title: Nuclear and cytoplasmic lectin receptor sites in rat Py1a osteoblasts.
abstract: The intracellular distribution of lectin receptor sites was studied in the rat Pyla osteoblasts using immunofluorescence at the confocal microscopy level. This immortalized cell line was found to represent a satisfactory model to study the occurrence and distribution of sugar moieties. Our data showed distinct affinity patterns of lectins recognizing different terminal or internal sugar residues. For some lectins, the binding patterns appeared to be cell cycle-independent, whereas for PNA the cell cycle greatly influenced the nuclear binding. By combining lectin affinity with sialidase degradation and alcoholic saponification the sialic acid acceptor sugars and derivatives were also visualized. In particular, glycoconjugates with sialic acids linked to beta-galactose, and mainly C4 acetylated, were located in the cytoplasm, while glycoconjugates characterized by sialic acids linked to alpha-N-acetylgalactosamine, and devoid of acetyl groups at C4, were almost exclusively found in the nucleus. The comparison of lectin affinities, with and without prior glycosidase digestions, allowed us to gain further insight into the chemical composition of glycoconjugates that act as the lectin receptor sites that appeared to belong to O- and N-linked glycoconjugates. The use of additional enzymatic treatments were useful to better establish the localization of nuclear receptor sites and results were compared with previous studies about endogenous and exogenous lectins in an attempt to reconcile the association of lectins and sugars within the nucleus and their possible involvement in modulation of cell proliferation and/or response to chemical signals. The above digestions also provided information about the cytoplasmic binding patterns.
authors.affiliation: Department of Comparative Morphological and Biochemical Sciences, University of Camerino, Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Sabbieti
authors.forename: L
authors.initials: L
authors.lastname: Marchetti
authors.forename: M H
authors.initials: MH
authors.lastname: Hurley
authors.forename: G
authors.initials: G
authors.lastname: Menghi
mesh_terms: D000645:Ammonium Sulfate; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D017364:Collagenases; D003593:Cytoplasm / Q000378:metabolism*; D003640:Dealkylation; D003851:Deoxyribonucleases; D006868:Hydrolysis; D037102:Lectins; D018613:Microscopy, Confocal; D009439:Neuraminidase / Q000737:chemistry; D010006:Osteoblasts / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D011975:Receptors, Mitogen / Q000378:metabolism*; D012260:Ribonucleases; D016707:Tissue Fixation; D001616:beta-Galactosidase
chemical_list: D037102:Lectins; D011975:Receptors, Mitogen; D003851:Deoxyribonucleases; D012260:Ribonucleases; D009439:Neuraminidase; D001616:beta-Galactosidase; D017364:Collagenases; D000645:Ammonium Sulfate