurle JM, Ros MA, Hinchliffe JR (1988) Spatial and temporal changes in the pattern of glycosylation of the developing chick limb tissue components as revealed by fluorescent conjugated lectin probes. Cell Differ 24:149–158
Hurle JM, Ros MA, Ganan Y, Macias D, Critchlow M, Hinchliffe JR (1990) Experimental analysis of the role of ECM in the patterning of the distal tendons of the developing limb bud. Cell Differ 30:97–108
Lison L (1953) Histochimie et cytochimie animales. Gauthiers Villars, Paris
Mallinger R, Geleff S, Bock P (1986) Histochemistry of glycosaminoglycans in cartilage ground substance. Alcian-blue staining and lectin-binding affinities in semithin Epon sections. Histochemistry 85:121–127
Milaire J (1965) Etude morphogénétique de trois malformations congénitales de l'autopode chez la souris (syndactylisme - brachypodisme - hémimélic dominante) par des méthodes cytochimiques. Acad R Belg Cl Sci Mém in 4° (Sc.) 16:1–120
Milaire J (1969) Etude morphogénétique de la syndactylie postaxiale provoquée chez le rat par l'hadacidine. I. Analyse des anomalies chez l'adulte, le foetus a terme et les embryons de 15, 16 et 17 jours. Arch Biol 80:167–253
Milaire J (1978) Etude morphologique, histochimique et autoradiographique du développement du squelette des membres chez l'embryon de souris. I. Membres antérieurs. Arch Biol 89:169–216
Milaire J (1983) Patterns of dephosphorylating activities in the mesoderm of developing mouse limb buds. I. 5′Nucleotidase, non specific ATP-phosphohydrolase and alkaline phosphatase in normal forelimb buds. Arch Biol 94:301–344
Milaire J (1991) Lectin histochemistry in normal and abnormal limb morphogenesis in the mouse. Progr Histochem Cytochem 23 (in press)
Milaire J, Rooze M (1982) Etude morphologique, histochimique et autoradiographique du développement du squelette des membres chez l'embryon de souris. II. Membres postérieurs. Arch Biol 93:311–343
Milaire J, Rooze M (1983) Hereditary and induced modifications of the normal necrotic patterns in the developing limb buds of the rat and mouse: facts and hypotheses. Arch Biol 94:459–490
Paulsen DF, Finch RA, Parker CL (1980) Age- and region-dependent redistribution of Con A binding sites on chick wing bud mesoderm cells. J Exp Zool 213:369–376
Welim HB, Thies M, Herken R (1989) Appearance of lectin-binding sites during vascularization of the primordium of the central nervous system in 10 to 12-day-old mouse embryos. Cell Tissue Res 255:627–630
Yamagata T, Nishiwaki T (1979) Chick embryonic cartilage has a developmentally regulated lectin specific for galactose-containing saccharides. Proc Japan Acad 55(Ser.B):393–397
Zimmermann B (1986) Binding of various lectins during chondrogenesis in mouse limb buds. Acta Histochem 32s:127–131
Zimmermann B, Thies M (1984) Alterations of lectin binding during chondrogenesis of mouse limb buds. Histochemistry 81:353–361
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Milaire, J. Lectin binding sites in developing mouse limb buds. Anat Embryol 184, 479–488 (1991). https://doi.org/10.1007/BF01236054
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DOI: https://doi.org/10.1007/BF01236054