| Relationship between cell differentiation and binding of fluorescein isothiocyanate (FITC)-conjugated insulin of keratinocytes in culture. | |
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MedLine Citation:
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PMID: 2439607 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Basal-type keratinocytes, isolated from newborn rat skin and separated on Percoll density gradients, proliferate in low (0.1 mM) calcium medium and, after raising the calcium level to normal (1.96 mM), stratify. Cells in the low calcium culture do not have extensive cell-cell connections, as seen with fluorescein isothiocyanate (FITC)-conjugated insulin. Fluorescein isothiocyanate-conjugated concanavalin A and Griffonia simplicifolia isolectin B4, but not peanut agglutinin (PNA), fluorescently label these cells. In 3-day-old low calcium cultures, within 2 h after raising the calcium of the medium to the normal level, intense binding of PNA to cells appears and neighboring cells are connected through bundles of filaments that are fluorescently labeled by FITC-insulin. After 2 days in normal calcium medium, the cultures exhibit relatively smooth, straightlined, cell boundaries that are labeled by FITC-insulin and cell boundaries and intracellular granules that are stained by hematoxylin. One day later, similar cell boundaries are present, but they are not significantly decorated by FITC-insulin and, under phase contrast microscopy, are dark. Free FITC gives labeling patterns similar to those given by FITC-insulin, but the FITC labeling is blocked by mercaptoethanol and dithiothreitol in contrast to FITC-insulin binding. The present results suggest the insulin moiety is involved in the labeling by FITC-insulin and the labeling is chronologically related to the stage of cell differentiation. |
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Authors:
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H W Feng; X P Jin; I A Bernstein |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: The Journal of investigative dermatology Volume: 89 ISSN: 0022-202X ISO Abbreviation: J. Invest. Dermatol. Publication Date: 1987 Jul |
Date Detail:
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Created Date: 1987-07-28 Completed Date: 1987-07-28 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0426720 Medline TA: J Invest Dermatol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 73-7 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Calcium / pharmacology Cell Differentiation Cells, Cultured Concanavalin A / analogs & derivatives, metabolism Culture Media Epidermis / cytology*, metabolism Fluorescein-5-isothiocyanate* / analogs & derivatives* Fluoresceins / metabolism Insulin / analogs & derivatives*, metabolism, pharmacology Insulin Antagonists Keratins* Lectins / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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AM07236/AM/NIADDK NIH HHS; AM15206/AM/NIADDK NIH HHS; CA32470/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Culture Media; 0/Fluoresceins; 0/Insulin Antagonists; 0/Lectins; 0/fluorescein isothiocyanate-concanavalin A; 0/insulin, fluorescein-isothiocyanated-; 11028-71-0/Concanavalin A; 11061-68-0/Insulin; 3326-32-7/Fluorescein-5-isothiocyanate; 68238-35-7/Keratins; 7440-70-2/Calcium |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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