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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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Related in: MedlinePlus

Temporal change in detachment force and SuH/Nintra signaling. (A) Detachment forces between the S2-Dl cantilevers and the S2 cells expressing Notch receptors, in 1× PBS+Ca2+. (B) Western blot showing the levels of N after treatment with S2-Dl cells. (C and D) Western blots showing the levels of active Psn in the different cell mixtures. Asterisk corresponds to a cross-reacting band. (E) Western blots showing the increase in levels of S2- and S3-cleaved N fragments in S2-Dl–treated S2-N cells. (F) Northern blots showing the levels of E(spl)C m3 RNA in the different cell mixtures. (G) Semi-quantitative PCR showing relative levels of E(spl)C m3 RNA. rp49 RNA amplification shows levels of total RNA in the samples.
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fig5: Temporal change in detachment force and SuH/Nintra signaling. (A) Detachment forces between the S2-Dl cantilevers and the S2 cells expressing Notch receptors, in 1× PBS+Ca2+. (B) Western blot showing the levels of N after treatment with S2-Dl cells. (C and D) Western blots showing the levels of active Psn in the different cell mixtures. Asterisk corresponds to a cross-reacting band. (E) Western blots showing the increase in levels of S2- and S3-cleaved N fragments in S2-Dl–treated S2-N cells. (F) Northern blots showing the levels of E(spl)C m3 RNA in the different cell mixtures. (G) Semi-quantitative PCR showing relative levels of E(spl)C m3 RNA. rp49 RNA amplification shows levels of total RNA in the samples.

Mentions: To determine the kinetics of the detachment forces between Dl and the Notch receptors, we rested the S2-Dl cantilevers on S2-N, S2-Nnd3, S2-Nmf, S2-N1-2155, and S2-NΔ1-18 cells for various lengths of time and measured detachment forces. The same batch of S2-Dl cells and the same AFM settings were used on all Notch receptors. Any uncontrolled factors (like drift of the cantilever over time, etc.) were expected to be common to all Notch receptors. Results, shown in Fig. 5 A, indicate the following: (1) the detachment force increased in the first few minutes with all Notch receptors that are able to bind Dl and then decreased for all Notch receptors; (2) the decrease in detachment forces was most rapid with S2-N cells (from 14–18 nN to 0 nN in 10 min), followed by S2-Nnd3 and S2-Nmf cells (from 5–11 nN to 0 nN in 20 min); (3) the detachment force with S2-N1-2155 cells decreased at the slowest rate and did not reach zero even after 60 min (unpublished data); and (4) the adhesion force with S2-NΔ1-18 cell was zero at all times. Thus, there was a positive correlation between the initial detachment force and the rate of decrease in detachment force.


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)

Temporal change in detachment force and SuH/Nintra signaling. (A) Detachment forces between the S2-Dl cantilevers and the S2 cells expressing Notch receptors, in 1× PBS+Ca2+. (B) Western blot showing the levels of N after treatment with S2-Dl cells. (C and D) Western blots showing the levels of active Psn in the different cell mixtures. Asterisk corresponds to a cross-reacting band. (E) Western blots showing the increase in levels of S2- and S3-cleaved N fragments in S2-Dl–treated S2-N cells. (F) Northern blots showing the levels of E(spl)C m3 RNA in the different cell mixtures. (G) Semi-quantitative PCR showing relative levels of E(spl)C m3 RNA. rp49 RNA amplification shows levels of total RNA in the samples.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172611&req=5

fig5: Temporal change in detachment force and SuH/Nintra signaling. (A) Detachment forces between the S2-Dl cantilevers and the S2 cells expressing Notch receptors, in 1× PBS+Ca2+. (B) Western blot showing the levels of N after treatment with S2-Dl cells. (C and D) Western blots showing the levels of active Psn in the different cell mixtures. Asterisk corresponds to a cross-reacting band. (E) Western blots showing the increase in levels of S2- and S3-cleaved N fragments in S2-Dl–treated S2-N cells. (F) Northern blots showing the levels of E(spl)C m3 RNA in the different cell mixtures. (G) Semi-quantitative PCR showing relative levels of E(spl)C m3 RNA. rp49 RNA amplification shows levels of total RNA in the samples.
Mentions: To determine the kinetics of the detachment forces between Dl and the Notch receptors, we rested the S2-Dl cantilevers on S2-N, S2-Nnd3, S2-Nmf, S2-N1-2155, and S2-NΔ1-18 cells for various lengths of time and measured detachment forces. The same batch of S2-Dl cells and the same AFM settings were used on all Notch receptors. Any uncontrolled factors (like drift of the cantilever over time, etc.) were expected to be common to all Notch receptors. Results, shown in Fig. 5 A, indicate the following: (1) the detachment force increased in the first few minutes with all Notch receptors that are able to bind Dl and then decreased for all Notch receptors; (2) the decrease in detachment forces was most rapid with S2-N cells (from 14–18 nN to 0 nN in 10 min), followed by S2-Nnd3 and S2-Nmf cells (from 5–11 nN to 0 nN in 20 min); (3) the detachment force with S2-N1-2155 cells decreased at the slowest rate and did not reach zero even after 60 min (unpublished data); and (4) the adhesion force with S2-NΔ1-18 cell was zero at all times. Thus, there was a positive correlation between the initial detachment force and the rate of decrease in detachment force.

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