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The adhesion force of Notch with Delta and the rate of Notch signaling.

Ahimou F, Mok LP, Bardot B, Wesley C - J. Cell Biol. (2004)

Bottom Line: Notch signaling is repeatedly used during animal development to specify cell fates.Reduced turnover or Delta pulling accelerate this loss.These data suggest that strong adhesion between Notch and Delta might serve as a booster for initiating Notch signaling at a high rate.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Molecular Genetics, The University of Vermont, VT 05405, USA.

ABSTRACT
Notch signaling is repeatedly used during animal development to specify cell fates. Using atomic force microscopy on live cells, chemical inhibitors, and conventional analyses, we show that the rate of Notch signaling is linked to the adhesion force between cells expressing Notch receptors and Delta ligand. Both the Notch extracellular and intracellular domains are required for the high adhesion force with Delta. This high adhesion force is lost within minutes, primarily due to the action of Presenilin on Notch. Reduced turnover or Delta pulling accelerate this loss. These data suggest that strong adhesion between Notch and Delta might serve as a booster for initiating Notch signaling at a high rate.

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The detachment force between Notch receptors and Delta. (A) A force–distance graph generated between an S2-Dl cantilever and an S2-N cell in 1× PBS+Ca 2+. (B) Detachment forces between S2-Dl or S2-N cantilevers and different cells. (C) Force–distance graphs generated when the same S2-Dl cantilever was used successively on an S2-N cell, an S2-NΔ1-18 cell, and an S2-N cell, in 1× PBS+Ca 2+. (D) Cell aggregates of S2-Dl or S2 cells with S2 cells expressing Notch receptors. (E) Correlation (r) between the detachment force and the average number of cells in aggregates (at 10 min). Cell number = aggregation size/S2-NΔ1-18+S2-Dl aggregate size (∼1 cell); SD: N = 20.44, N1-2155 = 8.4, Nnd3 = 13.9, Nmf = 16.7). (F) Aggregated cells after forceful separation. White arrows = Notch cluster regions coextensive with Dl contact regions; black arrow = an N1-2155 cluster region partially coextensive with Dl contact region. Left, Texas red images with an N antibody (C458.2H); right, Nomarski images of the same cells. Bars, 2 μm (in F).
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fig2: The detachment force between Notch receptors and Delta. (A) A force–distance graph generated between an S2-Dl cantilever and an S2-N cell in 1× PBS+Ca 2+. (B) Detachment forces between S2-Dl or S2-N cantilevers and different cells. (C) Force–distance graphs generated when the same S2-Dl cantilever was used successively on an S2-N cell, an S2-NΔ1-18 cell, and an S2-N cell, in 1× PBS+Ca 2+. (D) Cell aggregates of S2-Dl or S2 cells with S2 cells expressing Notch receptors. (E) Correlation (r) between the detachment force and the average number of cells in aggregates (at 10 min). Cell number = aggregation size/S2-NΔ1-18+S2-Dl aggregate size (∼1 cell); SD: N = 20.44, N1-2155 = 8.4, Nnd3 = 13.9, Nmf = 16.7). (F) Aggregated cells after forceful separation. White arrows = Notch cluster regions coextensive with Dl contact regions; black arrow = an N1-2155 cluster region partially coextensive with Dl contact region. Left, Texas red images with an N antibody (C458.2H); right, Nomarski images of the same cells. Bars, 2 μm (in F).

Mentions: A force–distance graph generated between an S2-Dl cantilever and an S2-N cell is shown in Fig. 2 A. The contact force between an S2-Dl cantilever and an S2-N cell was generally ∼19 ± 8 nanoNewtons (nN), and the detachment force ∼14 nN (see also Fig. 2 B, set 1; Fig. 2 C, top graph). These values were very similar in over 25 defined experiments. The contact forces were more variable than the detachment forces, but there was no correlation between the two within each cell line (unpublished data). For measurement of detachment force, we consider only the highest peak, as it is most likely to represent detachment from Dl. The distance of “pull” between S2-N and S2-Dl cells was generally ∼750 nanometers (nm) and could include minor detachment, nonspecific events, stretching of N and/or Dl molecules, or stretching of S2-N and/or S2-Dl cells.


The adhesion force of Notch with Delta and the rate of Notch signaling.

Ahimou F, Mok LP, Bardot B, Wesley C - J. Cell Biol. (2004)

The detachment force between Notch receptors and Delta. (A) A force–distance graph generated between an S2-Dl cantilever and an S2-N cell in 1× PBS+Ca 2+. (B) Detachment forces between S2-Dl or S2-N cantilevers and different cells. (C) Force–distance graphs generated when the same S2-Dl cantilever was used successively on an S2-N cell, an S2-NΔ1-18 cell, and an S2-N cell, in 1× PBS+Ca 2+. (D) Cell aggregates of S2-Dl or S2 cells with S2 cells expressing Notch receptors. (E) Correlation (r) between the detachment force and the average number of cells in aggregates (at 10 min). Cell number = aggregation size/S2-NΔ1-18+S2-Dl aggregate size (∼1 cell); SD: N = 20.44, N1-2155 = 8.4, Nnd3 = 13.9, Nmf = 16.7). (F) Aggregated cells after forceful separation. White arrows = Notch cluster regions coextensive with Dl contact regions; black arrow = an N1-2155 cluster region partially coextensive with Dl contact region. Left, Texas red images with an N antibody (C458.2H); right, Nomarski images of the same cells. Bars, 2 μm (in F).
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fig2: The detachment force between Notch receptors and Delta. (A) A force–distance graph generated between an S2-Dl cantilever and an S2-N cell in 1× PBS+Ca 2+. (B) Detachment forces between S2-Dl or S2-N cantilevers and different cells. (C) Force–distance graphs generated when the same S2-Dl cantilever was used successively on an S2-N cell, an S2-NΔ1-18 cell, and an S2-N cell, in 1× PBS+Ca 2+. (D) Cell aggregates of S2-Dl or S2 cells with S2 cells expressing Notch receptors. (E) Correlation (r) between the detachment force and the average number of cells in aggregates (at 10 min). Cell number = aggregation size/S2-NΔ1-18+S2-Dl aggregate size (∼1 cell); SD: N = 20.44, N1-2155 = 8.4, Nnd3 = 13.9, Nmf = 16.7). (F) Aggregated cells after forceful separation. White arrows = Notch cluster regions coextensive with Dl contact regions; black arrow = an N1-2155 cluster region partially coextensive with Dl contact region. Left, Texas red images with an N antibody (C458.2H); right, Nomarski images of the same cells. Bars, 2 μm (in F).
Mentions: A force–distance graph generated between an S2-Dl cantilever and an S2-N cell is shown in Fig. 2 A. The contact force between an S2-Dl cantilever and an S2-N cell was generally ∼19 ± 8 nanoNewtons (nN), and the detachment force ∼14 nN (see also Fig. 2 B, set 1; Fig. 2 C, top graph). These values were very similar in over 25 defined experiments. The contact forces were more variable than the detachment forces, but there was no correlation between the two within each cell line (unpublished data). For measurement of detachment force, we consider only the highest peak, as it is most likely to represent detachment from Dl. The distance of “pull” between S2-N and S2-Dl cells was generally ∼750 nanometers (nm) and could include minor detachment, nonspecific events, stretching of N and/or Dl molecules, or stretching of S2-N and/or S2-Dl cells.

Bottom Line: Notch signaling is repeatedly used during animal development to specify cell fates.Reduced turnover or Delta pulling accelerate this loss.These data suggest that strong adhesion between Notch and Delta might serve as a booster for initiating Notch signaling at a high rate.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Molecular Genetics, The University of Vermont, VT 05405, USA.

ABSTRACT
Notch signaling is repeatedly used during animal development to specify cell fates. Using atomic force microscopy on live cells, chemical inhibitors, and conventional analyses, we show that the rate of Notch signaling is linked to the adhesion force between cells expressing Notch receptors and Delta ligand. Both the Notch extracellular and intracellular domains are required for the high adhesion force with Delta. This high adhesion force is lost within minutes, primarily due to the action of Presenilin on Notch. Reduced turnover or Delta pulling accelerate this loss. These data suggest that strong adhesion between Notch and Delta might serve as a booster for initiating Notch signaling at a high rate.

Show MeSH
Related in: MedlinePlus