Intranuclear distribution of galectin-3 in mouse 3T3 fibroblasts: comparative analyses by immunofluorescence and immunoelectron microscopy.
The intracellular distribution of carbohydrate binding protein 35 (CBP35), recently named galectin-3, was studied in mouse 3T3 fibroblasts, using immunofluorescence at the light microscope level and immunogold labeling at the ultrastructural level. In general, serum-stimulated, proliferating cells showed higher levels of labeling than quiescent cultures of the same cells. In the proliferating cells, the labeling intensity was higher in the nucleus than in the cytoplasm. Treatment of permeabilized cells or thin sections with ribonuclease A decreased the immunolabeling intensity, whereas parallel control treatments with deoxyribonuclease I failed to yield the same effect. While there appears to be general agreement between the immunofluorescence and the ultrastructural results regarding the level of CBP35 and its association with nuclear ribonucleoprotein complexes, there was one striking difference in terms of labeling of specific subnuclear structures. Immunofluorescence results indicate diffuse distribution of CBP35 within the nucleus, but the label appears to be excluded from certain "black holes," which most probably correspond to nucleoli. On the other hand, immunogold particles were observed in electron microscopy, mainly in interchromatin spaces, except for interchromatin granule clusters, at the border of condensed chromatin, on the dense fibrillar component, and at the periphery of the fibrillar centers of nucleoli.
Hubert
M
M
Laboratoire de Glycobiologie et de Reconnaissance Cellulaire, U180 INSERM, UFR Biomédicale des Saints-Pères (Université René Descartes, Paris V), France.
Wang
S Y
SY
Wang
J L
JL
Sève
A P
AP
Hubert
J
J
D016475:3T3 Cells; D000818:Animals; D000943:Antigens, Differentiation / Q000032:analysis*; D002466:Cell Nucleolus / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure*; D002843:Chromatin / Q000648:ultrastructure; D016895:Culture Media, Serum-Free; D003593:Cytoplasm / Q000648:ultrastructure; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D037502:Galectin 3; D008562:Membrane Glycoproteins / Q000032:analysis*; D051379:Mice; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D012259:Ribonuclease, Pancreatic
D000943:Antigens, Differentiation; D002843:Chromatin; D016895:Culture Media, Serum-Free; D005452:Fluoresceins; D005456:Fluorescent Dyes; D037502:Galectin 3; D008562:Membrane Glycoproteins; D012259:Ribonuclease, Pancreatic